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Martynova, E. "HACAT AS A MODEL FOR KERATINOCYTESTRANSFORMATION." Doctoral thesis, Università degli Studi di Milano, 2013. http://hdl.handle.net/2434/221055.
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Mutations of the tumor suppressor gene p53 occur in more than 50% of
human malignancies and lead to the loss of suppressor activity. Moreover
frequently p53 mutants gain novel, oncogenic properties by transcriptional
activation of the genes, involved in cellular proliferation, cell survival and
angiogenesis. Today’s challenge is to understand mechanisms underlying the
gain-of-function of mutant p53 proteins.
By the example of KLF4 promoter we demonstrate that mutant p53 can
carries out its gain-of-function by interaction with another p53 family member,
transcription factor p63. In the first report we provide strong evidence that KLF4
is negatively controlled by p63 in normal skin in the presence of physiological
levels of wild type p53 (wtp53) and that this regulation is subverted by
oncogenic mutations of p53, establishing a direct link between these TFs,
commonly overexpressed in squamous cell carcinoma (SCC).
These results inspired us to investigate the mechanisms of mutant p53
gain-of-function on the genome scale. In the second manuscript we demonstrate
that mutant p53 HaCaT alleles are pro-growth and mutp53 have thousands of
binding sites in the human genome; they affect gene expression profoundly,
both by binding with p63 to consensus elements and by being tethered by other
TFs to their locations.
Although 2 mutant p53 alleles are definitely gain-of-function in the
HaCaT, they are not sufficient to render these cells malignant. Based on the
sphere-forming assay we developed novel model for the tumorigenic conversion
of the HaCaT cells: while immortalized keratinocytes display transformed
phenotype in vitro and not tumorigenic upon injection into nude mice, HaCaTderived
spheres give rise to the SCC in vivo. Thus, this simple model can be
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useful for the future studies of the genetic and phenotypic signatures during
SCC initiation and development.
There is an indication that mutant p53 can execute its gain-of-function
via interaction with NF-Y transcription factor. Ongoing study is devoted to the
investigation of NF-YA subunit role in the processes of cellular proliferation
and apoptosis in the cells with different p53 status.
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Hansen, Kari Ellen. "Utvidet studie av PLA2-uttrykk i HaCaT-keratinocytter :." Thesis, Norges teknisk-naturvitenskapelige universitet, Institutt for biologi, 2011. http://urn.kb.se/resolve?urn=urn:nbn:no:ntnu:diva-13203.
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I denne masteroppgaven har det blitt utviklet primerpar for 17 humane PLA2-isotyper. Ved hjelp av primerene har vi påvist uttrykk av 17 PLA2-isotyper i HaCaT-keratinocytter, hvorav 11 av disse representerer førstegangspåvisninger i HaCaT. Funnene representerer økt kunnskap om PLA2-familien og viser at HaCaT uttrykker et mangfold av PLA2-isotyper. Primerene ble også benyttet til å kartlegge PLA2-uttrykk gjennom keratinocytters differensiering. Resultatene viser at 13 av 17 isotyper uttrykkes gjennom hele differensieringsprosessen, som indikerer at PLA2-enzymer spiller sentrale roller i lipidmetabolisme i human epidermis. PLA2G2A viste, som den eneste av de studerte isotypene, sterk oppregulering under differensieringen, som vitner om at enzymet kan spille en viktig rolle i de øvre strata i epidermis.Sammendrag Under utviklingen av primerparene ble human postnatal placenta tatt i bruk som antatt positiv kontroll for uttrykk av samtlige PLA2-isotyper. 8 av 17 primerpar detekterte mRNA i placenta, hvorav PLA2 gruppe 2D, 4C, 4D og 10 representerer førstegangspåvisninger i human placenta. Sammendrag Primerene utgjør et verktøy som gjør det mulig å kartlegge PLA2-uttrykk i alle humane celler og vev. Da diverse PLA2-isotyper har blitt vist å spille en sentral rolle i inflammasjonssykdommer ved å katalysere hydrolysen av arakidonsyre fra glyserofosfolipider, vil det være intressant å kartlegge PLA2-uttrykk i friskt vev versus betent vev. Primerene vil legge grunnlaget for økt forståelse av de mange PLA2-enzymenes funksjoner i inflammasjonssammenheng, og potensielt legge grunnlag for utvikling av målrettede medisiner mot kronisk inflammatoriske sykdommer.
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Oehme, Martina. "Veränderte Wachstumsfaktorexpression in Spätstadien der Tumorprogression von HaCaT-ras-Zellen /." Heidelberg, 2008. http://opac.nebis.ch/cgi-bin/showAbstract.pl?sys=000259538.
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Goltz, Gerit. "Charakterisierung von Ceramidase-Inhibitoren an der humanen Keratinozyten-Zellinie HaCaT." [S.l.] : [s.n.], 2002. http://www.diss.fu-berlin.de/2002/180/index.html.
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Kors, Christian. "Regulation der Freisetzung von SCF aus proliferierenden versus differenzierenden Keratinozyten/HaCaT." Doctoral thesis, Humboldt-Universität zu Berlin, Medizinische Fakultät - Universitätsklinikum Charité, 2006. http://dx.doi.org/10.18452/15499.
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Der humane Stammzellfaktor (SCF) ist ein zentraler Wachstumsfaktor für Mastzellen in der Dermis und für Melanozyten in den Basalzellschichten der Epidermis. Er wird u. a. von Keratinozyten produziert. In dieser Arbeit wurde die mögliche Regulation der Expression von SCF aus Keratinozyten durch All-Trans-Retinsäure (RA) und Dexamethason in vitro an Hand der HaCaT-Zelllinie untersucht. Die HaCaT-Zellen wurden mit den beiden o. g. Substanzen (10 hoch -5 M bis 10 hoch -9 M) über 24 Stunden und 11 Tage inkubiert. Die Auswertung der HaCaT-Zellzahl, des Gesamt-Proteins SCF, dessen Splicevarianten (mSCF, sSCF) und der Rezeptoren von RA (RAR-alpha, -beta, -gamma) und von Dexamethason (GR-alpha, -beta) erfolgte mittels ELISA und RT-PCR. Dabei ergaben sich folgende Resultate: RA bewirkt einen Anstieg von SCF, Dexamethason bewirkt bei Kurzinkubation eine deutliche Zunahme von SCF, bei Dauerinkubation einen starken Abfall. Die RA-Rezeptoren RA-alpha und -gamma waren nach Inkubation mit RA verstärkt nachzuweisen; die Glukokortikoid-Rezeptoren GR-alpha und -beta zeigten nach Inkubation mit Dexamethason ebenfalls eine vermehrte Expression. Die Expression des Mastzellwachstumsfaktors SCF könnte deshalb unter physiologischen, pathologischen und therapeutischen Bedingungen durch Retinoide und Glukokortikoide reguliert sein.The human stem cell factor (SCF) is a crucial growth factor for mast cells in the dermis and for the melanocytes in the basal layers of the epidermis. SCF is produced, among others, by keratinocytes. This study examines the possible regulation of the expression of SCF from keratinocytes by all-trans retinoic acid (RA) and dexamethasone in vitro by the keratinocyte cell line HaCaT. The HaCaT-cells were incubated for 24 hours or 11 days, respectively, with one of the above mentioned substances (10 to the power of -5 M to 10 to the power of -9 M). The analysis of the number of HaCaT-cells, of the total SCF protein, its splice variants (mSCF, sSCF), the receptors of RA (RAR-alpha, -beta, -gamma), and of the dexamethasone (GR-alpha, -beta) was done by ELISA and RT-PCR. The following results were found: RA induces an increase of SCF, dexamethasone at a short incubation period a considerable increase of SCF, and at long-term incubation a strong decrease. The RA-receptors RA-alpha und -gamma expression is increased after incubation with RA, and the glucocorticoid-receptors GR-alpha and -beta after the incubation with dexamethasone. Therefore, it is probable that the increase of the mast cell growth factor SCF under physiological, pathological and therapeutic conditions could be regulated by retinoic acid and glucocorticoids.
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Elbadawy, Hossein Mostafa. "Studies on the start family of lipid trafficking proteins in HaCaT keratinocytes." Thesis, Glasgow Caledonian University, 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.554307.
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Terminally differentiating keratinocytes actively synthesize and accumulate lipids to maintain the production of lipid lamellae and an effective epidermal barrier. This thesis explored the role of steroidogenic acute regulatory (StAR) related-lipid transfer (START) proteins in the immortalized HaCaT keratinocyte cell line. Differing cell culture conditions were used to study differentiation of HaCaT keratinocytes, including the 'calcium switch' model, studied over a period of 21 days. Gene expression of five 'markers' of keratinocyte differentiation, KRTI, KRTIO, KRTI4, INVand LOR, was used to define progression of this process, revealing a dependency on calcium concentration, and increasing confluency. We then investigated gene expression of the START family of lipid trafficking proteins in HaCaT keratinocytes, compared with primary human keratinocytes. The same nine members of the family, STARDI, STARD2, STARD3, STARD4, STARD5, STARD7, STARDIO, STARD11 and STARDI2 were expressed in both cell types, although levels of STARD2, STARD5 and STARDI2 mRNA proved higher in HaCaT cells than in primary keratinocytes, and gene expression of STARD4 was lower in HaCaT cells compared with primary cells. During HaCaT differentiation, marked changes in gene expression of this family of proteins were noted, reflecting changes in lipid metabolism occurring during this process, the most notable being an increase in cholesterol mass after 21 days in cells exposed to I.4mM calcium compared with cells cultured in calcium depleted media. Steady state levels of mRNA encoding STARDI, D3, D2, D7, DID and Dll tended to increase during keratinocyte differentiation, while STARD4 and STARD5 showed a more complex pattern of expression, with decreased expression observed in the presence of I.4mM calcium. In order to investigate the function of the cytosolic cholesterol binding proteins, STARD4 and STARD5, in HaCaT cells, transient transfections were performed (48h) with pCMV6 vector encoding full length STARD4 or STARD5. Confocal microscopy confirmed the cytosolic location of both proteins, while in 3-D organotypic cultures STARD5 eo-localised with Keratin 10 rather than Loricrin, suggesting a locale in the spinous layer of these cultures. While both cholesterol trafficking proteins repressed cholesterol and cholesteryl ester biosynthesis from [I 4C] acetate, consistent with their proposed role as directional sterol transporters, these changes were associated with distinct gene expression patterns. Overexpression of ST ARD4 was associated with induction of Loricrin mRNA and protein, while STARD5 overexpression repressed gene expression of keratin 1, indicating that altered intracellular cholesterol transport can influence keratinocyte differentiation status. Further, ST ARD4 overexpression was associated with induction of SREBF2 and ABCG4, and repression of ABCAI, while overexpression of STARD5 resulted in induction of PPARD, PPARG and ABCAI, and repression of SREBF2 and LDLR. These findings confirm the impact of these cytosolic sterol transporters on keratinocyte lipid metabolism, and may suggest that ST ARD5 may be more efficient in this regard. In conclusion, manipulation of intracellular lipid transport proteins may therefore provide novel therapeutic strategies for the treatment of lipid-related skin disorders.
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Szkoda, Blake E. "THE EFFECTS OF CITRAL ON CASPASE-3 ACTIVATION IN M624 AND HaCaT CELLS." Marietta College Honors Theses / OhioLINK, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=marhonors1463566418.
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Bretland, Amanda Jane. "Growth, survival and cell death in the epithelial cell lines HaCaT, HT29 and SW742." Thesis, University of Sheffield, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.286914.
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Zanoni, Thalita Boldrin. "Avaliação do perfil de citotoxicidade, mutagenicidade e genotoxicidade dos corantes Basic Red 51, Basic Yellow 57 e P-Fenilenodiamina usados na tintura de cabelo em células da pele." Universidade de São Paulo, 2014. http://www.teses.usp.br/teses/disponiveis/60/60134/tde-31102014-144039/.
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O processo de coloração de cabelos é um dos métodos de tintura mais antigos. No século XIX, iniciou-se a produção de corantes sintéticos, a partir do desenvolvimento da pfenilenodiamina (PPD). Os corantes de cabelo são classificados de acordo com seu mecanismo de ação. Os corantes permanentes são classificados por mecanismos oxidativos, enquanto os corantes diretos colorem a fibra capilar por mecanismos não oxidativos. A investigação sobre os possíveis danos á saúde humana, que podem ser resultantes da exposição de corantes de cabelos, têm sido um tema de enorme desafio para a comunidade cientifica. Particularmente, devido à enorme discrepância dos estudos epidemiológicos e estudos que empregam metodologias in vitro. Neste trabalho, foram investigadas a capacidade citotóxica de um composto representante de cada classe de tinturas de cabelo, um corante temporário (Basic Yellow 57 (BY57), um semi-temporário (Basic Red 51(BR51) e um ingrediente permanente p-fenilinodiamina (PPD) em linhagens de células de pele humana. As linhagens normais da pele humana estudadas foram os queratinócitos imortalizados humanos (HaCaT) e fibroblastos primários, utilizou-se também melanoma SK-Mel-103. Posteriormente, após caracterização do corante mais tóxico, foi investigado o tipo de morte celular, as possíveis alterações destes compostos no ciclo celular, a capacidade de geração de espécies reativas de oxigênio (EROs) e aplicação em cultura tridimensional de pele artificial. Posteriormente, foi avaliada a capacidade de cada corante em induzir estresse oxidativo em queratinócitos humanos (HaCaT), que são a primeira via de exposição de corantes de cabelos. Em seguida, o corante elegido mais tóxico foi aplicado em pele humana provenientes de cirurgia. Finalmente, o potencial de mutagenicidade dos corantes BY57 e BR51 foram avaliados.The process involving hair dyes is one of the oldest methods of coloring. The use of synthetic hair dyes started in the nineteenth century, after the development of p-phenylenodiamine (PPD). The hair dyes are classified according to their mechanism of action. The permanent hair dyes are classified by oxidative mechanisms, while direct dyes color the hair fiber by non-oxidative mechanisms. Research regarding the potential damage of hair dyes to human health has been an enormous challenge for the scientific community. Particularly due to the large discrepancy of epidemiological studies involving in vitro methodologies. In this study, we evaluated the cytotoxic potential of a compound representative from each of the class of hair dyes, a temporary dye (Basic Yellow 57 (BY57), a semi temporary (Basic Red 51 (BR51) and a permanent hair dyes p-phenylenodiamine (PPD) in human skin cells. The studied skin cell lines where, immortalized human keratinocytes (HaCaT) primary fibroblasts, we also used melanoma SK-Mel-103. Subsequently, after characterization of the most toxic dye, we investigated specific mechanisms of cell death, changes in cell cycle and the ability to generate reactive oxygen species (ROS) followed by the evaluation of three-dimensional artificial skin. In addition, we assessed the ability of each dye in inducing oxidative stress in immortalized human keratinocytes (HaCaT) this is the primary route of exposure of hair dyes. Then, the most toxic compound was tested in human skin explants. Finally the mutagenic potential of the dyes BY57 and BR51 were evaluated.
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Raithel, Kerstin. "Effekt von S-Lost in HaCaT-Zellkulturen : Induktion der Apoptose und Effekt eines PARP-Inhibitors /." München, 2008. http://opac.nebis.ch/cgi-bin/showAbstract.pl?sys=000254372.
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Nici, Marco [Verfasser], and Peter [Akademischer Betreuer] Angel. "IL-6 induzierte Signaltransduktion in HaCaT-ras A5 und Fibroblasten / Marco Nici ; Betreuer: Peter Angel." Heidelberg : Universitätsbibliothek Heidelberg, 2013. http://d-nb.info/1177811367/34.
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Stooss, Arnim. "Die Rolle des Transkriptionsfaktors AP-1 bei der Ceramid-induzierten Apoptose in der humanen Keratinozytenzellinie HaCaT." [S.l.] : [s.n.], 1999. http://www.diss.fu-berlin.de/1999/122/index.html.
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Morgene, Mohamed Fedy. "Modélisation in vitro de la colonisation à staphylococcus aureus ; interactions avec l’infection à rhinovirus." Thesis, Lyon, 2018. http://www.theses.fr/2018LYSES054/document.
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Certains virus respiratoires comme rhinovirus semblent favoriser la colonisation par staphylococcus aureus. Cependant, les détails des mécanismes impliqués dans cette synergie n’ont pas été suffisamment élucidés. Le but de cette thèse a été de développer et valider un modèle in vitro mimant la colonisation du vestibule nasal par s. aureus en utilisant les kératinocytes humains hacat. Ce modèle a permis d’étudier (i) les pouvoirs d’adhésion et d’internalisation d’une collection de souche clinique de s. aureus, (ii) l’efficacité intracellulaire des molécules antimicrobiennes utilisées dans le cadre de la décolonisation nasale de s. aureus, (iii) l’effet de la clarithromycine sur l’infection par rhinovirus et (iv) l’impact de l’infection par rhinovirus ou de l’inflammation non spécifique sur la colonisation par s. aureus. ce travail a principalement permis d’identifier un nouveau mécanisme alternatif de l’internalisation de s. aureus à travers la liaison entre la protéine bactérienne eap (extracellular adherence protein) et le récepteur cellulaire icam-1 (intracellular adhesion molecule 1). Cette voie alternative est favorisée en cas d’infection par rhinovirus ou d’inflammation, ce qui pourrait expliquer les observations cliniques de l’augmentation de la charge de s. aureus ou du risque d’infection par cette bactérie lors des infections virales respiratoires ou d’inflammation post-traumatique. Les résultats de cette thèse illustrent la complexité des interactions entre les cellules épithéliales de la muqueuse, s. aureus et les pathogènes viraux et ouvrent les perspectives sur d’autres études nécessaires afin de proposer des stratégies préventives ou thérapeutiques adaptéesSome respiratory viruses such as rhinoviruses seem to promote staphylococcus aureus colonization. However, the details of the bacterial and cellular mechanisms involved in this synergy have not been sufficiently elucidated. The aim of this thesis was to develop and validate an in vitro model mimicking s. aureus colonization of the nasal vestibule by using hacat human keratinocytes. This model allowed to study (i) the adhesion and internalization capacities of various clinical s. aureus strains, (ii) the intracellular efficiency of the antimicrobial molecules used for s. aureus nasal decolonization, (iii) the effect of clarithromycin on rhinovirus infection, and (iv) the impact of rhinovirus infection and non-specific inflammation on s. aureus colonization. This work has mainly identified a new alternative mechanism for the internalization of s. aureus through the binding between the bacterial protein eap (extracellular adherence protein) and the cell receptor icam-1 (intracellular adhesion molecule 1). This alternative pathway is favored in case of rhinovirus infection or inflammation; which could explain the clinical observations of the increase of the load of s. aureus or the risk of infection by this bacterium during respiratory viral infections or post-traumatic inflammations. The results of this thesis illustrate the complexity of the interactions between the mucosal epithelial cells, s. aureus and viral pathogens and suggest that other studies are needed to propose appropriate preventive or therapeutic strategies
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Tonolli, Paulo Newton. "Photosensitization of Lipofuscin in Skin Keratinocytes: Effect of Visible Light on Human Skin." Universidade de São Paulo, 2018. http://www.teses.usp.br/teses/disponiveis/46/46131/tde-12122018-114252/.
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Lipofuscin is an autofluorescent pigment progressively accumulated during cellular aging, in several tissues, such as heart, muscle and retina, especially in the postmitotic period. That phenomenon may result from oxidative stress, when biomolecules and organelles (mainly mitochondria) are damaged, generating non-degradable products inside lysosomes. Lipofuscin can be photosensitized, promoting photoxidative processes in cellular components. Many studies on lipofuscin were made using the human retinal pigment epithelial cells, but very little is known about lipofuscin from human skin. In this work we investigated the photoinduced formation (UVA and visible light) of lipofuscin and the consequence of its photosensitization by visible light. We also established an efficient protocol for the induction of lipofuscinogenesis, through specific damage in mitochondria and lysosomes. Cells that accumulated lipofuscin, after exposure to UVA and blue light, became sensitive to visible light (400-750 nm). We characterized the absorption and fluorescence emission of lipofuscin, as well as its fluorescence lifetime through the time resolved fluorescence microscopy (FLIM). We observed that lipofuscin in keratinocytes has absorption maximum in the blue region of light spectrum (420-450 nm), and maximum emission in the red. When photosensitized at 466 nm, lipofuscinloaded HaCaT cells had reduced cell viability, which was related with singlet oxygen generation, accumulated 8-oxo-dG premutagenic lesions and breaks in the DNA strand. Besides, we investigated the efficiency of different wavelengthsin visible light spectrum (408, 466, 522 and 650 nm) to promote lipofuscin formation due to damages in both mitochondria and lysosomes. Blue (408 and 466 nm) and green light (522 nm), but not red light (650 nm), promoted damage in mitochondria (membrane and DNA integrity) and lysosomes (membrane integrity and autophagic activity), effectively inducing lipofuscinogenesis. Thus, in addition to UVA, visible spectrum itself increases the sensitivity of keratinocytes to the visible light, through the generation of lipofuscin. Finally, we tested the carcinogenic potential of high-energy blue light (408 nm), by chronically irradiating HaCaT cells. For the first time in the literature, the formation of pyrimidine cyclobutane (CPD) dimers in the nuclear DNA of HaCaT cells was observed immediately or after several cycles of irradiation at 408 nm. We identified four major changes involved with the process of malignant transformation: genomic instability, decrease in the expression of tumor suppressor protein p16INK4a, increase in the proliferation rate and resistance to UVA-induced apoptosisA lipofuscina é um pigmento autofluorescente acumulado progressivamente durante o envelhecimento celular em diversos tecidos, como o músculo cardíaco e retina, principalmente no período pós-mitótico. Esse fenômeno pode ocorrer em decorrência do estresse oxidativo, quando biomoléculas e organelas (principalmente mitocôndrias) sofrem danos, gerando produtos não degradáveis no interior dos lisossomos. A lipofuscina pode ser fotossensibilizada promovendo processos fotoxidativos nos componentes celulares. Muitos estudos de lipofuscina foram feitos em células do epitélio pigmentar da retina de olho humano, mas conhece-se muito pouco sobre a lipofuscina de pele humana. Neste trabalho nós investigamos a formação fotoinduzida (UVA e luz visível) de lipofuscina e as consequências da sua fotossensibilização pela luz visível. Nós também estabelecemos protocolos eficazes na indução de lipofuscinogênese, por meio de dano específico em mitocôndrias e lisossomos. Células que acumularam lipofuscina, após exposição à UVA ou luz azul, tornaram-se sensíveis à luz visível (400-750 nm). Caracterizamos as propriedades de absorção e de emissão da lipofuscina e seu tempo de vida de fluorescência, utilizando a microscopia de fluorescência resolvida no tempo (FLIM). Observamos que lipofuscina em queratinócitos tem máximo de absorção na região do azul (420-450 nm), com emissão máxima de fluorescência no vermelho. As células HaCaT carregadas com lipofuscina efotossensibilizadas no visível, tiveram redução da viabilidade celular, que foi relacionada com a geração de oxigênio singlete, bem como acumularam lesões pré-mutagênicas 8-oxo-dG e quebras na fita de DNA. Também, investigamos a eficiência de diferentes comprimentos de onda da luz visível (408, 466, 522 e 650 nm) em promover a formação de lipofuscina em consequência de lesões em mitocôndrias e lisossomos. Tanto a luz azul (408 e 466 nm) quanto a luz verde (522 nm), mas não vermelha (650 nm) promoveram dano em mitocôndrias (integridade de membrana e DNA) e lisossomos (integridade de membrana e atividade autofágica), induzindo eficientemente lipofuscinogênese. Logo, além de UVA, o próprio espectro do visível aumenta a sensibilidade de queratinócitos à luz visível, através da geração de lipofuscina. Por fim, testamos o potencial carcinogênico da luz azul de alta energia (408 nm), irradiando células HaCaT cronicamente. Identificamos quatro mudanças principais envolvidas com o processo de transformação maligna: instabilidade genômica, redução da expressão de proteína supressora de tumor p16INK4a, aumento da taxa de proliferação, e resistência à apoptose. Além disso, a formação de dímeros de pirimidina ciclobutano (CPD) no DNA nuclear de células HaCaT logo após ou depois de vários ciclos de irradiação com 408 nm foi observada pela primeira vez na literatura.
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Wagner, Kathrin. "Überprüfung der Expression des Proopiomelanocortin (POMC)-Gens in HaCaT- und A-431-Zellkulturen unter Basal- und Stimulationsbedingungen." Wettenberg : VVB Laufersweiler, 2005. http://deposit.ddb.de/cgi-bin/dokserv?idn=976093316.
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Betz, Susanne. "Vergleich der Isoprenalin-stimulierten cAMP-Produktion von HaCaT-Keratinozyten und kultivierten Keratinozyten aus gesunder und psoriatischer Haut." [S.l.] : [s.n.], 2002. http://deposit.ddb.de/cgi-bin/dokserv?idn=969112807.
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Hertting, Tatjana. "Einfluss von Tocopherol auf UV-induzierte NF-kB-Regulation [NF-kappaB-Regulation] und Lipidperoxidation in HaCaT-Keratinozyten." [S.l.] : [s.n.], 2004. http://www.diss.fu-berlin.de/2004/300/index.html.
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Bauer, Tanja. "Struktur-Wirkungsbeziehung von Ceramidanaloga in der Regulation von Proliferation, Differenzierung und Apoptose in der immortalisierten humanen Keratinozytenzelllinie HaCaT /." Berlin : Mensch-und-Buch-Verl, 2001. http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&doc_number=009571630&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA.
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Schwarz, Erik [Verfasser]. "Zellbiologischer Einfluss von Schmelzmatrixproteinen auf die Expression von antimikrobiellen Peptiden und Zytokinen in humanen Epithelzellen (HaCaT) / Erik Schwarz." Berlin : Medizinische Fakultät Charité - Universitätsmedizin Berlin, 2016. http://d-nb.info/1121007740/34.
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Marrero, Bernadette. "Evaluation of Immunogene Therapy Using a Plasmid Encoding IL-15 Delivered by Electroporation in a 3D Tumor Model and a Mouse Melanoma Model." Scholar Commons, 2010. http://scholarcommons.usf.edu/etd/3520.
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Melanoma is an aggressive disease with few effective treatment options. Non-toxic, anti-tumor therapies and prophylactic approaches are currently being investigated to identify treatment options that will control and remove late-stage melanoma.
The overall goal of this project was to establish an effective delivery method for a plasmid encoding human interleukin (phIL-15) into mouse melanoma cells (B16.F10) using the gene transfer technique electroporation (EP)1. The EP delivery phIL-15 was optimized using an in vitro 3D tumor model. The purpose was to translate these IL-15 delivery conditions into an in vivo mouse melanoma model to study IL-15 signal transduction and stimulate immune cells to destroy tumor antigens as well as promote an anti-tumor immune memory response.
The in vitro 3D tumor model and the mouse model demonstrated similar expression patterns when delivering phIL-15 with different EP conditions. Intra-tumoral delivery using 500V/cm 20ms enhanced gene delivery and increased IL-15 protein expression compared to 1300V/cm 100μs. There was also a visible increase in transfection efficacy between tumor cells compared to skin cells when delivering pmIL-12 and phIL-15 plasmid constructs in vivo. The plasmid+EP groups 1300V/cm and 500V/cm stimulated increased expression of cytokines IL-1β, IL-6, INFγ, MIP-1β and TNFα. These EP groups also promoted tumor regression by up-regulating CD8+ T cells and CD4+ T cells which targeted melanoma. Regression and survival studies demonstrated that 73.3% of mice cleared B16.F10 cells when treated with phIL-xi15+1300V/cm and pVax+500V/cm. In addition, 53% of the mice responded to the phIL-15+500V/cm treatment group. Furthermore, 75% of the mice from group phIL-15+500V/cm survived secondary inoculation and tumor challenge. In conclusion, plasmid with encoding gene insert phIL-15 delivered by EP has the potential to act as an anti-tumor therapy because it promotes melanoma regression and enhances mouse survival through innate and adaptive cell-mediated immune responses.
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Henri, Pauline. "RECEPTEURS CUTANES A LA MELANOCORTINE DE TYPE 1 (MC1R) ET REPONSES OXYDATIVES AUX UVA DANS DES KERATINOCYTES HUMAINS HaCaT." Phd thesis, Université Montpellier I, 2010. http://tel.archives-ouvertes.fr/tel-00560959.
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Les ultraviolets A (UVA) sont carcinogènes et produisent des espèces réactives de l'oxygène (ERO). Le récepteur à la mélanocortine de type 1 (MC1R) est un récepteur couplé aux protéines G (RCPG) qui est impliqué dans la mélanogénèse et dans l'inflammation cutanée. Certains variants du gène sont associés à un risque accru de mélanomes et de carcinomes cutanés. Le MC1R est exprimé surtout dans les mélanocytes mais son expression peut être induite par les UV in vitro dans les kératinocytes et in vivo dans la peau. Le récepteur MC1R est activé par l'α-MSH. L'objectif de ce travail de thèse a été d'étudier les effets du récepteur MC1R sur le stress oxydatif induit par les UVA dans des lignées kératinocytaires humaines HaCaT exprimant le récepteur MC1R ou son variant non fonctionnel Arg151Cys. Nous avons montré que la production d'ERO intracellulaire induite par les UVA est fortement inhibée dans les cellules HaCaT-MC1R et que cette inhibition est renforcée en présence d'α-MSH. L'inhibition du stress oxydatif induit par les UVA dans les cellules transfectées par le MC1R est en partie dépendante de la phosphorylation de la sous-unité activatrice, NoxA1 de la NADPH oxydase. Le traitement des cellules HaCaT-MC1R par un inhibiteur du récepteur au facteur de croissance épidermique (EGFR) restaure l'habilité de ces cellules à induire un stress oxydatif après irradiation UVA. Ces résultats montrent que l'activité constitutive du récepteur MC1R dans des kératinocytes pourrait inhiber le stress oxydatif induit par les UVA via des mécanismes dépendants de l'AMPc et de l'EGFR.
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Wagner, Kathrin [Verfasser]. "Überprüfung der Expression des Proopiomelanocortin (POMC)-Gens in HaCaT- und A-431-Zellkulturen unter Basal- und Stimulationsbedingungen / Kathrin Wagner." Wettenberg : VVB Laufersweiler, 2005. http://d-nb.info/976093316/34.
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Hausmann, Dominikus [Verfasser], and Martin [Akademischer Betreuer] Leverkus. "Die Bedeutung von cFLIPlong für die Todesrezeptor-abhängige Regulation der Apoptose in HaCaT-Keratinozyten / Dominikus Hausmann. Betreuer: Martin Leverkus." Würzburg : Universitätsbibliothek der Universität Würzburg, 2013. http://d-nb.info/1031630880/34.
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Burga-Sánchez, Jonny 1974. "Efeitos da articaína associada a 2-hidroxipropil-beta-ciclodextrina ou epinefrina sobre a viabilidade celular de queratinócitos humanos (HaCaT)." [s.n.], 2014. http://repositorio.unicamp.br/jspui/handle/REPOSIP/288960.
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Orientadores: Francisco Carlos Groppo, Maria Cristina VolpatoDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: A associação a carreadores tem sido proposta visando prolongar o efeito anestésico, além de reduzir a toxicidade de vários anestésicos locais, incluindo a articaína (ATC). O objetivo do estudo foi avaliar in vitro o efeito da ATC livre ou associada a diferentes concentrações de epinefrina ou 2-hidroxipropil-?-ciclodextrina (HP-?-CD) sobre a viabilidade celular de queratinócitos humanos imortalizados (HaCaT). Foi avaliado também o efeito do metabissulfito de sódio, principal antioxidante e componente das soluções anestésicas comerciais, sobre a viabilidade das citadas células. A microscopia eletrônica de varredura (MEV) foi utilizada para avaliar as características físicas dos cristais da ATC, da HP-?-CD e do complexo de inclusão liofilizado de ATC/HP-?-CD. As células HaCaT foram expostas às formulações de ATC em diferentes concentrações desde 0.1% até 4%, associadas ou não a epinefrina 1:50.000, 1:100.000 e 1:200.000, ou em associação com HP-?-CD; em tempos de 10, 30, 60 e 240 min. As células viáveis foram quantificadas pelo método do MTT após os períodos de exposição e comparadas a um grupo controle sem tratamento. A avaliação celular qualitativa foi realizada por microscopia de fluorescência pelo método de coloração Live/Dead®. A análise estatística foi realizada por two-way ANOVA (teste de Tukey, p>0.05). Os resultados revelaram que a toxicidade da ATC depende da concentração e do tempo de exposição, sendo que quando complexada com HP-?-CD ou associada à epinefrina 1:200.000, houve tendência a diminuir a toxicidade avaliada inicialmente. Da mesma forma, os adjuvantes como a epinefrina, o metabissulfito de sódio e a HP-?-CD sozinhos mostraram biocompatibilidade nas concentrações empregadas neste estudo. Concluímos que a associação da ATC com a HP-?-CD bem como à epinefrina 1:200.000 diminuiu a toxicidade do anestésico local quando avaliado nas concentrações mais baixas. Entretanto, a associação destes adjuvantes não melhorou o perfil de toxicidade da ATC quando avaliado em concentrações clínicas usuais de 2 e 4%
Abstract: The association with carriers has been proposed to prolong the anesthetic effect and reduce the toxicity of several local anesthetics including articaine (ATC). The aim of this study was to assess the in vitro effect of ATC associated with different concentrations of epinephrine or 2-hydroxypropyl-?-cyclodextrin (HP-?-CD) on cell viability in immortalized human keratinocyte cells cultures (HaCaT). It was also evaluated the effect of sodium metabisulphite, major antioxidant component of commercial anesthetic solutions, on the viability of cited cells. The scanning electron microscopy (SEM) was used to assess the physical characteristics of ATC crystals, HP-?-CD and ATC/HP-?-CD lyophilized inclusion complex. The HaCaT cells were exposed to different formulations of ATC in concentrations from 0.1% to 4%, associated or not with epinephrine 1:50.000, 1:100.000 and 1:200.000, or in formulation with HP-?-CD; in 10, 30, 60 and 240 min time exposure. Vital HaCaT cells were quantified by the MTT method after exposure periods and compared to an untreated control group. Cells were assessed qualitatively by fluorescent microscopy using the staining Live/Dead® method. Statistical analysis was performed by two-way ANOVA (Tukey test, p> 0.05). The results showed that toxicity of ATC depends on the concentration and exposure time, and when complexed with HP-?-CD or associated with epinephrine 1:200.000, there was a tendency to decrease the toxicity initially evaluated. Likewise, adjuvants such as epinephrine, sodium metabisulphite and HP-?-CD alone showed biocompatibility in concentrations used in this study. In conclusion, the association of ATC with HP-?-CD as well as epinephrine 1:200.000 decreased local anesthetic toxicity when lower concentrations of ATC are used. However, the combination of adjuvants did not improve the toxicity profile of ATC when used in clinical usual concentrations of 2 and 4%
Mestrado
Farmacologia, Anestesiologia e Terapeutica
Mestre em Odontologia
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Henri, Pauline. "Récepteurs cutanés à la mélanocortine de type 1 (MC1R) et réponses oxydatives aux UVA dans des kératinocytes humains HaCaT." Thesis, Montpellier 1, 2010. http://www.theses.fr/2010MON13520/document.
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Les ultraviolets A (UVA) sont carcinogènes et produisent des espèces réactives de l'oxygène (ERO). Le récepteur à la mélanocortine de type 1 (MC1R) est un récepteur couplé aux protéines G (RCPG) qui est impliqué dans la mélanogénèse et dans l'inflammation cutanée. Certains variants du gène sont associés à un risque accru de mélanomes et de carcinomes cutanés. Le MC1R est exprimé surtout dans les mélanocytes mais son expression peut être induite par les UV in vitro dans les kératinocytes et in vivo dans la peau. Le récepteur MC1R est activé par l'α-MSH. L'objectif de ce travail de thèse a été d'étudier les effets du récepteur MC1R sur le stress oxydatif induit par les UVA dans des lignées kératinocytaires humaines HaCaT exprimant le récepteur MC1R ou son variant non fonctionnel Arg151Cys. Nous avons montré que la production d'ERO intracellulaire induite par les UVA est fortement inhibée dans les cellules HaCaT-MC1R et que cette inhibition est renforcée en présence d'α-MSH. L'inhibition du stress oxydatif induit par les UVA dans les cellules transfectées par le MC1R est en partie dépendante de la phosphorylation de la sous-unité activatrice, NoxA1 de la NADPH oxydase. Le traitement des cellules HaCaT-MC1R par un inhibiteur du récepteur au facteur de croissance épidermique (EGFR) restaure l'habilité de ces cellules à induire un stress oxydatif après irradiation UVA. Ces résultats montrent que l'activité constitutive du récepteur MC1R dans des kératinocytes pourrait inhiber le stress oxydatif induit par les UVA via des mécanismes dépendants de l'AMPc et de l'EGFRUltraviolet A (UVA) radiations are responsible for deleterious effects, mainly due to reactive oxygen species (ROS) production. Alpha-melanocyte stimulating hormone (α-MSH) binds to Melanocortin-1 Receptor (MC1R) in melanocytes to stimulate pigmentation and modulate cutaneous inflammatory responses. MC1R may be induced in keratinocytes after UV exposure. To investigate the effect of MC1R signaling on UVA-induced ROS (UVA-ROS) production, we generated HaCaT cells that stably express human MC1R (HaCaT-MC1R) or the Arg151Cys (R151C) non- functional variant (HaCaT-R151C). We then assessed ROS production immediately after UVA exposure and found that: (1) UVA-ROS production was strongly reduced in HaCaT-MC1R but not in HaCaT-R151C cells compared to parental HaCaT cells; (2) this inhibitory effect was further amplified by α-MSH treatment of HaCaT-MC1R cells before UVA exposure; (3) after UVA irradiation, NoxA1 phosphorylation was increased i n HaCaT-MC1R compared to HaCaT and HaCaT-R151C cells. Inhibition of PKA in HaCaT-MC1R cells resulted in a marked increase of UVA-ROS production; (4) the ability of HaCaT-MC1R cells to produce UVA-ROS was restored by inhibiting epidermal growth factor receptor (EGFR) or extracellular signal-regulated kinases (ERK) activity before UVA exposure. Our findings suggest that constitutive activity of MC1R in keratinocytes may reduce UVA-induced oxidative stress via EGFR and cAMP-dependent mechanisms
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Rekus, Martin T. "Characterization of growth and differentiation of a spontaneously immortalized keratinocyte cell line (HaCaT) in a defined, serum free culture system." [S.l.] : [s.n.], 2000. http://deposit.ddb.de/cgi-bin/dokserv?idn=963505262.
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Heu, Céline. "Vieillissement des barrières biologiques. : caractérisation morphologique et fonctionnelle d'un modèle de stress environnemntal induit chez la lignée kératinocytaire humaine HaCat." Thesis, Besançon, 2012. http://www.theses.fr/2012BESA0005.
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II existe de nombreuses études mettant en relation le vieillissement et le stress oxydant, mais peu d'entre elles ont caractérisé l'évolution des marqueurs des défenses antioxydantes, notamment au niveau cutané, au cours du vieillissement dit extrinsèque ou environnemental.Nous avons cherché à mimer in vitro le vieillissement extrinsèque cutané, à partir d'un modèle de cellules épidermiques en culture, la lignée de kératinocytes humains HaCaT, soumises à un stress chimique, l'exposition au glyphosate, un principe actif entrant dans la composition de nombreuses formulations pesticides. Ainsi, nous avons exploré notre modèle par une approche multi-échelle originale et innovante: tout d'abord, à l'échelle moléculaire, par une étude protéomique quantitative différentielle des taux d'expression protéique intracytosolique suite à l'induction du stress ; puis, à l'échelle cellulaire en cytomètrie en flux, par la caractérisation fonctionnelle du stress oxydant et de la mort cellulaire qui en découle ; enfin, à l'échelle micro- et nanométrique, en microscopie confocale et à force atomique, par le suivi de l'évolution des propriétés morphologiques et viscoélastiques des kératinocytes stressés.Ce travail de thèse a démontré que le glyphosate altérait signifïcativement l'état et la fonction barrière de l'épiderme humain, ciblant notamment les kératinocytes, dont l'équipement moléculaire et les fonctions vitales d'adhérence et de dynamique membranaire se retrouvent fondamentalement dégradées. L'ensemble de ces résultats constitue un « tableau » qui évoque parfaitement les événements, les signaux et les comportements cellulaires s'installant au cours du vieillissement cutanéNumerous studies relate ageing to oxidative stress. Few of them described thé évolution of markers of antioxidant défenses, in particular at thé cutaneous level, during extrinsic or environmental ageing. We tried to mimic in vitro cutaneous extrinsic ageing, with a model of epidermic cells in culture, thé human kératinocytes cell line HaCaT, subjected to a chemical stress, Glyphosate, an active ingrédient présent in thé composition of numerous pesticide formulations. Indeed, we examined thé effects of induced ageing in thé loss of kératinocytes integrity in culture, by an original and innovative multi-scale approach: Firstly, at thé molecular scale, we assessed thé stress induced modifications of intracytosolic protein expression by a differential quantitative proteomic study; then, at thé .cellular scale, with flow cytometry, by thé functional characterization of thé oxidative stress and resulting cell death; fmally, at thé micro- and nanoscale, with confocal and atomic force microscopy by thé following thé évolution of morphological and viscoelastic properties of stressed kératinocytes. This PhD work demonstrated that glyphosate impaired thé state and thé barrier function of thé human skin, in particular by fundamentally impairing kératinocytes through thé molecular equipment, thé vital adhésion fonctions and membrane dynamics. Ail thèse results give us a global image of events, signais and cell behavior occurring in skin aging
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Dykstra, Stefanie Nolte. "INVESTIGAÇÃO DO EFEITO DO TRITERPENO 3Β,6Β,16Β-TRIHIDROXILUP-20(29)-ENO (TTHL) SOBRE A PROLIFERAÇÃO DE QUERATINÓCITOS HUMANOS (HACAT)." UNIVERSIDADE ESTADUAL DE PONTA GROSSA, 2013. http://tede2.uepg.br/jspui/handle/prefix/108.
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Previous issue date: 2013-02-21Introduction: Psoriasis is a skin inflammatory disease characterized by keratinocytes hyperproliferation and their incomplete differentiation in the epidermis. Treatment aims to reduce the psoriatic plaques formation; however, many therapeutic regimens do not produce satisfactory results. Searching for new alternatives, this study evaluated the possible effect of the TTHL compound, isolated from the ethanolic extract of Combretum leprosum, on HaCaT proliferation.Methods: The cellular viability was assessed 24 hours after the treatment with different concentrations of TTHL through MTT and neutral red methods. To evaluate the TTHL anti-proliferative activity, the cells were treated with different concentrations of TTHL, in alternate days (96 hours) and analyzed through MTT and Cyquant methods. The cell cycle was evaluated by flow citometry. The possible toxicity by the topical application of the TTHL (0.3 mg/ear) was assessed in vivo, through skin atrophy, by measuring ear thickness of animals and histology.Results: The MTT assay showed, after 24 hours, that TTHL presents cytotoxic activity in HaCaT cells, at the concentrations of 5, 7, 9 and 10 μM, decreasing, respectively, 53.54, 89.53, 89.69 and 90.30 ± 5.84% the cellular viability compared to the control group. The neutral red assay, in the same concentrations, decreased 39.83%, 48.67%, 47.78% and 50.44 ± 4.42%, respectively. In the cellular proliferation assay, the 5, 7 and 10 μM concentrations decreased 24.27%, 59.25% and 82.30% ± 5.96% the cells number, as showed in the MTT method, and 30.30%, 59.59% e 90.90% ± 6.03% as showed in the Cyquant method. The cell cycle assay showed that, at the concentrations of 5, 7 and 10 μM, the non-viable cells number was higher and statistically significant when compared to the control group. Through the skin atrophy assay was possible to conclude that TTHL does not cause atrophy.Conclusion: TTHL is capable of reducing cell viability. These data suggest that TTHL might be a possible tool for the treatment of hyperproliferative diseases, among them, psoriasis. However, additional studies are needed to verify the action mechanism and triterpene safety.
Introdução: A psoríase é uma doença inflamatória cutânea caracterizada pela hiperproliferação e diferenciação incompleta de queratinócitos na epiderme. O tratamento tem como objetivo reduzir a formação de placas psoriáticas, porém muitos regimes terapêuticos não produzem resultados satisfatórios. Buscando alternativas, este trabalho avaliou o possível efeito do composto TTHL, isolado do extrato etanólico de Combretum leprosum, sobre a proliferação de HaCaT. Métodos: A viabilidade celular foi avaliada 24 horas após o tratamento com diferentes concentrações de TTHL, através do método de MTT e vermelho neutro. Para avaliar a atividade antiproliferativa, as células foram tratadas com diferentes concentrações de TTHL, em dias alternados e analisadas através do método de MTT e Cyquant. O ciclo celular foi avaliado por citometria de fluxo. A possível toxicidade pela aplicação tópica do TTHL foi avaliada in vivo, pelo ensaio de atrofia cutânea, através da medida da espessura da orelha dos animais e histologia. Resultados: No ensaio de MTT, após 24 horas, o TTHL, apresentou atividade citotóxica nas células HaCaT, nas concentrações de 5, 7, 9 e 10 μM, diminuindo, respectivamente em 53,54, 89,53, 89,69 e 90,30 ± 5,84% a viabilidade celular quando comparado ao grupo controle. No ensaio de vermelho neutro, essas mesmas concentrações diminuíram 39,83%, 48,67%, 47,78% e 50,44 ± 4,42%, respectivamente. No ensaio de proliferação celular, nas concentrações de 5, 7 e 10 μM, observou-se diminuição de 24,27%, 59,25% e 82,30% ± 5,96% no número de células, conforme o método de MTT e 30,30%, 59,59% e 90,90% ± 6,03% conforme o método de Cyquant. No ensaio de ciclo celular observou-se que nas concentrações de 5, 7 e 10 μM o número de células não viáveis foi maior e estatisticamente significante quando comparado com o grupo controle. Através do ensaio de atrofia cutânea foi possível concluir que o TTHL não causa atrofia. Conclusão: O TTHL é capaz de diminuir a viabilidade celular. Esses dados sugerem que o TTHL possa ser uma possível ferramenta para o tratamento de doenças hiperproliferativas, dentre elas, a psoríase. Entretanto, estudos adicionais são necessários para verificar o mecanismo de ação e segurança do triterpeno.
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Zghebi, Salwa S. "Crystal engineering, Bio Pharmaceutics and Cell biology of active pharmaceutical ingredient (drug) nanoparticles. Formation and cell interaction of hydrocortisone and prednisolone nanoparticles." Thesis, University of Bradford, 2010. http://hdl.handle.net/10454/5183.
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Nanotechnology applications have emerged enormously in recent times. Of particular interest is that area that overlaps the areas of nanotechnology, biology and medicine: nanomedicine. One advantage of nanomedicines is it that it can be used as an enabling technology by pharmaceutical researchers and industry to overcome issues associated with the low bioavailability of hydrophobic drugs. In the first part of the current study, nanosuspensions of two of hydrophobic steroid drugs: hydrocortisone and prednisolone were produced. Nanosuspensions were prepared using a bottom-up approach: the anti-solvent precipitation method using microfluidic reactors. Surface modification was carried out on these nanosuspensions using cationic surfactants to obtain nanoparticles with different levels of surface positive charge as indicated by ¿-potential values. Dynamic light scattering (DLS) and transmission electron microscope (TEM) techniques were used to characterize the prepared nanoparticles. Powder X-ray diffraction (PXRD) and differential scanning calorimetry (DSC) were also used to characterize hydrocortisone nanoparticles. In the second part, cellular uptake of both coated and uncoated nanoparticles by HaCaT keratinocytes cell line was examined and indicated by quantifying the anti- inflammatory effect of nanoparticles on the LPS-induced inflammation. Also, TEM was employed to evaluate the cellular uptake of hydrocortisone nanoparticles. Results showed higher ant-inflammatory effect of coated nanoparticles over uncoated nanoparticles. Furthermore, the anti-inflammatory effect of coated nanoparticles was correlated to the degree of positive surface charge.Libyan government
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Muniz, Bruno Vilela 1988. "Efeitos da articaína livre e associada a lipossomas com gradiente de pH transmembranar sobre a viabilidade celular e expressão de IL-6 em queratinócitos humanos (HaCaT) : The effects of plain and liposome-associated articaine with transmembrane pH gradient on humam keratinocytes (HaCaT) viability and IL-6 expression." [s.n.], 2014. http://repositorio.unicamp.br/jspui/handle/REPOSIP/287997.
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Orientadores: Maria Cristina Volpato, Francisco Carlos GroppoDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: A articaína não encapsula em proporção significativa em lipossomas sem gradiente de pH transmembranar. O objetivo deste estudo foi preparar e realizar a caracterização inicial de formulação de articaína associada a lipossomas unilamelares (400nm) com gradiente de pH transmembranar, com sulfato de amônia como tampão interno sobre a viabilidade celular em culturas de queratinócitos humanos (HaCaT) e sobre a liberação de uma interleucina pró-inflamatória (IL-6), comparando com formulações de articaína livre. As células foram expostas às formulações de articaína nas concentrações 0,1%, 0,2% e 0,3% na forma de solução e em suspensão lipossomal (lipossomas unilamelares), além dos controles (soro fisiológico, suspensão lipossomal e meio de cultura). A avaliação da viabilidade celular (redução do MTT - espectrofotometria) foi realizada após 10 min e 4h e a quantificação da IL-6 (imunoensaio de ELISA) após 4h da exposição às formulações. Os resultados foram submetidos aos testes de Kruskal-Wallis com post-hoc de Dunn (viabilidade celular) e de Student-Newman-Keuls (IL-6) com significância de 5%. As vesículas lipossomais mantiveram-se integras após a encapsulação de articaína, apresentando 18,95% de eficiência de encapsulação. A polidispersão dos lipossomas sem anestésico foi de 0,2 ± 0,0, enquanto a dos liposomas contendo articaína variou de 0,56 ± 0,03 a 0,66 ± 0,10. O potencial Zeta variou de -10,5 ± 0,9 a -21,2 ± 0,9 mV. O tamanho das vesículas variou de 622 ± 71,5 a 796 ± 111,95 nm. A viabilidade celular foi diminuída após 10 min de exposição às formulações lipossomais (com e sem articaína) em relação aos demais tratamentos (p<0,05); as formulações lipossomais não diferiram entre si (p>0,05). Não houve diferenças entre os demais tratamentos (p>0,05), os quais não alteraram a viabilidade nesse tempo de exposição. Após exposição por 4h houve diminuição na viabilidade em todas as formulações lipossomais e nas formulações de articaína livre 0,2% e 0,3% (p<0,05), à exceção dos grupos controle e articaína 0,1% (p>0,05). A liberação de IL-6 não foi afetada pelas formulações lipossomais (p>0,05); a articaína livre em todas as concentrações testadas aumentou a liberação de IL-6, tanto em relação ao controle, quanto às formulações lipossomais com a mesma concentração de articaína (p<0,05). Conclui-se que a utilização de lipossomas com gradiente de pH transmembranar aumentou a encapsulação de articaína, entretanto, apresentou toxicidade intrínseca no modelo avaliado
Abstract: Articaine is not encapsulated in significant proportion in liposomes without transmembrane pH gradient. The aim of this study was to perform the initial characterization of a formulation of articaine in unilamelar liposome with transmembranarpH gradient and to observe its effects on human keratinocytes (HaCaT) regarding cellular viability and liberation of interleukin 6 (IL-6) in comparison to plain articaine. HaCaT cells were exposed to plain articaine solutions and liposomal suspensions (0.1%, 0.2% and 0.3% articaine concentrations) and to the controls (saline, liposomal suspension and culture medium). Cell viability (MTT reduction - spectrophotometry) was evaluated at 10 min and 4h after exposure to the treatments; IL-6 was determined after 4 h of cell treatments. Cell viability results were submitted to Kruskal-Wallis test, followed by Dunn post-hoc test; IL-6 results were analyzed by Kruskal-Wallis and Student-Newman-Keuls tests. Significance was set at 5%. Liposome vesicles remained intact after articaine loading and presented encapsulation efficiency of 18.95%. Liposome without anesthetic presented polydispersity index of 0.2 ± 0.0, while liposomes with articaine showed values of 0.56 ± 0.03 to 0.66 ± 0.10. Zeta potentials varied from -10.5 ±0.9 to -21.2 ± 0.9 mV and vesicle sizes from 622 ± 71.5 to 796 ± 111.95 nm. Cell viability decreased after 10 min exposure to the liposomal formulations (with and without articaine) in relation to the other treatments (p<0.05); liposomal formulations did not differ from each other (p>0.05). No differences were found among the other treatments (p>0.05), which did not interfere in cell viability after this exposure time. After 4h exposure cell viability was diminished by all liposomal formulations and by 0.2% and 0.3% plain articaine (p<0.05), except for control groups and 0.1% plain articaine (p>0.05). IL-6 release was not affected by liposomal formulations (p>0.05); all concentrations of plain articaine increased IL-6 release in relation to the controls and to each correspondent liposomal formulation (p<0.05). In conclusion, liposome with transmembrane pH gradient increases articaine encapsulation in relation to that described in the literature, however it presented intrinsic in the model evaluated
Mestrado
Farmacologia, Anestesiologia e Terapeutica
Mestre em Odontologia
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Eriksson, Jennifer. "Generation of mutated expression plasmid KRT1 and comparison of HaCaT cells transfected with expression plasmid KRT1 or KRT10 concerning keratin aggregates." Thesis, Uppsala universitet, Institutionen för medicinsk biokemi och mikrobiologi, 2012. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-176426.
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Introduction The genetic skin disease epidermolytic ichtyosis is caused by mutations in either keratin gene 1 or 10 and leads to blisters and hyperkeratosis of the epidermis. At cellular level the disease is seen as aggregates in the keratin filaments. Since medicines are hard to investigate and produce mainly due to lack of reproducible model systems, there is no good treatment available for this disease today. In this article we describe how an in vitro model consisting of cells from a stable cell line transfected with expression plasmids to mimic patient cells, may be a possible alternative for screening compounds for therapies. The first step was to generate an expression plasmid required to complete the in vitro model. The model was tested by a preliminary experiment with 4-phenylbuturate (4-PBA) to see if the substance had an effect on the amount of cells with keratin aggregates. Methods PCR and primers containing the desired mutation were used to incorporate a deletion in wild type keratin gene1 plasmid to generate the expression plasmid. HaCaT cells were transfected with the plasmid for expression of keratin. The percentage of cells with keratin aggregates was assessed by fluorescence microscopy. Results/Conclusion Cells containing mutated plasmid had a higher percentage of keratin aggregates compared to cells transfected with wild type plasmid. 4-PBA was found not to affect the amount of cells with keratin aggregates. According to this project the model might be a useful tool for screening compounds, but it needs to be more developed and tested.
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Oliveira, Gisele Augusto Rodrigues de. "Avaliação toxicogenética e ecotoxicológica de corantes têxteis." Universidade de São Paulo, 2013. http://www.teses.usp.br/teses/disponiveis/60/60134/tde-05092013-152705/.
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O tingimento de tecidos começou há milhares de anos e a disponibilidade comercial de corantes é enorme e crescente. A indústria têxtil brasileira desempenha um papel de inquestionável importância, destacando-se entre as principais atividades econômicas do país. O processo de tingimento é um dos fatores fundamentais no sucesso comercial dos produtos têxteis, uma vez que o consumidor exige cores resistentes à exposição ao calor, à luz, à transpiração e às lavagens. Segundo a literatura, condições de transpiração intensa contribuem para uma alta taxa de migração e subseqüente penetração de corantes têxteis para a pele humana. Além disso, 2 a 50% desses compostos permanecem no banho de tingimento e são descartados nos efluentes industriais, contaminando o ambiente e colocando em risco a saúde humana, uma vez que os métodos convencionais de tratamento de efluentes são ineficientes na remoção da coloração e da mutagenicidade de alguns corantes. Dentro deste contexto, este trabalho teve como objetivo avaliar os efeitos toxicogenéticos do corante Direct Black (DB38) original e após extração por lixiviação com suor sintético, utilizando o teste do cometa com fibroblastos e queratinócitos de pele humana, o teste Anexina V com fibroblastos e o ensaio de mutagenicidade com Salmonella typhimurium. Adicionalmente, foi investigada a ecotoxicidade dos corantes têxteis Direct Black 38 e Reactive Blue 15 (RB15) originais por meio de ensaios com sementes, dapnhias, minhocas e zebrafish realizados na UTOX, em Barcelona. O corante DB38 original e lixiviado não induziram genotoxicidade em fibroblastos e queratinócitos de pele humana. O corante DB38 original foi mutagênico para as linhagens TA98 e TA100 de S. typhimurium na presença de S9. Entretanto, o corante lixiviado não induziu mutagenicidade para essas linhagens testadas, considerando que a maior taxa de migração do corante para a solução de suor foi de ~1% nas seguintes condições: tingimento sem ensaboamento, pH 8,0 e 8 horas de incubação à 42°C. O corante original é citotóxico para fibroblastos após 48 horas de exposição. No entanto, essa citotoxicidade não foi mais observada após a lixiviação no suor. Os corantes DB38 e RB15 originais não foram tóxicos para as sementes de pepino, alface e tomate, e nem para as minhocas Eisenia foetida. Ambos os corantes foram fracamente tóxicos para Daphnia magna, porém o RB15 apresenta maior potencial tóxico em relação ao DB38. Os corantes DB38 e RB15 induziram malformações em larvas de zebrafish Danio rerio, caracterizadas por falha na inflação da bexiga natatória e alteração na cauda. Portanto, nossos resultados mostram a importância de se fazer não só a análise individual de corantes têxteis, mas também dos tecidos que os contêm. Além da necessidade de se desenvolver técnicas de tingimento mais seguras em relação à solidez da cor sob condições úmidas e as perdas de corante para o ambiente durante a etapa de fixação, indicando maior atenção ao estudo de efeitos sub-letais na avaliação do impacto desses compostos no ecossistema aquático.The fabrics dyeing began thousands of years ago and the commercial availability of dyes is increasingly. The Brazilian textile industry plays a role of high importance, highlighting among the main economic activities in the country. The dyeing process is one of the key factors in the commercial success of textile products, since consumers are demanding colors more resistant to heat, light exposure, perspiration and washing. According to the literature, conditions of intense perspiration contribute to the migration and subsequent penetration of textile dyes to human skin. Furthermore, 2 to 50% of the initial dye load is present in the dye bath effluent and these compounds are discharged in industrial effluents, contaminating the environment and endangering human health, since the wastewater treatment systems are ineffective in removing the color and mutagenicity of some dyes. In this context, this study aimed to evaluate the toxicogenetic effects of the Direct Black 38 (DB38) dye original and extracted by leaching with artificial sweat using Comet assay with fibroblasts and keratinocytes from human skin, Anexin V assay with fibroblasts and Salmonella mutagenicity test. Additionally, we investigated the ecotoxicity of textile dye Direct Black 38 and Reactive Blue 15 (RB15) using assays with seeds, dapnhias, worms and zebrafish performed in UTOX in Barcelona. The original and leached DB38 dye did not induce genotoxicity in fibroblasts and keratinocytes from human skin. The original DB38 was mutagenic for TA98 and TA100 of S. typhimurium with S9. However, the solution with the leached dye did not induce mutagenicity for these tested strains, since the highest migration rate of the dye to the solution of artificial sweat was ~ 1% in the following conditions: type of dyeing without rinsing, pH 8.0 and 8-hour incubation at 42°C. The original dye was cytotoxic for fibroblasts after 48 hours of exposure. However, this cytotoxicity was no longer observed after leaching in sweat. The original DB38 and RB15 dyes showed no toxicity for cucumber, lettuce and tomato seeds and for earthworms Eisenia foetida. Both dyes were weakly toxic for Daphnia magna, but the RB15 has a higher toxic potential compared to DB38. The dyes DB38 and RB15 induced malformations in larvae of zebrafish Danio rerio by failure of the swim bladder inflation and changes in the tail. Therefore, our results show the importance of making the individual analysis of textile dyes, but also of fabrics containing them. Furthermore, it is necessary to develop safer techniques of dyeing in relation to the color fastness under humid conditions and the loss of dyes into the environment during the fixation step, indicating more attention to the study of sub-lethal effects in the evaluation of the impact of these compounds in the aquatic ecosystem.
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Heyn, Tabea [Verfasser]. "Einfluss der glucosidierten Phospholipidanaloga Glc-PC und Glc-PAF auf die Proliferation, Zell-Matrix-Adhäsion und Migration in HaCaT-Zellen / Tabea Heyn." Berlin : Medizinische Fakultät Charité - Universitätsmedizin Berlin, 2011. http://d-nb.info/1025305418/34.
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Ebabe, Elle Etienne Raymond. "Le double aspect des nanoparticules manufacturées sur les métabolismes oxydatifs et inflammatoires : effets délétères et effets protecteurs." Thesis, Montpellier, 2016. http://www.theses.fr/2016MONTT008/document.
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On étudie les effets des nanoparticules (d'argent et de silice) manufacturées sur les métabolismes oxydatifs et inflammatoire. La première partie étudie la toxicité in vivo de l'ingestion de nanoparticules d'argent, pendant 11 semaines, sur un modèle animal - rat Sprague Dawley. Nous y avons mis en évidence l'action toxique des nanoparticules d'argent notamment une hausse de la production d'anion superoxyde par les NADPH oxydases hépatiques et cardiaques, des dyslipidémies, une cytolyse hépatique, une augmentation de cytokines pro-inflammatoires et une tendance à la baisse de l'activité d'enzymes antioxydantes. Ceci nous a conduit à aborder l'étude in vitro, sur des modèles cellulaires intestinaux (Caco-2) et cutanés (HaCaT). Au cours de cette étude, des nanoparticules de silice, fonctionnalisées ou non avec des antioxydants, ont été incubées pendant 24 H en présence des cellules. Nous montrons que la modification de la surface des nanoparticules réduit considérablement leur toxicité en limitant la production d'espèces radicalaires et la mortalité cellulaire. D’autre part, le couplage avec un antioxydant permet d’augmenter la stimulation de voie de signalisation du facteur Nrf2. Cette voie est impliquée dans la protection de l’organisme contre les troubles liés aux espèces radicalaires. En somme, ce travail met en avant les potentialités de la vectorisation d’antioxydants avec des nanoparticules à des fins thérapeutiquesThe purpose of this study is to explore the effects of nanoparticles (silver and silica) manufactured on oxidative and inflammatory metabolism. In the first part of this work, we explored the in vivo toxicity from ingestion of silver nanoparticles, for 11 weeks, in an animal model - Sprague Dawley rat. This enabled us to demonstrate the toxic properties of silver nanoparticles including superoxide anion production by hepatic and cardiac NADPH oxidases, dyslipidemia, hepatic cytolysis, an increase in proinflammatory cytokines and a downward trend the activity of antioxidant enzymes. This led us to address the in vitro study on intestinal cell models (Caco-2) and cutaneous (HaCaT). During this study, silica nanoparticles, functionalized or not with anti-oxidants, were incubated for 24 hours in the presence of the cells. We show that the modification of the surface of the nanoparticles significantly reduces their toxicity limiting the production of free radical species and cell death. Furthermore, the coupling with an anti-oxidant increases the stimulation of Nrf2 factor that involves the protection of the body against disorders associated with radical species. In summary, this work highlights the potential of vectorization of antioxidants with nanoparticles for therapeutic purposes
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Pothana, Kartheek. "Low dose UV-B induced keratinocyte exosomes protect Schwann cells against high glucose injury." Wright State University / OhioLINK, 2020. http://rave.ohiolink.edu/etdc/view?acc_num=wright1610380812991387.
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Charvat, Sandrine. "Influence de l'oncogène Ha-ras sur la migration cellulaire et la sécrétion de facteurs favorisant l'invasion tumorale dans les kératinocytes épidermiques humains, HaCat." Lyon 1, 1998. http://www.theses.fr/1998LYO1T248.
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Wu, Qiong. "The Role of Lipid Raft-Translocation of Prohibitin in Regulation of Akt and Raf-Protected Apoptosis of HaCaT Cells upon Ultraviolet B Irradiation." Ohio University / OhioLINK, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1382638433.
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KHALILPOUR, SABA. "VALIDATION OF PLANTS TRADITIONALLY USED FOR SKIN INFLAMMATION." Doctoral thesis, Università degli Studi di Milano, 2019. http://hdl.handle.net/2434/692911.
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Rhus coriaria L. (R. coriaria), Arctium lappa L. (A. lappa) and Echium amoenum Fisch. (E. amoenum) are medicinal herbs with extensive traditional uses, covering anti-inflammatory skin therapy. The present research aims to investigate the anti-inflammatory effects of different extracts of R. coriaria fruit, A. lappa roots and the E. amoenum flowers in human keratinocytes (HaCaT cells), evaluating extracts prepared using different techniques. Firstly, the herbal extracts were prepared using different methods including cold extraction, maceration, and decoction. The extraction solvents used were water, ethanol, ethanol-water (50:50), acetone, ethyl acetate, and dichloromethane. In the first phase of this study the extracts of E. amoenum, A. lappa and the polar (water (WRC), ethanol-water (EWRC), ethanol (ERC) and macerated ethanol (mERC)) extracts of R. coriaria were tested. The cytotoxicity and IL-8 release using, respectively, MTT and ELISA assays, were performed after 6 hours of TNF-α (10 ng/mL) treatment. Based on these results, the mERC and the EWRC were selected as the active extracts and assessed for NF-κB driven transcription, following identical treatment conditions. A non-toxic extract of A. lappa (EWAL) was also tested for its effect on NF-κB signaling. According to the preliminary data obtained, R. coriaria showed the most consistent and promising effects in inhibition of lL-8 and NF-κB signaling.Therefore, starting from this phase of the research the main focus of the study was on this herb and selectively mERC and the EWRC as the active extracts. The challenged cells by TNF-α, under treatment conditions with mERC and EWRC, were analysed for ICAM-1, VEGF, and MMP-9 releases, as well as NF-κB translocation, by ELISA assays. In addition, both active extracts were investigated for the inflammation induced by UVB (40 mJ/cm2) and were chemically profiled through HPLC-UV-DAD analysis. In the second phase of this research, the lipophilic extracts of R. coriaria (ARC, EARC and DCMRC) were evaluated using the cytotoxicity and IL-8 release assays in TNF-α or PMA-treated cells. Based on these results, active extracts were measured for their effects on NF-κB driven transcription. Among the evaluated herbs, only R. coriaria extracts achieved anti-IL-8 activity. Although all the polar extracts of this plant inhibited the TNF-α-induced IL-8 release, just mERC and EWRC suppressed NF-κB activation, ICAM-1, and MMP-9 secretion. EWRC showed higher inhibition on ICAM-1 and MMP-9 with IC50s of 1.76 ± 0.24 and 1.24 ± 0.33 µg/mL, respectively (mean ± s.d.). On the contrary, mERC significantly decreased VEGF levels whereas EWRC did not show any effect. The HPLC-UV profile of the extracts revealed higher amount of anthocyanins in EWRC in comparison with mERC. mERC, which showed lower amount of anthocyanins than EWRC, blocked both UVB-induced IL-8 release (with IC50 of 7.61 μg/mL) and translocation of NF-κB (at 10 μg/mL), measured by ELISA assay. The R. coriaria acetone (ARC) and ethyl acetate (EARC) extracts significantly inhibited the IL-8 release (at 25 μg/mL), as well as NF-κB driven transcription (at 50 μg/mL), when induced by TNF-α. The dichloromethane extract of this herb failed to show IL-8- inhibitory activity against TNF-α. ARC (with IC50s of 25.77 μg/mL ) and mERC (at 50 μg/mL) showed a significant effect in inhibiting the PMA induced IL-8 release. ARC modulated PMA-induced NF-κB signaling with IC50 of 27.82 μg/mL. Our results suggest the potential positive effect of R. coriaria fruit extracts, mostly mERC, as preventive agent in the treatment of keratinocyte inflammation through their inhibitory effect on the production of skin pro-inflammatory mediators. Our findings on the promising inhibitory effects of R. coriria on NF-κB signaling seem to confirm the traditional use of this herb as a remedy to treat skin inflammatory conditions. Its use as skin anti-inflammatory agent deserves further investigation.
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Yan, Jhih-Yin, and 顏志殷. "Comparative Proteomic Analysis of Differentially Expressed Proteins between HaCaT and TM deficiency HaCaT Cells." Thesis, 2009. http://ndltd.ncl.edu.tw/handle/46763342890599509014.
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碩士中國醫藥大學
醫學檢驗生物技術學系碩士班
97
Thrombomodulin (TM) is an endothelial cell surface glycoprotein that performs anticoagulant, cell-cell adhesion, and anti-inflammatory functions in various tissues. TM has been detected in platelets, megakaryocytes, monocytes, neutrophils, smooth muscle cells, it is also expressed in epithelial keratinocyte. But the function of thrombomodulin in epithelial tissue remains unclear. In this thesis, TM deficiency HaCaT (HaCaT-siTM) cell line was used to study the biological effects of TM on epithelial tissue. We found that TM regulated the cell morphology, proliferation and migration in HaCaT cells. Altered protein expressions between HaCaT keratinocytes and TM deficiency HaCaT (siTM) cells were identified by 2D-DIGE (2-D Fluorescence Difference Gel Electrophoresis) coupled with mass spectrometry. Difference gel electrophoresis (DIGE) is a form of gel electrophoresis where up to three different protein samples can be labeled with fluorescent dyes (Cy3, Cy5, Cy2) prior to two-dimensional electrophoresis. Total 1100 spots wae detected by typhoon 9200. Total of 25 proteins were significantly (P < 0.05,ratio＞1.5) changed in abundance between HaCaT and HaCaT-siTM cells. Seventeen proteins were successfully identified by MALDI-TOF/TOF. We showed that knock down TM expression could lead to the increased expression of RAD23 homolog B, elongation factor 1-delta, dUTPase, and proteasome subunit alpha type-1, whereas Annexin A3 and procathepsin D were being down-regulated the expression of. Annexin A3 and procathepsin D were further confirmed by Western blotting. In conclusion that TM could lead to keratinocytes transformation but the role of annexin A3 and procathepsin D in this process remains to be further studied.
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Hwang, Yu-Rong, and 黃于容. "Effect of anti-oxidant componds in HaCaT cells." Thesis, 2013. http://ndltd.ncl.edu.tw/handle/z25k5q.
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碩士國立中興大學
生命科學院碩士在職專班
101
There are two compounded whitening ingredients named ACTIWHITE LS 9808 and INDUXIN LS 9868 being used frequently in the cosmetic industry. Though their whitening function is widely evaluated, the relevant of antioxidant activity and DNA repair capacity have not been investigated. In this study , we found that Whitening Complex ACTIWHITE LS 9808 did not lead to apotheosis under a certain concentration, however it did not reduce ROS generation significantly. On the other hand, NDUXIN LS 9868 was not apoptotic under a certain concentration, however, it inhibited ROS generation with just a smidgen concentration,suggesting INDUXIN LS 9868 may prevent the oxidation and possess cell protection ability. Based on our experiment, Complex ACTIWHITE LS 9808 exhibited DNA self-repair ability,supporting by the up-regulation of phosphorylated p53. Therefore we propose that LS 9808 participate in DNA reapir in our system. In conclusion, INDUXIN LS 9868 reduces intracellular ROS generation and it may also involve in DNA repair process.
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Chen, Bowei, and 陳伯偉. "Mechanism of ultraviolet induced differenatiation of HaCaT keratinocytes." Thesis, 2013. http://ndltd.ncl.edu.tw/handle/34884618883169915204.
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碩士國防醫學院
生物化學研究所
101
HaCaT cells were used in this study to investigate the effect of ultraviolet (UV) on the differentiation of keratinocytes and the underlying mechanisms. It has been suggested that keratinocytes undergo terminal differentiation under high Ca2+ concentration and low temperature. During the isolation and selection of HaCaT cell line, the cells were grown in medium containing 0.03 mM Ca2+ at 38oC. However, after 50 passages used in this study, this cell lost its temperature and Ca2+ sensitivity. Indeed, under basal state, the growth and differentiation are indistinguishable in the presence or absence of extracellular Ca2+. Twenty four hr after UVB (10 mJ/cm2) irradiation, the growth of HaCaT cells decreased and the expression of K1 and K10 and the phosphorylation of ERK and Akt increased compared with control cells. Removal of extracellular Ca2+ blocked UVB induced increases of K1 and K10 expression and ERK and Akt phosphorylation, while it has no effect on those in control cells suggesting that UVB induced Ca2+ influx which in turn activated phosphorylation of ERK and Akt and the expression of K1 and K10. In addition, JNK phosphorylation is also induced by UVB which is insensitive to the removal of extracellular Ca2+. Co-treatment of EGF with UVB reversed the increases of K1 and K10 expression and ERK phosphorylation, while it has no effect on Akt and JNK phosphorylation suggesting the correlation between ERK phosphorylation and K1 and K10 expression. Co-treatment of TrpV4 channel blocker RN1734 with UVB had no effect on the action of UVB. Taken together, our results indicate that UVB promotes the differentiation of keratinocytes. This effect depends on Ca2+ influx which in turn activates phosphorylation of ERK and expression of K1 and K10. EGF reverses the phosphorylation of ERK and blocks the differentiation.
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Wu, Chia-Jung, and 吳佳蓉. "Chrysin Prevents UV-induced Damage on Human HaCaT Keratinocytes." Thesis, 2009. http://ndltd.ncl.edu.tw/handle/56798232678692184080.
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碩士輔仁大學
基礎醫學研究所碩士班
97
UV irradiation is one cause of skin aging and skin cancer. Numerous studies have demonstrated UV exposure result in phosphorylation of extracellular signal-regulated kinases (ERK), p38 kinase, and c-Jun NH2-terminal kinases (JNK) mitogen-activated protein kinases (MAPK) in epidermal keratinocytes. Moreover, UV exposure can induce cyclooxygenase-2 (COX-2) expression and lead to skin inflammation and further cause of skin cancer. What is more, UVB induces aquaporin-3 (AQP-3) down-regulation in human skin keratinocytes, influences water transporting, and causes the dehydration of skin. Chrysin, a flavone, has been proved its efficacy on antioxidant, anti-inflammatory and anti-tumor proliferation. In addition, chrysin has been demonstrated its photoprotective effect in UVA-irradiated human dermal fibroblast. However, little attention has been paid on the protective effect against UVB-induced injury in keratinocytes. Therefore, in this study we investigate underlying protective effect of chrysin on UV-induced damage on HaCaT keratinocytes. We found that chrysin significantly increased cell viability and attenuated the phosphorylation of MAPK on HaCaT keratinocytes after exposure to UVA or UVB. Furthermore, chrysin could decrease the COX-2 expression and reduce cell apoptosis after UV exposure. Chrysin also reversed UVB-induced down-regulation of AQP-3. These data demonstrate that chrysin can prevent UV-induced damage and may be beneficial in the prevention of UV-induced skin injury.
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Lee, Cheng-Hung, and 李政宏. "Study on Arecoline-Induced Apoptosis In Human Keratinocyte HaCaT Cells." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/87013013600903904350.
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碩士臺北醫學大學
醫學科學研究所
96
Betel quid chewing has become very popular in Taiwan in recent decades, especially among groups of mid to lower social-economic population. Betel nut is a powerful and energizing stimulant. Its ingredient can maintain body temperature and sustain laborers through long hours. It also provides a sense of community around the stands. While chewing betel quid has a temporal effect in soothing the spirit from fatigue, it also causes various diseases, such as cancer. Arecoline is the major component of betel nut which may cause cell mutation. Recent studies have indicated that arecoline can induce the production of inflammatory protein factors, cell apoptosis, and increase caspase protein concentration. In this study, we utilize different concentration of arecoline to evaluate cell viability and apoptosis of HaCaT keratinocytes via MTT assay, DNA fragmentation assay and Flow cytometry. Moreover, those HaCaT keratinocytes treated with betel quid components are further treated with or without antioxidant vitamin C and NAC so as to evaluate their effect in cytoprotection. The results show that treating HaCaT keratinocytes with indicated arecoline causes the production of reactive oxygen species. Futhermore, one can confirm the ability of inducing cellular apoptosis of arecoline regarding HaCaT keratinocytes. We also found it effective to inhibit cellular apoptosis by applying NAC to those HaCaT keratinocytes treated with arecoline. In this study, we demonstrated that arecoline, the ingredient of betel nut, can influence HaCaT keratinocytes proliferation ability. Cellular apoptotic inhibition was observed in HaCaT keratinocytes treating with NAC. Thus, we believe that some antioxidants can provide protective effects for betel quid chewing.
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Hipler, Christine. "Untersuchungen zum Einfluss von Cyclodextrinen auf das Proliferationsverhalten von HaCaT-Keratinozyten /." 2006. http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&doc_number=015452432&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA.
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簡佳雯. "Molecular basis of sodium arsenite tumorigenicity in human skin HaCaT cells." Thesis, 2003. http://ndltd.ncl.edu.tw/handle/20560242302942028516.
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Wen, Chong Pei, and 鐘佩紋. "Protective effect of bitter melon triterpenoids on UVB-irradiated HaCaT cells." Thesis, 2016. http://ndltd.ncl.edu.tw/handle/62078841439744248937.
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碩士國立臺灣師範大學
人類發展與家庭學系
104
Ultraviolet B (UVB) radiation plays a vital role in skin photodamage and photoaging. It causes serious inflammation and DNA damage of epidermis, which is the outermost viable layer of the skin and provides skin barrier function. In our previous study, methanolic leaf extract of wild bitter melon (WBM, Momordica charantia L. var abbreviata Seringe) shows anti-tyrosinase activity and significantly reduced UVB-induced reactive oxygen species (ROS) production in vitro. In this study, we investigated the protective effects of bitter melon triterpenoids on UVB-irradiated HaCaT cells. Two cucurbitane triterpenoids, Kuguacin R and 3, 7, 25-trihydroxycucurbita-5, 23-dien-19-al (TCD) were isolated from ethanolic leaf extract of WBM. HaCaT cells were pretreated for 24h with Kuguacin R or TCD prior to UVB irradiation (20-30 mJ/cm2), except for DNA fragmentation test (100 mJ/cm2). Our results showed that Kuguacin R and TCD inhibited UVB-induced cytotoxicity, and also diminished the productions of interleukin (IL)-1β, IL-6, and IL-8. Furthermore, both compounds significantly reduced phosphorylation of MAPKs, NF-κB activation, c-Jun, clyclooxygenase-2 protein levels, and the prostaglandin E2 production. In addition, both compounds inhibited caspase-3 activation and DNA fragmentation. Besides, Kuguacin R and TCD showed potential effects on percutaneous absorption in vitro. Our findings suggested that WBM triterpenoids, Kuguacin R and TCD may be beneficial for UVB-induced damage of keratinocytes and suggested its potential use in skin UV protection.
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Yang, Tzu-yen, and 楊子姸. "A Study of Proteomic Analysis of HaCat Cell after UVB Radiation." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/75505995795720720460.
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碩士美和技術學院
健康與生技產業研究所
98
Overdose of UV radiation could cause acute or chronic damages to the skin. Short-term UV radiation could cause sun burn and erythema; long-term UV radiation could cause cancers and accelerated aging. High-dose radiation causes the generation of reactive oxygen species which could trigger the activation of complex signaling pathways in skin cells, reduce the amount of glutathione, and reduce the activities of some protective enzymes such as superoxidase and catalase. The consequences could retard the synthesis of collagen in skin to cause skin photoaging. Conventionally the studies on damages caused by overdose UV radiation are carried out by examining the activities of oxidases or their changes at the molecular level. The changes at the total protein levels has not been examined. The purpose of this research is to establish a system which employ proteomics to observe the overall protein changes in skin cells caused by UV radiation. By using two- dimentional electrophoresis analysis, sixteen protein spots have been identified by LC/MS/MS. The levels of thirteen protein spots including keratin1, keratin 9, MTHSP75, Lamin B2, stomatin (EPB72)-like 2, and thioredoxin peroxidase increased. The level of one BIP protein decreased. RT-PCR and western blot analysis have been carried out on some proteins to confirm the expression difference in mRNA and protein level. The developed system could be used to evaluate potential antioxidants through the cell level reflecting their effectiveness closer to real conditions. The function of the antioxidants in the cells can be observed and the screening of the potential antioxidants used for cosmetics formulations could be done efficiently.
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Chen, Yu-Ting, and 陳郁婷. "Increased frequency of cancer-initiating cells in arsenic exposed HaCaT cells." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/37941869671915591483.
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碩士國立陽明大學
口腔生物研究所
98
Arsenic exposure is still a severe toxicological and pathological problem in humans. Epidemiological evidence has demonstrated that chronic arsenic exposure results in increased incidences of skin, lung, liver, and bladder cancers, but it mechanism of carcinogenesis is still unclear. A human keratinocyte chronically low-dose exposure to inorganic arsenic model has been established in this laboratory. In the tumorigenicity test A0, A2, and T4R2 cells showed different growth rate and tumor sizes in nude mice. These results may be explained by the presence of different frequency of cancer initiating cells (CIC) in A0, A2, and T4R2 cells. This is a question worth of further investigation. In this study, serum-free culture system was established to analyze the effect of epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) on cell growth of these cell lines. Under the condition of bFGF defined medium, spheroid formation was observed. The spheroid frequency increases in the cells previously exposed to low dose of arsenic i.e., A1, A2, and T4R2 cells. In low cell density serum-free culture, A0, A1, A2, and T4R2 cells form different types of colony. The cell morphology of typeⅠcolony is typical epithelial cell type, type Π colony a mixed type colony, and type Ш colony composed of small and compact cells. Through single cell colony tracking for 1 month, spheroids are formed from type Π and type Ш colonies, but not typeⅠ. To further characterize these different types of colonies, various types of A2 colonies were individually cloned and amplified in regular medium containing 10% fatal calf serum. In general, type Π and type Ш colonies showed higher spheroid formation ability than typeⅠin serum free medium with bFGF. The tumorigenicity of clones from different type colonies were evaluated by xenografted nude mice model. The preliminary data showed that type Π and type Ш colonies cells may form tumor, but required for further confirmation. Taken together, arsenic exposure to HaCaT cells resulted in increased frequency of spheroid forming cells, which may be considered as cancer initiating cells.
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Ho, Chia-Lin, and 何佳霖. "Ecto-Nucleoside Triphosphate Diphosphohydrolase Modulates ATP Signaling in Human HaCaT Keratinocytes." Thesis, 2013. http://ndltd.ncl.edu.tw/handle/44377345499732613808.
Full textAbstract:
博士國立陽明大學
微生物及免疫學研究所
101
Keratinocytes are the major building blocks of the human epidermis. In many physiological and pathophysiological conditions, keratinocytes release adenosine triphosphate (ATP) as an autocrine/paracrine mediator that regulates cell proliferation, differentiation, and migration. ATP receptors have been identified in various epidermal cell types; therefore, extracellular ATP homeostasis likely determines its long-term, trophic effects on skin health. The possibility that human keratinocytes express surface-located enzymes that modulate ATP concentration, as well as the corresponding receptor activation, in the pericellular microenvironment was investigated. We observed that the human keratinocyte cell line HaCaT released ATP and hydrolyzed extracellular ATP. Interestingly, ATP hydrolysis resulted in adenosine diphosphate (ADP) accumulation in the extracellular space. Pharmacological inhibition by ARL 67156 or gene silencing of the endogenous ecto-nucleoside triphosphate diphosphohydrolase (NTPDase) isoform 2 resulted in a 25% reduction in both ATP hydrolysis and ADP formation. Using intracellular calcium as a reporter, we found that although NTPDases hydrolyzed ATP and generated sustainable ADP levels, only ATP contributed to increased intracellular calcium via P2Y2 receptor activation. Furthermore, knocking down NTPDase2 potentiated the nanomolar ATP-induced intracellular calcium increase, suggesting that NTPDase2 globally attenuates nucleotide concentration in the pericellular microenvironment as well as locally shields receptors in the vicinity from being activated by extracellular ATP. Our findings reveal an important role of human keratinocyte NTPDase2 in modulating nucleotide signaling in the extracellular milieu of human epidermis.
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Chao, Chih-Hao, and 趙志豪. "Morphology Analysis of Suspension co-culture of Hs68 & HaCaT Cells." Thesis, 2013. http://ndltd.ncl.edu.tw/handle/43473368796350471225.
Full textAbstract:
碩士國立臺灣大學
醫學工程學研究所
101
We developed a new co-culture method mainly in co-culturing cells in suspension. Unlike the traditional method, cells cultured by this method will aggregate to a sphere and form a 3D structure, which compared to other systems like the hanging drop method and the hydrogel method, our system is simple, easy and good in long term culture. Our research used fibroblast and keratinocyte in co-culture, and we adopted Hs68 cell line from fibroblast and HaCaT cell line from keratinocyte. These cells are both from mature male foreskin, and the interaction existing between the two cells have been confirmed. We used the shared characteristics of Hs68 and HaCaT, that both will suspend in chitosan coated plate, to perform a suspension co-culture method. In this system, we can easily observe how each cells aggregate in time-lapse photography. This paper also discussed the aggregation process in suspension co-culture system, which was analyzed by a simple simulation. We’ve got a first step results in hands-on practice at present. Academically, we also adopted the theory about cadherin to explain our model design, thus we conducted some experiments in measuring cell-cell cohesion force to support our model.