Dissertations / Theses on the topic 'Hypoglycemic and hypolipidemic effects'

nÃo hÃ<br>The Protium heptaphyllum (almecegueira) exudes an amorphous resin consisting of four binary mixtures of triterpenoids, and the mixture of &#61537;,&#61538;-amyrin its major constituent. In folk medicine, the resin of Protium heptaphyllum is used as anti-inflammatory, gastroprotective, analgesic, expectorant and healing. The mixture of pentacyclic triterpenes &#61537;,&#61538;-amyrin (AB) (63:37) has gastroprotective, antipruritic, antiinflammatory and antioxidant properties. Experimental studies with pentacyclic triterpene compounds belonging to the groups ursan, oleanan and lupan showed inhibition of different enzyme systems intimately related to metabolism/absorption of carbohydrates and lipids. Thus the hypoglycemic and hypolipidemic activity of AB was evaluated in Swiss mice, the models of diabetes induced by streptozotocin (STZ) and alloxan (ALX) and hyperlipidemia induced by triton WR-1339 and hyperlipidemic diet (DH). AB (10, 30 and 100mg/kg) was able to reduce blood glucose in a model of acute treatment in STZ-induced diabetes, we observed two times (3 and 5 hours after administration of AB). In five days after treatment of diabetic animals by ALX, AB (30 and 100 mg/kg) also decreased hyperglycemia, an effect also observed for the use of glibenclamide (10 mg/Kg) in both models. But in normal animals, AB did not affect blood glucose, unlike the positive control glibenclamide (10 mg/kg). Although the model of diabetes for ALX, AB reduced total cholesterol (TC) serum at a dose of 100mg/kg, and triglycerides (TG) at doses of 30 and 100 mg/kg. In this same model, reduced the serum amylase AB (AB 30 and 100 mg/kg) and serum lipase (AB 100 mg/kg), a result also observed in normal animals, the same doses. In the Oral Glucose Tolerance Test (OGTT), AB (30 and 100 mg/kg) was shown to reduce blood glucose levels 60 min after administration of glucose, and AB (10, 30 and 100 mg/kg) in 90 min. In the model of hyperlipidemia by triton, AB (10, 30 and 100 mg/kg) significantly reduced the levels of TC and TG, in times of 24 and 48 h after administration of triton, an effect also observed for fenofibrate (200 mg/kg) used as positive control. The levels of HDL-c elevation experienced with the use of AB at all doses and times observed, as well as fenofibrate. In the model of hyperlipidemia by HD, AB (10, 30 and 100 mg/kg) controlled weight gain of animals receiving HD, as well as reduced TC, TG, LDL-c and VLDL-c, although the latter only in doses 30 and 100 mg/kg. These two doses were also effective in raising HDL-c. All doses of AB administered reduced the atherogenic index. All doses of AB also reduced the hepatic cholesterol in this model. All effects reported were similar to the positive control (fenofibrate 200 mg/kg). AB (10, 30 and 100 mg/kg) was able to significantly improve the antioxidant defenses of the liver, because it elevated the activity of hepatic SOD and CAT when compared to control high-fat diet, and raised the level of NP-SH in dose of 100 mg/kg, as well as reduced lipid peroxidation by decreasing the MDA, at all doses. Together, these results indicate that &#945;,&#946;-amyrin has hypolipidemic and hypoglycemic effect and deserves further evaluation in larger animal models that simulate chronic conditions of diabetes and dyslipidemia, in addition to research on their mechanism of action.<br>O Protium heptaphyllum (almecegueira) exsuda uma resina amorfa constituÃda de quatro misturas binÃrias de triterpenÃides, sendo a mistura de &#61537;,&#61538;-amirina o constituinte majoritÃrio. Na medicina popular, a resina de Protium heptaphyllum à utilizada como antiinflamatÃria, gastroprotetora, analgÃsica, expectorante e cicatrizante. A mistura de triterpenos pentacÃclicos &#61537;,&#61538;-amirina (AB) (63:37) possui propriedades gastroprotetora, antipruriginosa, antiinflamatÃria e antioxidante. Estudos experimentais com compostos triterpÃnicos pentacÃclicos que pertencem ao grupo ursano, oleanano e lupano mostraram a inibiÃÃo de diferentes sistemas enzimÃticos intimamente relacionados ao metabolismo/absorÃÃo de carboidratos e lipÃdios, deste modo, a atividade hipoglicemiante e hipolipemiante de AB foi avaliada, em camundongos Swiss, nos modelo de diabetes induzida por estreptozotocina (STZ) e por aloxano (ALX) e hiperlipidemia induzida por triton WR-1339 e dieta hiperlipidÃmica (DH). AB (10, 30 e 100mg/Kg) foi capaz de reduzir a glicemia num tratamento agudo no modelo de diabetes induzida por STZ, nos dois tempos observados (3 e 5h apÃs administraÃÃo de AB). Em tratamento apÃs cinco dias de animais diabÃticos por ALX, AB (30 e 100mg/Kg) tambÃm diminuiu a hiperglicemia, efeito este, tambÃm observado para o uso de glibenclamida (10mg/Kg) nos dois modelos. PorÃm em animais normais, AB nÃo alterou a glicose sanguÃnea, ao contrÃrio do controle positivo glibenclamida (10 mg/Kg). Ainda no modelo de diabetes por ALX, AB reduziu o colesterol total (CT) sÃrico na dose de 100mg/Kg, bem como os triglicerÃdeos (TG) nas doses de 30 e 100 mg/Kg. Neste mesmo modelo, AB reduziu a amilase sÃrica (AB 30 e 100 mg/Kg) e a lipase sÃrica (AB 100mg/Kg), resultado este observado tambÃm em animais normais, nas mesmas doses. No Teste Oral de TolerÃncia à Glicose (TOTG), AB (30 e 100 mg/Kg) mostrou reduzir a glicemia 60 min apÃs a administraÃÃo de glicose, bem como AB (10, 30 e 100 mg/Kg) em 90 min. No modelo de hiperlipidemia por triton WR-1339, AB (10, 30 e 100mg/Kg) reduziu de forma significativa os nÃveis de CT e TG, nos tempos de 24h e 48h apÃs a administraÃÃo do triton WR-1339, efeito observado tambÃm para fenofibrato (200mg/Kg) utilizado como controle positivo. Os nÃveis de HDL-c sofreram elevaÃÃo com o uso de AB em todas as doses e tempos observados, assim como o fenofibrato. No modelo de hiperlipidemia por DH, AB (10, 30 e 100mg/Kg) controlou o ganho de peso dos animais que receberam a DH, bem como reduziram CT, TG, LDL-c e VLDL-c, porÃm este Ãltimo somente nas doses de 30 e 100mg/Kg. Estas duas doses tambÃm foram eficazes para elevar o HDL-c. Todas as doses de AB administradas reduziram o Ãndice aterogÃnico. Todas as doses de AB tambÃm reduziram o colesterol hepÃtico neste modelo. Todos os efeitos relatados foram similares ao controle positivo (fenofibrato 200mg/Kg). AB (10, 30 e 100 mg/Kg) foi capaz de melhorar significativamente as defesas antioxidantes do tecido hepÃtico, pois elevou a atividade das enzimas catalase e superÃxido dismutase hepÃticas, quando comparado ao controle dieta hiperlipÃdica, bem como elevou o nÃvel dos grupos sulfidrÃlicos nÃo protÃicos na dose de 100mg/Kg, assim como reduziu a peroxidaÃÃo lipÃdica ao diminuir o malondialdeÃdo, em todas as doses. Em conjunto, esses resultados indicam que &#945;,&#946;-amirina possui efeito hipoglicemiante e hipolipemiante e que merece maior avaliaÃÃo futura em modelos animais que simulem situaÃÃes crÃnicas de diabetes e dislipidemias, alÃm de pesquisa de seu mecanismo de aÃÃo.

To examine the effects of Momordica charantia (MC) and cinnamon on glucose uptake and adiponectin secretion (AS) fat cells, 3T3-L1 adipocytes were treated with a water extract of cinnamon (CE) and three concentrations of MC water and ethanol extracts. The treatment combination of 0.2 mg/ml MC water extract and 0.5 nM insulin was associated with an increased glucose uptake into the cells (61%) and increased AS from the cells (75%). Without insulin, 0.2 mg/ml of CE increased glucose uptake (100%) and completely inhibited AS from the cells. Sub-optimal concentrations of insulin did not further enhance the CE activity and, in combination with 50 nM insulin, a dose-dependent decrease in glucose uptake was observed. The present results indicate that preferentially water-soluble component(s) in MC enhance the glucose uptake action of sub-optimal concentrations of insulin in 3T3-L1 adipocytes. This effect is accompanied by and may be a result of increased AS. CE increases glucose uptake in these adipocytes but inhibits AS.

Plant sterols (PS) are effective in reducing plasma lipid concentrations, however, few studies have examined their cholesterol lowering effects in type 2 diabetics. The objective was to assess whether PS consumption alters blood lipid profile in hypercholesterolemic subjects with and without type 2 diabetes. Fifteen control subjects (age = 55.1 +/- 8.5 yr and BMI = 26.9 +/- 3.0kg/m2) and fourteen diabetic subjects (age = 54.5 +/- 6.7 yr and BMI = 30.2 +/- 3.0kg/m2) participated in a double-blinded, randomized, crossover, placebo-controlled feeding trial. The Western diet included either 1.8g/d of PS or cornstarch placebo each provided over 21 d separated by a 28 d washout period. Subjects consumed only foods prepared in Mary Emily Clinical Nutrition Research Unit of McGill University. Total cholesterol (TC) decreased (p < 0.05) from baseline with PS for control and diabetic subjects by 9.7% and 13.6%, respectively. TC decreased (P < 0.05) from baseline with placebo for control and diabetic subjects by 10.9% and 11.6%, respectively. Non high density lipoprotein cholesterol (non-HDL-C) decreased (p < 0.05) from baseline with PS for diabetic subjects by 18.5%. Low density lipoprotein cholesterol (LDL-C) levels were reduced (p < 0.05) from baseline with PS for control and diabetic subjects by 14.9% and 29.8%, respectively. The reduction of LDL-C due to PS alone is greater with type 2 diabetics. There were no significant changes in HDL-C and TG across diets or treatments. It is thus concluded that PS consumption with diet enhances non-HDL-C and LDL-C reduction compared with diet alone in hypercholesterolemic individuals with and without type 2 diabetes. Demonstration for the first time that PS alone are more efficacious in lowering LDL-C and non-HDL-C in diabetic individuals compared to non-diabetics confirm the beneficial effects of PS to help prevent cardiovascular disease (CVD) for this high risk population.

Type 1 diabetes (T1D) is a metabolic disorder characterized by destruction of the pancreatic beta-islet cells leading to complete loss of insulin production. Gliclazide is used in Type 2 diabetes (T2D) to stimulate insulin production but it also has beneficial extrapancreatic effects which make it potentially useful in T1D. In fact, some T2D patients continue to use gliclazide even after their diabetes progresses to T1D since it provides better glycemic control than insulin alone. About 30% of a gliclazide dose undergoes enterohepatic recirculation which may contribute to the observed high interindividual variability in its pharmacokinetics. This may limit its efficacy in T1D especially since diabetes can disturb the gut microbiota and give rise to changes in bile composition and enterohepatic recirculation. Improving the absorption of gliclazide through the use of bile acids and probiotics may reduce this variability and improve the efficacy of gliclazide in T1D. The aim of this thesis was to investigate the interaction between the semisynthetic bile acid, 12-monoketocholic acid (MKC) and gliclazide in terms of pharmacokinetics and hypoglycemic effects in a rat model of T1D with and without probiotic pretreatment. A parallel ex vivo (Ussing chamber) study was carried out to investigate the mechanism of the interaction. Sensitive LC-MS and HPLC methods (Chapter 2) were developed to determine the concentrations of gliclazide and MKC in Ringer's solution and rat serum. Diabetes was induced in male Wistar rats by intravenous (i.v.) alloxan (30 mg/kg). Rats with blood glucose concentration > 18 mmol/l and serum insulin concentration < 0.04 [mu]g/l, 2-3 days after alloxan injection were considered diabetic. A total of 280 male Wistar rats (Chapter 3) were randomly allocated into 28 groups (n=10) of which 14 were made diabetic. Then 7 groups of healthy and 7 groups of diabetic rats were gavaged with probiotics (10⁸ CFU/mg, 75 mg/kg) every 12 hours for three days after which single doses of gliclazide (20 mg/kg), MKC (4 mg/kg) or the combination were administered either by tail vein injection (i.v.) or by gavage. The other 14 groups (7 healthy and 7 diabetic) were gavaged with saline every 12 hours for three days and then treated in the same way. Blood samples were collected from the tail vein for 10 hours after the dose and analyzed for blood glucose, serum gliclazide & serum MKC concentrations. Serum concentration-time curves for gliclazide and MKC were used to determine pharmacokinetic parameters. In the parallel ex vivo study (Chapter 4), 88 rats were randomly divided into 22 groups (n=4 rats per group, 8 chambers per rat), of which 11 groups were made diabetic. Of the 22 groups, 8 groups (4 healthy and 4 diabetic) were pretreated with probiotics as described above to study their influence on gliclazide and MKC flux, 8 groups (4 healthy and 4 diabetic) were used to investigate the interaction between gliclazide and MKC during transport, and 6 groups (3 healthy and 3 diabetic) were used to study the influence of selective inhibitors of the drug transporters Mrp2, Mrp3 and Mdr1 on gliclazide flux. 10 cm piece of the ileum was removed from each rat, the underlying muscle layer and connective tissue removed and the epithelial sheets mounted into Ussing chambers. Gliclazide, MKC or a combination were added to either the mucosal or serosal side and samples collected from both sides for 3 h to determine mucosal-to-serosal absorptive flux (Jss[MtoS]) and serosal-to-mucosal secretory flux (Jss[StoM]) of gliclazide and MKC as appropriate. In diabetic rats, gliclazide alone had no effect on blood glucose levels (Ch3, exp2) whereas MKC reduced it from 23 � 3 to 18 � 3 mmol/l (Ch3, exp3) and the combination of gliclazide and MKC reduced it even further from 24 � 4 to 16 � 3 mmol/l (Ch3, exp4). In diabetic rats, probiotic treatment reduced blood glucose by 2-fold (Ch3, exp1) and enhanced the hypoglycemic effect of the combination of gliclazide and MKC (blood glucose decreased from 24 � 3 to 10 � 2 mmol/l). The bioavailability of gliclazide was higher in healthy rats (53.2 � 6.2%) than in diabetic rats (39.9 � 6.0%) (Ch3, exp2). In healthy rats, MKC enhanced gliclazide bioavailability (82.7 � 8.2%) but in diabetic rats MKC had no effect on gliclazide bioavailability (Ch3, exp4). In healthy rats, probiotic pretreatment significantly reduced gliclazide and MKC bioavailabilities (p<0.01) while in diabetic rats, probiotic pretreatment significantly increased the low bioavailability of gliclazide to a level similar to that in healthy rats (Ch3, exp2 & 3). MKC showed clear evidence of enterohepatic recycling and probiotics delayed and reduced its systemic absorption (Ch3, exp3). In ileal tissues from healthy rats, Ussing chamber studies showed gliclazide is most likely a substrate of Mrp2 and Mrp3 (Ch4, exp5) and MKC significantly reduced gliclazide Jss[MtoS] probably through Mrp3 inhibition (Ch4, exp1). In ileal tissue from diabetic rats, MKC had no effect on gliclazide Jss[MtoS] and Jss[StoM] (Ch4, exp2) and none of the inhibitors had any effect of gliclazide flux (Ch4, exp6). This suggests that these transporters are dysfunctional in this model of T1D. Probiotics and MKC have hypoglycemic effects that appear to be enhanced by gliclazide and all appear to interact at the level of ileal drug transporters. The combination of probiotic treatment, gliclazide and MKC exerted the greatest hypoglycemic effect in T1D rats. Accordingly, the application of this combination may have potential in improving the treatment of T1D.

碩士<br>輔仁大學<br>營養科學系碩士班<br>103<br>Hedyotis diffusa (HD) is a well-known Chinese folk medicine. Previous research has shown that HD has many potentially beneficial health effects including antioxidant, anti-tumor and immunomodulatory activities. Recent studies found that oleanolic acid and ursolic acid, the main phenolic compounds in HD, can reduce blood glucose by inhibiting gluconeogenesis in diabetic rats. Therefore, the present study investigated the hypoglycemic effects of Hedyotis diffusa in streptozotocin-nicotinamide-induced diabetic rats. The HD extracts derived from various solvents (hot water, 50% ethanol and 95% ethanol) were used to inhibit the enzyme activities (including glucose-6-phosphatase, G-6-pase and phosphoenolpyruvate carboxykinase, PEPCK) in gluconeogenesis were assayed. The results showed that HD extracts of 95% ethanol possessed the best inhibitory capacities of key enzymes in gluconeogenesis and therefore was chosen for the subsequent animal experiment. Wistar rats were randomly divided into control group, diabetic group, low- or high-dosage of HD extracts (1%, 3% extracts in diet, respectively) and positive control group (metformin). In addition to control group, rats were induced to develop type 2 diabetes mellitus by intraperitoneal injection of streptozotocin- nicotinamide in combination with high-fat diet. After four weeks, the results showed that ethanol extracts from HD significantly decreased postprandial blood glucose, blood glucose levels after oral glucose tolerance test (OGTT) at the 60th minute, and inhibited enzyme activities in gluconeogenesis, including G-6-Pase, fructose 1,6-bisphosphatase and PEPCK. In addition, diabetic rats treated with HD extracts displayed significantly decreased triacylglycerol, total cholesterol and thiobarbituric acid reactive substances (TBARS) content in serum. In conclusion, the 95% ethanol extracts of HD significantly inhibit enzyme activities in gluconeogenesis in vitro and improve postprandial blood glucose by inhibiting gluconeogenesis and hypolipidemia in type 2 diabetic rats.

by Fok Ho Yin.<br>Thesis (M.Phil.)--Chinese University of Hong Kong, 2000.<br>Includes bibliographical references (leaves 114-120).<br>Abstracts in English and Chinese.<br>Acknowledgements --- p.ii<br>Abstract --- p.iii<br>槪論 --- p.v<br>List of Abbreviations --- p.vi<br>Chapter Chapter 1 --- Introduction --- p.1<br>Chapter Chapter 2 --- Toxicological studies on the effect of Ilex latifolia extract and its saponin-enriched Fraction --- p.19<br>Chapter Chapter 3 --- Hypoglycemic effect of Ilex latifolia extract and its saponin-enriched fraction --- p.51<br>Chapter Chapter 4 --- Hypolipidemic effect of Ilex latifolia extract and its saponin-enriched fraction --- p.78<br>Chapter Chapter 5 --- Conclusion --- p.109<br>References --- p.114

碩士<br>國立臺灣大學<br>食品科技研究所<br>85<br>The Hypolipidemic Effects of Pu-Erh Tea Lan-Chi, Lin. Graduate Institute of Food Science and Technology, National Taiwan University, Taipei, Taiwan, R.O.C. Abstract Pu-Erh tea is a fermented tea produced in the Yunan district in China. Recently, this tea has also gained popularity in Taiwan. In Pu-Erh tea, the bioactive ingredients can be originated from the tea leaves and their transformed products by the endogenous enzymes. Metabolites derived from the fermentation microorganisms such as Aspergillus, and biotransformation products originally derived from tea may also contribute to the potential biological activities of Pu-Erh tea. A previous study has indicated that Pu-Erh tea reduces plasma cholesterol and triacylglycerol in female Wistar rats. Meanwhile, the adrenalin-induced lipolysis is also stimulated, These findings suggest that uptake of Pu-Erh tea may ameliorate hypolipidemia and obiesty. In this study, the potential of an aqueous extract of Pu-Erh tea (PET) on cholesterol biosynthesis in vitro and lipid metabolism in vivo were elucidated With lovastatin as a positive control, the incorporation of 〔2-3H〕acetate and R-〔2-14C〕mevalonic acid into cholesterol by the cultured human hepotoma cells (HepG2) was chosen as the in vitro model to elucidate the potential of PET to inhibite cholesterol synthesis. In animal study, male hamsters and male Spsgue-Dawley rats fed with cholesterol (1.0 %, w/w) were chosen as animal models to elucidate the potential of PET to inhibit hepatic cholesterol synthesis, intestinal cholesterol absorption, and enetrohepatic circulation of bile acids. In vitro syudy showed that PET reduced cholesterol biosynthesis significantly (40 mg/mL PET showed 56 % inhibition) with the action mechanism of PET being similar to that of lovastatin (pre-mevalonate inhibition). In animal models, the results showed that PET inhibited cholesterol biosynthesis when compared to the control group. It is intresting because lovastatin is a product of Aspergillus and Aspergillus has often been found in the preparation of Pu-Erh tea. PET supplemention also reduced hepatic cholesterol content and the levels of serum cholesterol ,TG and FFA (p<0.05). The reduction of serum FFA and elevation of incorporation of isotope labelled acetate into TG in adipose tissue of hamsters suggested that PET stimulated insulin sensitivity. The fecal cholesterol secretion was increased in hamsters and rats fed with 1%, 1.5% and 2% PET. Cholesterol absorption in the intestine is decreased by reducing the solubility of cholesterol in mixed micelles. The finding that PET contained catechins further supported the observations in these two animal models. In comlusion, PET reduced liver cholesterol content and serum levels of cholesterol, TG, FFA. The fecal cholesterol contents were increased. Meanwhile, PET had the effect to inhibit cholesterol biosynthesis in vitro and in vivo.

碩士<br>中原大學<br>生物科技研究所<br>96<br>It has been recognized that many foods containing with some special components which can either reduce risk factors of a disease or enhance health benefits. However, people are very hard to ingest a same health food for years to obtain its benefit. The aim of the study was to investigate can a person eat various foods in a category with same health function, in stead of a same single health food? Eighty hamsters were divided into 8 groups, 8 each, and fed recommended dose of Chlorella-Cryptomonadales, chitosan, oat, fish oil, adlay, a cycle menu of these 5 food items, a mixture of 1/5 recommended doses of these 5 food items or control diet for 6 weeks. Serum lipids were determined after animals were sacrificed. The results showed that all the individual health food, cycle menu or mixture of these health food diets could reduce serum triacylglycerol (TG), total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C) and increase high density lipoprotein-cholesterol (HDL-C), comparing with control group. The results also revealed that there was no difference among the treatment groups, except fish oil group on HDL-C concentrations. The result demonstrated that a people can rotate taking any health food in the same functional category to obtain same health effect, and not necessary to ingest a same one every day.

碩士<br>高雄醫學大學<br>天然藥物研究所碩士班<br>93<br>Dioscorea tuber has been used in medical treatment and functional food for thousand years. In this study, hypoglycemic activity was detected in polysaccharide-enriched fractions (PLE) and hot water extracts (HWE) of five kinds of Dioscorea tubers cultivated in Taiwan (numbers and origins: TA01: Dioscorea alata L., TA02: D. alata L., TA03: D. alata L. var. purpurea (Roxb.) M. Pouch., TA04: D. persimilis Prain et Burk. and TA05: D. alata L.) using streptozotocin (STZ) induced diabetic mouse/rat models. Animals showing blood glucose levels 250~300 mg/dL 48 h after STZ treatment were selected for study. In STZ-diabetic mice model, treated with HWE of TA01, TA05, and PLE of TA01, TA02, TA05, respectively, for 14 days significantly (p<0.05) lowered the fasting blood glucose levels when compared with the STZ-diabetic mice group. According to the preliminary results, TA01 and TA05 showed the best hypoglycemic effect in those of five cultivated Dioscorea tubers. Furthermore, in STZ-diabetic rats model, glucose concentrations were increased in serum of diabetic animals in comparison to normal and were significantly improved after treatment with 300 mg/kg HWE of TA01、TA05 and 100 mg/kg PLE of TA01、TA05 p.o. for 7 days and 14 days (p＜0.001), respectively. In addition, all test drugs can reduce the productions of malondialdehyde (p＜0.01) and the ratio value of pancreas, kidney, liver weight to the body weight of rats (p＜0.05~p＜0.01). Otherwise, PLE of TA01 and TA05 showed the increasing of glycogen quantity in liver (p＜0.01~p＜0.001), and HWE of TA05 and PLE of TA01, TA05 increased insulin secretion in serum (p＜0.05~p＜0.01). According to the results, the hypoglycemic effect of PLE is better than that of HWE in Dioscorea tuber. Furthermore, TA01 and TA05 showed the best hypoglycemic effect in those of five Dioscorea tubers cultivated in Taiwan. The mechanisms of the hypoglycemic effect on Dioscorea tubers in this study were considered to the ability of increased insulin secretion and then enhanced the bioconversion of glucose to glycogen in the liver. Moreover, decreased levels of lipid peroxidation in Dioscorea tuber indicative of organs protection in diabetic rats. Based on the result of this study, TA01 and TA05 are suggested to be developed as functional food for treatment of diabetes.

Ho Hing Man.<br>Thesis (M.Phil.)--Chinese University of Hong Kong, 2001.<br>Includes bibliographical references (leaves 140-156).<br>Abstracts in English and Chinese.<br>Chapter Chapter 1 --- General introduction<br>Chapter 1.1 --- History of soybean --- p.1<br>Chapter 1.2 --- Health benefits of soybean --- p.2<br>Chapter 1.3 --- Introduction to flavonoids --- p.2<br>Chapter 1.4 --- Bioavailability of flavonoids from foods --- p.3<br>Chapter 1.5 --- Pharmacological effects of flavonoids and their glycosides --- p.4<br>Chapter 1.5.1 --- Anticarcinogenic activity --- p.4<br>Chapter 1.5.2 --- Antioxidative activity --- p.7<br>Chapter 1.5.3 --- Cardioprotective activity --- p.9<br>Chapter 1.5.4 --- Osteoprotective activity --- p.10<br>Chapter 1.5.5 --- Neuroprotective activity --- p.12<br>Chapter 1.5.6 --- Antiangiogenic activity --- p.12<br>Chapter 1.6 --- Soy leaves --- p.13<br>Chapter Chapter 2 --- Isolation and purification of kaempferol glycosides and genistin in soy leaves<br>Chapter 2.1 --- Introduction --- p.14<br>Chapter 2.2 --- Objectives --- p.15<br>Chapter 2.3 --- Materials and Methods --- p.16<br>Chapter 2.3.1 --- Extraction and isolation --- p.16<br>Chapter 2.3.1.1 --- Preparation of soy leaves butanol extract --- p.16<br>Chapter 2.3.1.2 --- Preparation of kaempferol glycosides from soy leaves butanol extract --- p.16<br>Chapter 2.3.2 --- High performance liquid chromatography (HPLC) analysis --- p.19<br>Chapter 2.3.2.1 --- Sample preparation for the HPLC analysis --- p.19<br>Chapter 2.3.2.2 --- HPLC analysis --- p.19<br>Chapter 2.3.2.3 --- Quantification of the flavonoids and their glycosides --- p.23<br>Chapter 2.3.2.4 --- Change in flavonoids and their glycosides in soy leaves --- p.23<br>Chapter 2.4 --- Results --- p.24<br>Chapter 2.4.1 --- Compound 1 --- p.24<br>Chapter 2.4.2 --- Compound 2 --- p.24<br>Chapter 2.4.3 --- Compound 3 --- p.25<br>Chapter 2.4.4 --- Compound 4 --- p.25<br>Chapter 2.4.5 --- Compound 5 --- p.25<br>Chapter 2.4.6 --- Compound 6 --- p.26<br>Chapter 2.4.7 --- Quantification of flavonoids in soybean and soy leaves --- p.32<br>Chapter 2.4.8 --- Age-dependent changes in flavonoids and their glycosides --- p.32<br>Chapter 2.5 --- Discussion --- p.35<br>Chapter 2.5.1 --- Compound 1 --- p.35<br>Chapter 2.5.2 --- Compound 2 --- p.35<br>Chapter 2.5.3 --- Compound 3 --- p.37<br>Chapter 2.5.4 --- Compound 4 --- p.38<br>Chapter 2.5.5 --- Compound 5 --- p.39<br>Chapter 2.5.6 --- Compound 6 --- p.40<br>Chapter 2.5.7 --- Age-dependent changes in flavonoids and their glycosides --- p.40<br>Chapter Chapter 3 --- Hypolipidemic effects of soy leaves in hamsters<br>Chapter 3.1 --- Introduction --- p.41<br>Chapter 3.1.1 --- Different lipoproteins and their functions --- p.41<br>Chapter 3.1.2 --- Risk factors of cardiovascular disease --- p.42<br>Chapter 3.1.3 --- Animal model --- p.43<br>Chapter 3.2 --- Objectives --- p.44<br>Chapter 3.3 --- Materials and Methods --- p.45<br>Chapter 3.3.1 --- Animals --- p.46<br>Chapter 3.3.2 --- Serum lipid and lipoprotein determinations --- p.46<br>Chapter 3.3.3 --- Determination of cholesterol in the liver and adipose tissue --- p.46<br>Chapter 3.3.4 --- Extraction of neutral and acidic sterols from fecal samples --- p.49<br>Chapter 3.3.4.1 --- Determination of neutral sterols --- p.49<br>Chapter 3.3.4.2 --- Determination of acidic sterols --- p.50<br>Chapter 3.3.4.3 --- GLC analysis of neutral and acidic sterols --- p.51<br>Chapter 3.3.5 --- Statistics --- p.51<br>Chapter 3.4 --- Results --- p.54<br>Chapter 3.4.1 --- Growth and food intake --- p.54<br>Chapter 3.4.2 --- "Effects of SLP and SLEE supplementation on serum triacylglycerol (TG), total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C)" --- p.54<br>Chapter 3.4.3 --- Effects ofSLP and SLEE supplementation on non-HDL-C and ratio of non-HDL-C to HDL-C --- p.55<br>Chapter 3.4.4 --- Effects of SLP amd SLEE supplementations on concentration of hepatic cholesterol --- p.58<br>Chapter 3.4.5 --- Effects of SLP and SLEE supplementations on perirenal adipose tissue cholesterol --- p.58<br>Chapter 3.4.6 --- Effects of SLP and SLEE supplementations on fecal neutral and acidic sterols --- p.61<br>Chapter 3.5 --- Discussion --- p.64<br>Chapter Chapter 4 --- Effects of soy leaves and its flavonoid glycosides on haemolysis and on LDL oxidation<br>Chapter 4.1 --- Introduction --- p.67<br>Chapter 4.1.1 --- Role of low density lipoprotein oxidation in the development of atherosclerosis --- p.68<br>Chapter 4.1.2 --- LDL oxidation --- p.70<br>Chapter 4.1.3 --- Thiobarbituric acid reactive substances (TBARS) as an index of LDL oxidation --- p.71<br>Chapter 4.1.4 --- Antioxidant and LDL oxidation --- p.74<br>Chapter 4.2 --- Objective --- p.75<br>Chapter 4.3 --- Materials and methods --- p.76<br>Chapter 4.3.1 --- Isolation of LDL from human serum --- p.76<br>Chapter 4.3.2 --- LDL oxidation --- p.77<br>Chapter 4.3.3 --- Determine the formation of thiobarbituric acid-reactive substances (TBARS) --- p.77<br>Chapter 4.3.4 --- Assay for erythrocyte haemolysis --- p.78<br>Chapter 4.3.5 --- Statistics --- p.79<br>Chapter 4.4 --- Results --- p.80<br>Chapter 4.4.1 --- Effects of three different soy leaves extracts and flavonoid glycosides on LDL oxidation --- p.80<br>Chapter 4.4.2 --- Effects of three soy leaves extracts and flavonoid glycosides on erythrocyte haemolysis --- p.80<br>Chapter 4.5 --- Discussion --- p.85<br>Chapter Chapter 5 --- Relaxing effects of soy leaves and its flavonoids<br>Chapter 5.1 --- Introduction --- p.89<br>Chapter 5.1.1 --- Smooth muscle contraction --- p.90<br>Chapter 5.1.1.1 --- Sliding filament mechanism --- p.91<br>Chapter 5.1.2 --- Intracellular mechanisms involved in the regulation of smooth muscle contraction --- p.92<br>Chapter 5.1.2.1 --- Voltage-gated Ca2+ channels --- p.92<br>Chapter 5.1.2.2 --- Protein kinase C (PKC) mediated smooth muscle contraction --- p.93<br>Chapter 5.1.2.3 --- Thromboxane A2 receptor-mediated calcium channel --- p.94<br>Chapter 5.2 --- Objectives --- p.96<br>Chapter 5.3 --- Materials and methods --- p.97<br>Chapter 5.3.1 --- Drugs preparation --- p.97<br>Chapter 5.3.2 --- Vessel preparation --- p.97<br>Chapter 5.3.3 --- Contraction experiments --- p.99<br>Chapter 5.3.3.1 --- Relaxant responses of soy leaves butanol extract on the contraction induced by different constrictors --- p.99<br>Chapter 5.3.3.2 --- Relaxant responses of soy leaves butanol extract on U46619 and PGF2a- induced contraction --- p.99<br>Chapter 5.3.3.3 --- "Relaxant responses of genistein, genistin and the kaempferol glycosides on U46619-induced contraction" --- p.100<br>Chapter 5.3.4 --- Statistics --- p.100<br>Chapter 5.4 --- Results --- p.102<br>Chapter 5.4.1 --- Effect of soy leaves butanol extract --- p.102<br>Chapter 5.4.2 --- Role of endothelium in extract-induced relaxation --- p.102<br>Chapter 5.4.3 --- Effect of the soy leaves butanol extract on contractile response to prostaglandins --- p.103<br>Chapter 5.4.4 --- Effects of kaempferol glycosides and kaempferol --- p.111<br>Chapter 5.4.5 --- Effects of genistein and genistin --- p.111<br>Chapter 5.5 --- Discussion --- p.118<br>Chapter Chapter 6 --- Effect of soy leaves on mammary tumor<br>Chapter 6.1 --- Introduction --- p.123<br>Chapter 6.1.1 --- Carcinogenesis --- p.123<br>Chapter 6.1.1.1 --- In itiation --- p.124<br>Chapter 6.1.1.2 --- Promotion --- p.124<br>Chapter 6.1.1.3 --- Progression --- p.125<br>Chapter 6.2 --- Objective --- p.126<br>Chapter 6.3 --- Materials and methods --- p.127<br>Chapter 6.3.1 --- Animal --- p.127<br>Chapter 6.3.2 --- Determination of estrus cycle --- p.128<br>Chapter 6.3.3 --- Statistics --- p.129<br>Chapter 6.4 --- Results --- p.131<br>Chapter 6.4.1 --- Incident rate of tumor induction --- p.131<br>Chapter 6.4.2 --- Number of tumor induced --- p.131<br>Chapter 6.5 --- Discussion --- p.136<br>Chapter Chapter 7 --- Conclusions --- p.136<br>References --- p.140

碩士<br>中州科技大學<br>保健食品系<br>102<br>Taiwan's five-leaf pine (Pinus morrisonicola Hay; PM) has been shown to have antioxidant, anti-inflammatory and hypolipidemic potential. Past studies show that the antioxidant and hypolipidemic effects of supercritical CO2 extract refraction from PM needles purified by thin layer and column chromatography methods (PME3-1) is better than unrefracting of PME3. Therefore, the aim of this study was further to investigate the hypolipidemic effects of PME3-1 and its possible mechanisms in vivo. We fed a high-fat and high-cholesterol diet (15% Fat, 0.2% Cholesterol)(HFC diet) for 30 days before feeding PME3-1 (low, medium and high doses of 0.2, 1.0 and 5.0 mg/kg bw) in 8-weeks-old hamsters for 60 days, and continuous feeding HFC diet during the period. The results indicated that supplementation with PME3-1 can significantly inhibit the liver and kidney swelling and restore the spleen weight of hamster induced by HFC diet. There were no significant difference in liver, kidney and spleen weight of hamster when compare with those of blank group(p>0.05). In the serum biochemical parameter analysis, cholesterol, triglycerides and LDL/HDL ratios in PME3-1-treated hamster were all significantly lower than those of the HFC diet group (p<0.05), and the inhibitory effects of PME3-1 on the serum lipidemia parameter were even better than that of Simvastatin (5 mg/kg b.w) (p<0.05). Histopathological evaluation of the hamster liver and kidney revealed that PME3-1 reduced the incidence of fatty change in liver and chronic progressive nephrosis in kidney induced by HFC diet. In addition, it found that high doses of PME3-1 group can significantly reduce total lipids (TL), total cholesterol (TC) and total triglycerides (TG) and inhibit HMG-CoA reductase activity of liver (p<0.05), and significantly promote fecal TC and TG (p<0.05) excretion in HFC diet hamsters (p<0.05). In the antioxidant ability assessment, the results indicated that PME3-1 can significantly reduce lipid peroxides- malondialdehyde formation of liver at the low dose of PME3-1, however, only high-dose of PME3-1 can significantly reduce glutathione peroxidase activity (p<0.05). Furthermore, the other enzymes, such as superoxide dismutase and glutathione S-transferase of liver cannot be influence by PME3-1 in HFC diet hamster. Based above results, PME3-1 revealed the potent hypolipidemic effects and the inhibitory ability on lipid peroxidation of liver in HFC diet hamsters. Thus, we speculated that hypolipidemic effects of PME3-1 may be due to the inhibition of TC and TG synthesis and reduction of lipid peroxidation in the liver. We suggested that PME3-1may be useful for prevention of cardiovascular disease.

碩士<br>輔仁大學<br>營養科學系碩士班<br>101<br>The global prevalence of diabetes mellitus (DM) is increasing rapidly in recent years, and disease ranked among the top ten leading causes of death in Taiwan. Glossogyne tenuifolia (GT), a native plant of Penghu, Taiwan, is usually used as an herbal tea. Recent researches have been shown that the active components in GT are potential inhibitors of α-glucosidase. The present study investigated that whether or not GT could improve the status of type 2 DM. The effects of GT extracts derived from various solvents (95% ethanol, 50% ethanol and hot water) on the inhibition of α-glucosidase activity in vitro were compared. The results showed that hot water extracts of GT possessed the best inhibitive capacities and therefore which was chosen for the following animal experiment. Male Wistar rats aged eight weeks were induced to be hyperglycemic by subcutaneous injection of streptozotocin (STZ, 65 mg/kg) –nicotinamide (NA, 230 mg/kg) (STZ–NA) and combination of high-fat diet (HFD). The animals were given GT extracts at low dose (50 mg/kg) or high dose (150 mg/kg), or the anti-diabetic drug (acarbose) in drinking water for 4 weeks. The results showed that hot water extracts from GT resulted in significantly decreased fasting blood glucose at the 1st and 2nd weeks, fasting insulin levels at the 2nd week, 1 hour postprandial blood glucose after starch loading test on the day 23 and blood glucose levels after oral glucose tolerance test (OGTT) at the 60th minute on the day 25. In addition, diabetic rats treated with GT extracts from hot water for 4 weeks displayed significantly decreased thiobarbituric acid reactive substances (TBARS) in the serum, liver and kidney, serum total cholesterol, fasting insulin levels and homeostasis model assessment for insulin resistance (HOMA-IR). In conclusion, the hot water extracts of GT exhibited α-glucosidase activity in vitro, and might improve the progression of diabetes and decreased oxidative stress in STZ–NA-induced diabetic rats.

碩士<br>國立屏東科技大學<br>生物科技研究所<br>98<br>The occurrence of type 2 diabetes is closely associated with the development of insulin resistance. Previously, two triterpenes, CH63 and CH93, were isolated from the extract of the stem of Momordica charantia. They were proved to promote glucose uptake, activate the insulin signaling pathway and AMP-activated protein kinase (AMPK) in insulin-resistant cells. Therefore, CH63 and CH93 were considered to be hypoglycemic molecules. The purpose of this study is to further explore the molecular mechanisms underlying the hypoglycemic activitise of CH63 and CH93. In this study, it was shown that CH63 and CH93 can promote the translocation of glucose transporter 4 (GLUT4) to the cell membrane in FL83B cells, which is likely an important mechanism for their hypoglycemic effects. Meanwhile, the data suggested that the activation of AMPK plays a key role for the hypoglycemic effects of CH63 and CH93. Furthermore, the data indicated that the activation of AMPK by CH63 and CH93 was likely mediated by LKB1. Based on the data presented in this study, CH63 and CH93 were proposed to work as both insulin sensitizers and insulin substitutes.

碩士<br>大仁科技大學<br>藥學系碩士班<br>107<br>Diabetes is a metabolic disease. Diabetes is characterized by the blood sugar of the patient is over the standard value for a long time. At least 150 million people worldwide suffer from diabetes. This incident shows that diabetes has seriously threatened the health of the people. Currently, about 90% of patients with diabetes are classified as type 2 diabetes. In recent years, studies have shown that type 2 diabetes causes inflammatory factors such as TNF-α, IL-6 and IL-1β...etc, which produce an inflammatory response that in turn causes insulin resistance. In the normal insulin signaling pathway, insulin binds to the insulin receptor and phosphorylates IRS1/2/3, PI3K and Akt. Eventually, GLUT4 is displaced onto the cell membrane and glucose is transported into the cell for utilization. PTP1B is a negative regulator of the insulin signaling pathway. PTP1B exacerbates hyperglycemia in the case of insulin resistance. Cyclocarya paliurus is commonly used as a clinical hypoglycemic agent in traditional Chinese medicine. It is rich in a variety of chemical ingredients, which not only can lower the three highs, oxidative stress and inflammatory response. This study focused on the effects of Cyclocarya paliurus leaf-extract (CPE) on the regulation of blood sugar and inflammation. First of all discovered by the test of RINm5F cell proliferation, after addition CPE 50.0 μg/mLand 24 hours, the optimal proliferation effect (71.0%) was achieved. In the mouse of blood sugar regulation test, after continuous administration of CPE for 28 days, The CPE medium dose group (200.0 mg/mL) significantly increased Total-protein (14.99%), decreased Glucose AC (65.48%), Triglyceride (28.33%) and Total-cholesterol (31.32%); increased the performance of IRS2 (108.43%), Akt1/2/3 (93.18%) and GLUT4 (140.22%), and significant statistical differences (p<0.05) compared with the STZ group. In addition, compared with the TPA group, the reduction of ear edema was 26.9 % and 27.2 % respectively, after administration of high dose CPE (25.0 mg/mL) for 12 and 24 hours in the anti-inflammatory experiment of mice. In addition, compared with the TPA group, the reduction of ear edema was 26.9 % and 27.2 % respectively, after administration of high dose CPE ( 25.0 mg/mL ) for 12 and 24 hours in the anti-inflammatory experiment of mice, and the inhibitory effect is better than the positive control Medicine (Indomethacin). In the PTP1B inhibition test, the PTP1B content decreased with the higher dose. At the high dose of CPE (5.0 mg/mL), it has the lowest concentration (0.001 ± 0.08 nmol). It shows that the higher CPE concentration has the better inhibitory effect. Based on the above experimental results, CPE can improve the hyperglycemia of diabetic patients by regulating biochemical indicators and enhancing the transmission of insulin signals, and reduce the insulin resistance of patients by anti-inflammatory reaction. In addition, it can enhance insulin signal transmission by inhibiting the action of PTP1B. CPE can regulate blood sugar in many ways, which is beneficial to the recovery of diabetes.

碩士<br>大葉大學<br>生物產業科技學系<br>104<br>Cajanus cajan (L.) Millsp. root cooked with the ribs can cure diabetes is recognized an aboriginal traditional therapies. Previous studies known after a series of separation and purification from ethanol extract of Cajanus cajan root was obtained Betulinic acid, biochanin A, 5,2'-dihydroxy-7,4'-dimethoxyisoflavanone (cajanol), genistein, 2'-hydroxygenistein and other differences flavonoids. Many studies have confirmed the above components such as: genistein has excellent antioxidant capacity and the ability to inhibit the decomposition of carbohydrates. We investigated whether ethanol extracts of Cajanus cajan roots (EECR) could protect against methylglyoxal (MGO; 500 mg/kg bw)-induced insulin resistance (IR) in male Wistar rats between days 1 to 83. Rats treated with MGO were used to examine the hypoglycemic effects of EECR on IR. The rats were divided into six groups and orally supplemented with MGO except for group 1 (normal controls). Group 3 was orally supplemented with Metformin (MET;10 mg/kg bw), group 4 with EECR-L (10 mg/kg bw), group 5 with HIP EECR-M (50 mg/kg bw), and group 6 with EECR-H (100 mg/kg bw). MET and EECR were provided daily between days 31 to 83 in rats. Oral glucose tolerance (OGTT) and insulin tolerance (ITT）tests were carried out on days 77 and 78, respectively. The results indicated that body weights, water and food intake for each group revealed no significant difference. In insulin tolerance tests (ITT), serum glucose levels of MGO-treated group was slightly change (approximately 100 mg/ml) (no insulin sensitivity) after intraperitoneal injection of insulin, while serum glucose level was significantly decreased (P <0.05) in EECR-treated rats, the effects were comparable MET groups (MET is a known hypoglycemic drugs). Blood sugar and glycosylated hemoglobin (HbA1c) as well as serum insulin, glycation end products (AGEs) of MGO-treated groups were higher than that of blank groups. EECR significantly decreased those hyperglycemic index induced by MGO. In addition, EECR also dose dependently decreased serum biochemical index (such as, GOT, GPT, CHOL, LDL, TG), TBARS content and increased SOD activity. MGO can induce IR (increase the insulin and blood sugar level) in rats, whereas EECR could inhibit AGEs formation by reducing oxidative stress, so as to improve IR and lowering blood sugar. Thus, EECR has the potential to become a useful hypoglycemic agent for diabetic mellitus.

碩士<br>國立中山大學<br>生物科學系研究所<br>93<br>Recent studies indicated that Atherosclerosis was thought to the resulted from high blood lipid and oxidative stress. Dioscorea has been recognized as an edible medicinal herb as well as healthy plant. Related studies showed that Taiwannative yam (Dioscorea) exhibited hypoglycemic, antioxidative and hypolipidemic functions. However, researches regarding the best species of native Dioscorea revealing the efficacy with both antioxidative and hypolipidemic functions are limited. Previously our laboratory, found that three in five native species of Dioscorea provided by the Agricultural Research Institute had antioxidative effects. The goal of this study was to investigate the effects of this three Dioscorea species (TA01、TA03、TA05) on lipid metabolism and antioxidative ability of hamsters. Hyperlipidemia hamsters induced by feeding high fat and high cholesterol diet for 3 weeks, were randomly divided into five groups. Group 1 was the control group fed with a high fat and high cholesterol diet (HF, 0.2% cholesterol and 12% fat) ; group 2 was the positive control group fed with a HF diet supplemented with 0.08% atorvastatin (HF+ATS) ; group 3 was TA01 group fed with a HF diet supplemented with 10% TA01 yam powder (HF+TA01) ; group 4 was TA03 group fed with a HF diet supplemented with 10% TA03 yam powder (HF+TA03) ; group 5 was TA05 group fed with a HF diet supplemented with 10% TA05 yam powder (HF+TA05). The experiment was conducted for 12 weeks. Blood was collcted to determine serum total cholesterol (TC) and triglycerol (TG) at week 4 and 8. The hamsters were scarified and the blood, liver and epididymal fat pads were collected at week 12. The weight of body, liver, epididymal fat pads ; Serum TC, TG, high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and oxidation of LDL (thiobarbituric acid reactive substances, TBARS) were determined ; TC, TG, TBARS, superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx) and glutathione reductase (GR) of liver were also analyzed. Results indicated that TA01 group had significantly decreased serum TG (P < 0.05) at 4 and 8 weeks. Serum TC and TG of TA03 group were significantly lower than those of the control group at 8 weeks. No significant difference for TC and TG was found between TA05 and the control groups. After 12 weeks of Dioscorea feeding, liver weight and liver to body weight ratio of TA01 and TA03 groups compared to that of control group were significantly reduced. Epididymal fat pads weight and that the body weight ratio tended to be less compared to that of the control group. Serum TG concentration of TA01 was significantly decreased to 89% of the control group. Serum TC, TG and LDL-C concentrations of TA03 group were significantly decreased to 82, 89 and 83% of the control group, respectively. Serum HDL-C of TA03 group was unaffected and LDL TBARS tended to be decreased (1.67 ± 0.79 vs. 1.06 ± 0.57) compared to that of the control group. Liver TC concentrations of TA01 and TA03 groups both were significantly decreased to 88% of the control group. Liver TG and TBARS of TA01 and TA03 groups were unchanged. Catalase activities of TA01 and TA03 groups and GR activity of TA01 group were significantly higher than those of the control group. No significant differences were found in GPx and SOD activities among all groups. In conclusion, TA01 and TA03 species of Taiwannative yam (Dioscorea) tubers exhibit beneficial effects on lipid profile and antioxidative status for hyperlipidemia induced by a high fat and high cholesterol diet. Among there species, TA03 reveals the best potential with both hypolipidemic and antioxidant effects, and was suggested to be used in the development of functional food for the modification of lipid profile and oxidative status in cardiovascular disease prevention .

碩士<br>臺灣大學<br>食品科技研究所<br>98<br>Wild bitter melon is traditionally known for its medicinal properties such as antioxidant, anticancer, hypoglycemic, and cholesterol lowering effects. We explored the effects of heating treatments and media milling on antioxidative and hypoglycemic activities of bitter melon. Antioxidative activity was evaluated by cellular antioxidant activity assay along with other methods. Hypoglycemic activity was evaluated by glucose uptake activity. In antioxidant activity assays, the results showed that the antioxidant activities decreased as heating temperatures being increased. Prolonging heating time at 65℃ resulted in low antioxidant activity. There was no significant difference in the antioxidant activity among all the samples being heated at 95℃ with different heating time. Comparing to media milled samples, both blended and milled seeds had the higher antioxidant activities. The result of polyphenol content indicated that polyphenol content decreased from 20.00 to 13.35 mg CAE/g with increasing heating temperature. The polyphenol content decreased from 20.00 to 12.36 mg GAE/g at 65℃ with increasing heating time, however there was no significant difference in the polyphenol content of samples heated at 95℃ with different heating time. Both blended seed (23.65 mg GAE/g) and media milled seed (21.19 mg GAE/g) had higher polyphenol contents than blended and milled fruits (20.00 and 14.68 mg GAE/g) . Tumor necrosis factor-αwas used to induce insulin resistance of mouse liver FL83B cells in vitro. Insulin-resistance cells was used to observe glucose uptake activity, blended fruit, milled fruit and seed significantly increased glucose uptake in TNF-α-induced insulin resistance FL83B cells, especially milled-seed. The positive effect might be attributed to higher bioactive compounds in milled seed. The content of charantin in samples was following order: milled seed(84.49 ug/g)＞ blended (68.79 ug/g)＞blended fruit(56.04 ug/g) ＞milled fruit (48.92 ug/g). The content of charantin decreased with the heating temperature increased. Based on our findings, the bioactive compounds of bitter melon fruit were influenced by heating process. The milled seeds showed higher antioxidant activity and enhanced glucose uptake activity.

碩士<br>國立中興大學<br>食品暨應用生物科技學系所<br>106<br>Prunus mume Sieb. et Zucc. is the deciduous tree of the Rosaceae family. It has antitussive, expectorant, antiemetic, and antipyretic effects in Chinese traditional medicine. It provides many physiological activities, including anti-obesity, anti-oxidation, anti-osteoporosis, hypoglycemic, anti-fatigue, and anti-cancer. Mei-Gin is the concentrated extract of plum fruit. However, the functional components and physiological effects of the plum fruit are destroied during the long time heating process. Therefore, this research used Mei-Gin formulas which are based on Mei-Gin and added different proportions of plum fruit, black garlic, and Mesona procumbens Hemsl. extracts to prepare seven kinds of Mei-Gin formulas. High-fat/cholesterol diet was used to induce hamsters with hyperlipidemia and the potential on hypolipidemia of these formulas was evaluated. The content is divided into two parts: (i) An animal pretest to screen the most potential sample from different Mei-Gin formulas. (ii) An animal test to evaluate the hypolipidemic effect on different doses of Mei-Gin formula 4. The results of animal pretest showed that Mei-Gin formula 4 could decrease the concentration of serum triglyceride, total cholesterol, and LDL-C. Furthermore, it also reduced the accumulation of hepatic total lipid and promoted the excretion of fecal cholesterol. From these results, Mei-Gin formula 4 was the most effective formula on improving hyperlipidemia. The results of animal test showed that treatment by Mei-Gin formula 4 (100 mg/kg b.w.) could significantly reduce the concentration of serum triglyceride, total cholesterol, and malondialdehyde. Stimultaneously, it also decreased the amount of hepatic total lipid and triglyceride. Besides, it was able to increase the release of fecal total lipid. We explored the hepatic tissue genes of hyperlipidemic hamsters and found that Mei-Gin formula 4 not only promoted the expression of LXR α, ABCG5, ABCA1, and CYP51, but also inhibited the expression of ACC, FAS, DGAT-2, HMG-CoA reductase, SREBP-1a, and SREBP-1c. In addition, it increased the richness and diversity of the gut microbiota in hyperlipidemia hamsters. Also, it increased the number of Lactobacillus acidophilus and Bifidobacterium animalis, and reduced Helicobacter pylori abundance, which could improve cholesterol metabolism and inhibited its synthesis to reach hypolipidemic effect. Based on these results, Mei-Gin formula 4 had the function to improve dyslipidemia in high-fat/cholesterol-diet-induced hamsters. It would be used to develop into health food to regulate serum lipid profiles in the future.

碩士<br>高雄醫學大學<br>醫學研究所<br>89<br>Toona sinensis (TS) is a broadleaf tree, which grows all around the world, which is composed of only four or five ‘good’ species. TS are the only Toona species in which the leaflet margins can be serrate to serrulate. The trees provide very high quality timber and are widely used medicinally, the bark being used as an astringent and depurative, the root as a refreshment and a diuretic, the tender leaves as a carminative and a corrective, and the fruits as an astringent and for treatment of eye infections. The leaves and young shoots have been used as a vegetable in China for thousands of years. The leaves and stems of this plant have been used for the treatment of enteritis, dysentery and itch in oriental medicine. Recently, Aqueous leaf extracts of TS was used as a folk medicine for lowering blood pressure and blood sugar in the U.S.A. and Taiwan. It is said that TS leaf aqueous extracts can reduce the fasting blood glucose (FBG) level of ill-controlled NIDDM patients who were treated with conventional medicines, but don’t affect the FBG level of a healthy person. However, the mechanism of hypoglycemic effect of TS on diabetes is still obscure. Alloxan causes severe damage in pancreatic b cells damage in rats. STZ cytotoxyicity is highly specific to the b cell, it also causes functional abnormalities in the islet a and d cells. Epinephrine increases the hepatic output of glucose and glycogenolysis. Testing with these three different diabetic or hyperglycemic animal models may provide us with the different possible mechanisms involved in hypoglycemic effect of TS. We found that aqueous leaf extracts of TS can lower the FBG level in alloxan-induced diabetic rats, while in STZ-induced and Epinephrine- induced diabetic rats the lowering effect was not shown. Glucose transporter 4 (GLUT4), an insulin-dependent glucose transporter, is wide spread in adipose tissue and muscle. It translocates to the membrane when activated by insulin. Thus, the present study was designed to compare the effects of TS and Glibenclamide (GC) and find out if TS affects the secretion of insulin. Moreover, the possible role of membrane-associated GLUT4 in participating the hypoglycemic effect of TS on diabetes was investigated. Animal model of Alloxan-induced diabetes was used. The incidence of Alloxan-induced diabetes was more than 75%. We found that TS lowered the blood sugar level only in diabetic rats in a time-dependent manner, but GC lowered fasting blood glucose levels in both diabetic and normal rats. In addition, TS improved the glucose intolerance of diabetic rats the same as GC. In our preliminary results, the serum level of insulin was increased by TS treatment. The protein and mRNA expression of GLUT4 glucose transporter in adipose tissues, but not in skeletal muscle and heart, were significantly decreased in diabetic rats. TS or GC treatment caused a significant increase of GLUT4 protein and mRNA content in white and brown adipose tissues. The liver plays an especially important role in regulating glucose by taking up incoming glucose from the small intestine and by releasing glucose into the circulation. Both uptake and release of glucose require transport of glucose across the plasma membrane. In general, specific carriers termed glucose transporters mediate glucose transport across the plasma membrane, and the presence of a facilitative glucose transporter, GLUT2, was suggested in the liver. In our preliminary results, the expression of GLUT2 glucose transporter was unchanged. These results show that TS might enhance the secretion of insulin by an unknown mechanism and GLUT4 protein and mRNA of adipose tissues might participate in the hypoglycemic effect of TS on diabetic rats. II英文摘要………………………………………………… 4-6 III緒論 香椿 …………………………………………………. 7-8 糖尿病………………………………………………… 9-12 葡萄糖轉運裝置(GLUT)……………………………… 13-14 GLUT4及GLUT2與糖尿病的關係……… 15-16 研究動機 .. …………………………………………. 17 IV實驗方法 實驗動物………………………………………………. 18 誘發糖尿病及高血糖之動物模式…………………….. 19-21 香椿葉萃取法…………………………………………. 22 最大耐受量測定(安全限度試驗)……………………… 23 血液中葡萄糖的濃度…………………………………. . 24 血中胰島素測定……………………………………….. 24-25 口服葡萄糖耐量試驗…………………………………… 26 細胞膜表面葡萄糖轉運裝置表現的測定…………….. 26-32 資料分析…………………………………………………… 33 V實驗結果 壹. 基本表現 1. 誘發糖尿病及高血糖動物模式 (A) 配製藥物注意事項………………………………. 35-36 (B) 治療前後之空腹血糖變化………………………. 36-37 (C) 動物評估………………………………………… 37-38 2. 外在因素對糖尿病鼠血糖值的影響……………… 38-39 3. 椿葉粗萃取粉劑的最大耐受量測定(安全限度試驗)… 39-40 4. 椿葉粗萃取粉劑與Glibenclamide對正常鼠及Alloxan誘 發的糖尿病鼠各項實驗結果 (A) 體重、飲水量、飲食量、排便量的變化…………. 40 (B) 空腹血糖 (1) 治療前血糖之穩定度……………………………. 41 (2) 治療後血糖的變化………………………………. 41-45 (C) 血中胰島素濃度……………………………………. 45-46 (D) 口服葡萄糖耐受性…………………………………… 46-47 貳.機轉探討 細胞膜表面葡萄糖轉運裝置的表現 (A) GLUT4蛋白質的表現………………………………… 48-49 (B) GLUT2蛋白質的表現………………………………… 49 (C) GLUT4 mRNA的表現………………………………… 50-51 參.附圖……………………………………………………… 52-79 VI討論……………………………………………………….. 80-89 VII參考文獻………………………………………………… 90-98

碩士<br>中國醫藥學院<br>醫學研究所<br>91<br>四 英文摘要 Cardiovascular disease is a common phenomenon when the diet contains high cholesterol. In this study the antioxidative and hypolipidemic effects of Chinese herb and anti-oxidant agent were investigated in hypercholesterolemic rabbit . The normal group was fed regular chow and the cholesterol group was fed a chow containing 0.5 % cholesterol. The Chinese herb group were fed 1 % herbal essence and were fed 0.1% anti-oxidant agent in antioxidant group . The total cholesterol, LDL-cholesterol, triacylglycerol and lucigenic-CL were decreased in the Chinese herb and anti-oxidant agent when compared with the cholesterol group. The intimal surface of the thoratic aorta was covered with atherosclerotic lesions in the control group, but reduced in the Chinese herb and antioxidant group. Therefore , Chinese herb may be useful to prevent cardiovascular disease in which atherosclerosis plays a major role.

碩士<br>國立宜蘭大學<br>食品科學系碩士班<br>101<br>Ganoderma lucidum is one of the most important Chinese medicinal and edible mushrooms. The most important active components of G. lucidum are polysaccharides and triterpenoids. They have been proven to possess many pharmacological functions such as antioxidant, anti-hepatitis, hypoglycemic activity, anti-inflammation and hypercholesterolemia. Trivalent chromium can improve blood sugar control and glucose tolerance in diabetes. Therefore, chromium with different concentrations (0, 100 and 200 ppm ) were added to brown rice medium with 50% moisture content for Ganoderma lucidum in a 14-day solid-state fermentation at 30℃. Then the Ganoderma lucidum fermented rice products were extracted by hot water or 85% ethanol, and their antioxidant activities were analyzed. Ganoderma lucidum fermented rice supplemented with 200 ppm chromium had higher contents of polysaccharides, total phenols, ferrous ions and higher reducing power. Different ethanol concentrations (0, 25, 50, 75 and 95%) were used for extracting chromium enriched Ganoderma lucidum fermented rice products by 300 W microwave for 10 min, and 25% ethanol obtained more active compounds, such as 19.75% crude polysaccharides, 0.34% crude triterpenoids, and 0.0026% total chromium. The mice were divided to a normal group, four STZ-induced mice groups including control group, 50 ppm GL extracted by 25% ethanol, 200 ppm GL polysaccharides with 3 ppm Cr+3, and 3 ppm Cr+3 groups. During 28-days feeding, water intake, feed intake and plasma glucose of mice were recorded. The feed and water intake significantly decreased in STZ-induced mice groups with 50 ppm GL ethanol extract after 14-day and 21-day, respectively; and the hypoglycemic effects were appeared 28-day after feeding. Rice pastas were made from 2% chromium enriched Ganoderma lucidum fermented rice products, different ratio of rice flour (50, 60, 70%), and corn starch. The powders were premixed and humidified before entering the uniaxial extruder. The rice pastas only required 7 min cooking, and the cooking loss and cooking yield were about 6-7.5% and 248-270%, respectively. The seven score hedonic sensory evaluation at appearance, color, flavor, taste and overall performance of these three chromium enriched Ganoderma lucidum rice pastas were showed 4.2-4.6, and there were no significantly different among these three rice pastas. Chromium enriched Ganoderma lucidum rice pastas may have great potential development in the market.

博士<br>國立中興大學<br>生物科技學研究所<br>103<br>Abstract Background: The aim of this study was to explore the hypoglycemic effects and mechanisms of the Chinese medicine aqueous extract in steroid-induced insulin-resistant (SIIR) rats. One of Xylaria nigripes (XN) is a medicinal fungus that is broadly used in traditional Chinese medi¬cine, another for Gardenia jasminoides Ellis（GJ）is a medicinal herbs, exhibit a hypoglycemic effect by improving insulin secretion and lowering plasma lipids. Methods: In the present study, we fed a water extract of XN and GJ to steroid-induced insulin-resistant (SIIR) rats and observed changes in signaling proteins in order to elucidate the mechanisms of the insulin-sensitizing effect of XN and GJ and evaluate its possibility as an insulin-sensitizing agent. SIIR rats were randomly divided into a control group (i.e., saline) and experimental groups (XN 500 and GJ 200 mg/kg). Results : In summary, the XN extract may have hypoglycemic effects in normal Wistar and SIIR rats that may have a serotonin-related hypoglycemic effect and enhance insulin sensitivity in the SIIR rats. The optimal dose of GJ aqueous extract of 200 mg/kg exerts a PPARγ-activating hypoglycemic effect and improves insulin resistance in SIIR rats. Conclusions: Therefore, they were a potential an insulin-sensitizing agent in type 2 diabetes mellitus with insulin resistance.

碩士<br>國立屏東科技大學<br>生物科技系所<br>104<br>The stem of Cucurbita moschata has been confirmed to contain hypoglycaemic components, but the hypoglycaemic effects of other tissues of the plant are not clear. The purpose of this study is to investigate and compare the hypoglycaemic effects of the stems, the fruits, and the seeds of C. moschata. The rat normal hepatic cell line Clone 9 and the mouse muscle cell line C2C12 were used as models to assay whether the crude extracts of these tissues could promote the glucose consumption of the cells. Consequently, none of the crude extracts showed such an effect in these cells. However, after partitioning by organic solvents, the ethyl acetate (EA) layer of each of the three crude extracts promoted the glucose uptake of the cells, suggesting that the three tissues all contain molecules with insulin-like actities. Further analysis showed that the EA layers of the three tissues all activated AMP- activated protein kinase and increased the tyrosin phosphorylation of IRS-1, suggesting that the insulin-like of the EA layer is mediated by at least two different mechanisms. Subsequently, Clone 9 cells were treated with tumor necrosis factor-alpha (TNF-alpha) to induce insulin resistance, that were treated with the crude extracts of the three tissues. As a result, the crude extracts exhibited insulin sensitizing activites in insulin-resistant cells. Insummary, the stems, fruits, and seeds of C. moschata all contain hypoglycaemic activity, including insulin-like and insulin-sensitizing activities. It is suggested to extract the stems, fruits, or seeds with ethanol, followed by partitioning with acetate, the resulting product can be used manage type 1 and type 2 diabetes.

博士<br>中山醫學大學<br>生化暨生物科技研究所<br>98<br>The objective of this study was to investigate the lipid and fat accumulation-lowering effects of mulberry water extracts (MWEs). To evaluate these effects of MWEs, hamsters were fed with either high fat/cholesterol diets (HFCD) or HFCD supplemented with 0.5, 1 and 2% MWEs for 10 weeks. Plasma total cholesterol (TC) and triglyceride (TG) levels of hamsters fed HFCD with MWEs were significantly reduced by about 44-60% and 14-45%, respectively, as compared to those without MWEs. Similar results were also measured in hepatic TC and TG of hamsters fed HFCD with MWEs. LDLR gene expression and the uptake ability of LDL in HepG2 cells were also upregulated by additions of MWEs. MWEs also decreased the gene expressions of enzymes involved in the TG and TC biosyntheses. Results suggest that hypolipidemic effects of MWEs are via an enhancement of LDLR gene expression and the clearance ability of LDL and a decrease in the lipid biosynthesis. In addition, HFCD-fed groups with MWEs had less perirenal and epididymal fat pads, moreover the weight gain were significantly decreased compared with HFCD-fed groups. These effects probably resulted from MWEs increased the expressions of proteins involved in lipolysis. We further examined the effects of MWEs on protein expressions of the key enzyme of β-oxidation via the HepG2 cell model. Data revealed that the addition in protein expression of PPARα and CPTI in HepG2 cells were detected treatment of 3 and 6 mg/ml of MWEs, respectively, as compared to those without MWEs treatment. The present study reveals for the MWEs could improve hyperinsulinemia and prevent body fat accumulation induced by HFCD. Therefore, MWEs could be used as a natural agent against hyperlipidemia and obesity.

碩士<br>臺北醫學大學<br>醫學科學研究所<br>97<br>The hypercholesterolemia and hyperlipidemia were associated with the high incidence of the formation or development of atherosclerosis and related cardiovascular disorders. Some natural plants have proven they could reduce the concentration of cholesterol or triglycerides in blood serum so that postpone formation or development of cardiovascular disorders. Salvia miltiorrhiza bunge, a traditional Chinese herb which is taken as a blood-quickening, stasis-dispelling medicine, is widely used to improve cardiovascular disorders. Salvianolic acid B is extracted from Salvia miltiorrhiza bunge’s root and exhibites anti-oxidative activity in many assay models. Recent studies demonstrated that salvianolic acid B, curcumin, and the extracts of puerarin and rose had benefit for decreasing the risk factors of cardiovascular disorders. In this study, we used the KDS718 complex to examine the hypolipidemia effects in hamster-fed with high fat diet for 6 or 12 weeks. The KDS718 consists of salvianolic acid B, curcumin, and the crude extract of puerarin and rose, and hamster was free access to powder diet containing various concentrations of powder KDS718. At the 6th weeks and the end, fasting blood was measured the levels of total choleasterol, triglycerides, LDL and VLDL, and the LDL oxidation. In addition, we also examined whether KDS718 had genotoxicity by Ames and mouse lymphoma tk revertant mutagenesis assay, and the micronuclei assay in the mice peripheral blood. Our results suggest that KDS718 can’t improve the hyperlipidemia in hamster-fed with high fat diet, and it has no any genotoxicity. The high fat diet did not induce fatty streak in artery of high fat-fed hamster. KDS718 failed to reduce the lipid levels and induce fatty streak might result from the following reasons. 1. The amounts of active compounds of KDS718 might be not enough to change the lipid metabolism. 2. The hamster selectively ingested powder diet without KDS718 powder. 3. Atherosclerotic lesion might be easy formation with diets containing cholic acid in hamster.

糖尿病是一種以不正常的高血糖為主要特徵的長期性的糖代謝紊亂疾病。二型糖尿病是常見的糖尿病類型，多於九成的糖尿病病人患有此種類型。各種引起糖尿病的病因最終都會導致血糖過高，並且最終會引起有關眼睛，腎臟，神經和血管系統的併發癥。迄今，糖尿病正影響著大約世界6%的人口，而現在患病率依然在逐年增加。在香港，由於高能量的食和缺乏運動，越來越多的老年人和青年人正在遭受著糖尿病的困擾。糖尿病不是一種致命性的疾病，但是如果沒有採取好的治療控制措施，糖尿病最終會引起一些併發癥，這些併發癥最終會使糖尿病患者走向死亡。高血糖癥不僅是糖尿病的主要特徵，而且也是引起各種糖尿病併發癥的重要因素，在二型糖尿病的治療當中，根據各種病理因素，市場上已經研製出了很多西藥來治療糖尿病。然而，它們都有一些副作用的限制。因此，我們需要通過綜合治療和通過新的途徑研製新的製劑來控制血糖水平，保護病人遠離長期併發癥的困擾。如今，腎臟在血糖平衡中的重要角色已經被很好的認知。 在過去的二十年裡， 通過減少血糖在腎臟的重吸收來增加尿液中血糖的排出，從而達到降低體內血糖水平的方法已經被提出并認為是治療糖尿病的一直新的途徑。 在腎臟中，鈉葡萄糖共轉運體2（SGLT 2）主要負責葡萄糖的重吸收，因此，鈉葡萄糖共轉運體2（SGLT 2）抑製劑被認為是一種有潛質的新型的治療糖尿病的製劑。然而，市場上至今沒有成功研製這種製劑。达格列嗪（dapagliflozin），作為一種最有潛質的鈉葡萄糖共轉運體2（SGLT 2）抑製劑，依然處於臨床三期實驗。至今，對具有鈉葡萄糖共轉運體2（SGLT 2）抑製作用的天然產物和傳統中醫藥的信息報導非常少。中醫中藥的治療理念強調整體治療，從此點看來，爲了使糖尿病患者遠離長期的糖尿病併發癥的困擾，中醫中藥可能比西藥更有優勢。<br>因此，本研究的目的是尋找那些具有體外能專門抑制鈉葡萄糖共轉運體2（SGLT 2）並且體內能通過增加尿糖排出來降低血糖水平的抗糖尿天然產物或傳統中藥。從文獻分析中找到了經常用於治療糖尿病的11種中藥和兩種天然產物。<br>試管實驗確立了五味子醇提物和丹皮酚對表達了人的鈉葡萄糖共轉運體2（SGLT 2）基因的COS 7細胞鏈中鈉葡萄糖共轉運體2對¹⁴C-α-甲基- D-葡萄糖苷的吸收作用具有很強的抑制作用。<br>生物活性引導的片段分析確立了五味子醇提物中的活性片段--乙酸乙酯：甲醇（4:6）（F8）片段具有明顯的專門抑制鈉葡萄糖共轉運體2的作用。本實驗也對F8進行了高效液相色譜和液質聯用色譜分析。五味子中三種常見的化合物：五味子甲素，五味子乙素和五味子醇甲存在于F8中，但濃度都很低。試管實驗顯示，這三種常見化合物均無抑制鈉葡萄糖共轉運體2的作用。因此得出結論，這三種常見的五味子化合物不是F8中有效的抑制鈉葡萄糖共轉運體2的活性成份。<br>本實驗也利用動物實驗調查了丹皮酚的抗糖尿作用。糖尿病大鼠被餵食了三個星期的丹皮酚，基礎血糖實驗和尿糖排出實驗均無陽性結果。<br>Diabetes Mellitus (DM) is a chronic disorder of glucose metabolism characterized by abnormally high blood glucose level. Type 2 DM is the common form of diabetes which accounts for more than 90% of all DM cases. All causes of diabetes ultimately lead to hyperglycemia, and it can cause the late complications involving the eyes, kidneys, nerves and blood vessels, which are harmful to health. DM is now affecting about 6% population of the world, and the prevalence is still increasing quickly year by year. In Hong Kong, more and more elderly and youth are suffering from diabetes because of lacking of exercise and high energy diet. DM is not a fatal disease, but if no good action is taken, it can finally cause some kinds of complications, which can lead the patients to the end of their lives. Hyperglycemia is the major characteristics of diabetes, and it is also an important factor which induces all kinds of diabetic complications. In the therapy of type 2 diabetes, a lot of western medicine have been developed in the market according to various pathological causes. However, they have limitations such as existence of side effects. Therefore, combination therapy and development of new agents with novel mechanisms should be required to control the glycemic level and protect the patients from the long-term complications. Nowadays, the significance of the kidney's role in glucose homeostasis is well recognized. Glucose excretion with urine by reducing the renal glucose reabsorption to attenuate the glycemic level has been considered as a new mechanism to treat diabetes since the past two decades. Inhibitors on sodium glucose co-transporters 2 (SGLT 2) which are responsible for the glucose reabsorption in kidney are considered as a kind of new agents that have a potential on the treatment of diabetes. However, there is still no such kind of drug developed in the market, since the most potential one, dapagliflozin, is still on Phase III clinical trial. So far, only few information is found on natural products/traditional Chinese medicines (TCMs) that possess SGLT inhibitory action. Regarding the protection of patients from long-term complications, Chinese medicine which consider the body as a whole, may have advantages over western drugs.<br>Therefore, the aim of this study is to search for anti-diabetic TCM/natural products which specifically inhibit the activity of SGLT2 in vitro and attenuate plasma glucose level in vivo via increasing glucose excretion through urination. From literature review, 11 TCMs and 2 natural products frequently used in treating DM were selected for screening.<br>Using hSGLT 1 and hSGLT 2-expressed COS-7 cell lines as a model, in vitro study demonstrated that Fructus Schisandrae chinensis (ethanolic extract) and paeonol posses the most potent inhibitory effect on SGLT 2 in the in vitro ¹⁴C-α-methyl-D-glucopyranoside (¹⁴C-AMG) uptake assay.<br>The purification of active fraction(s) in ethanolic extract of Schisandrae chinensis fructus was carried out using the bioassay-guided fractionation assay. The ethyl acetate-methanol (4:6) fraction (F8) was selected with significant specific inhibitory effect on SGLT 2. UPLC and LC/MS-MS profiles of F8 were also given in this study. The concentrations of three common compounds of Fructus Shisansrae chinensis: deoxyschisandrin, schisandrin B (γ-schisandrin) and schisandrin were shown very low concentration in F8, the results of uptake assay showed none of these three compounds have inhibitory effects on SGLT 2. It is concluded that these three common compounds in Schisandrae chinensis fructus are not the effective ingredients in F8 which can specifically inhibit SGLT 2.<br>The anti-diabetic effects of paeonol in treating type 2 DM was investigated in animal study. Paeonol (200 and 300 mg/mL) was given to the type 2 diabetic rat model - Zucker Diabetic Fatty (ZDF) rats for three weeks, the results showed no positive effects on the basal glycaemia test and urinary glucose excretion test.<br>Detailed summary in vernacular field only.<br>Detailed summary in vernacular field only.<br>Detailed summary in vernacular field only.<br>Detailed summary in vernacular field only.<br>Detailed summary in vernacular field only.<br>Qu, Yue.<br>Thesis (M.Phil.)--Chinese University of Hong Kong, 2012.<br>Includes bibliographical references (leaves 141-153).<br>Abstracts also in Chinese.<br>TABLE OF CONTENTS<br>ABSTRACT --- p.iv<br>摘要 --- p.vii<br>ACKNOWLEDGEMENT --- p.ix<br>LIST OF ABBREVIATIONS --- p.x<br>LIST OF TABLES --- p.xiii<br>LIST OF FIGURES --- p.xiv<br>TABLE OF CONTENTS --- p.1<br>Chapter CHAPTER 1 --- INTRODUCTION --- p.8<br>Chapter 1.1 --- Definition, diagnosis, classification and epidemiology of Diabetes Mellitus --- p.8<br>Chapter 1.1.1 --- Definition of Diabetes Mellitus --- p.8<br>Chapter 1.1.2 --- Diagnosis of Diabetes Mellitus --- p.8<br>Chapter 1.1.3 --- Classification of Diabetes Mellitus --- p.9<br>Chapter 1.1.4 --- Prevalence of Diabetes Mellitus --- p.11<br>Chapter 1.2 --- Glucose Homeostasis and Diabetes Mellitus --- p.12<br>Chapter 1.2.1 --- General Description --- p.12<br>Chapter 1.2.2 --- Kidney's role in Glucose Homeostasis --- p.14<br>Chapter 1.2.2.1 --- Gluconeogenesis in the Kidney --- p.15<br>Chapter 1.2.2.2 --- Glucose Reabsorption in the Kidney --- p.15<br>Chapter 1.2.2.3 --- Renal glucose transporters --- p.17<br>Chapter 1.2.2.4 --- Disorders with abnormal renal glucose transport --- p.19<br>Chapter 1.3 --- Etiology of Diabetes Mellitus --- p.20<br>Chapter 1.3.1 --- Pancreatic β cell dysfunction --- p.21<br>Chapter 1.3.2 --- Insulin resistance --- p.21<br>Chapter 1.4 --- Diabetic complications --- p.23<br>Chapter 1.5 --- Treatment of type 2 Diabetes Mellitus --- p.25<br>Chapter 1.5.1 --- Conventional therapy of type 2 Diabetes Mellitus --- p.25<br>Chapter 1.5.2 --- New mechanism for the treatment of type 2 Diabetes Mellitus - Inhibition of glucose reabsorption by glucose transporters in Kidney --- p.29<br>Chapter 1.6 --- Traditional Chinese Medicine for Diabetes Mellitus --- p.30<br>Chapter 1.7 --- Project objective --- p.33<br>Chapter CHAPTER 2 --- TRADITIONAL CHINESE HERBAL MATERIALS AND NATURAL PRODUCTS --- p.36<br>Chapter 2.1 --- Materials --- p.36<br>Chapter 2.2 --- General description and anti-diabetic effects of selected herbs/natural products --- p.38<br>Chapter 2.3 --- Extraction Method --- p.45<br>Chapter CHAPTER 3 --- IN VITRO STUDIES OF THE INHIBITORY EFFECT OF SELECTED TRADITIONAL CHINESE HERBS AND NATURAL PRODUCTS ON SODIUM GLUCOSE COTRANSPORTERS (SGLT) --- p.48<br>Chapter 3.1 --- Introduction --- p.48<br>Chapter 3.2 --- Materials --- p.49<br>Chapter 3.3 --- Methods and Methods --- p.52<br>Chapter 3.3.1 --- In vitro model for screening of SGLT inhibitor --- p.52<br>Chapter 3.3.1.1 --- Preparation of hSGLT1 and hSGLT2 Plasmid --- p.52<br>Chapter 3.3.1.2 --- Transient Transfection of SGLT1 or SGLT2 clone --- p.53<br>Chapter 3.3.1.3 --- Detection of mRNA expression level by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) --- p.54<br>Chapter 3.3.1.4 --- Development of SGLT1 or SGLT2 stable cell lines --- p.56<br>Chapter 3.3.1.5 --- Results --- p.56<br>Chapter 3.3.2 --- Cell proliferation assay (MTT assay) --- p.57<br>Chapter 3.3.2.1 --- Methods --- p.57<br>Chapter 3.3.2.2 --- Results --- p.58<br>Chapter 3.3.3 --- Uptake Assay of ¹⁴C-α-methyl-D-glucopyranoside (¹⁴C-AMG) in cultured COS-7 cells expressing SGLT1 or SGLT2 --- p.63<br>Chapter 3.3.3.1 --- Methods --- p.63<br>Chapter 3.3.3.2 --- Screening Results of Effective Chinese Herbs/Natural Products --- p.64<br>Chapter 3.4 --- Discussion --- p.83<br>Chapter CHAPTER 4 --- FRACTIONATION OF SCHISANDRAE CHINENSIS FRUCTUS --- p.86<br>Chapter 4.1 --- Introduction --- p.86<br>Chapter 4.2 --- Organic Extraction of Schisandrae Chinensis Fructus --- p.86<br>Chapter 4.2.1 --- Material and Methods --- p.86<br>Chapter 4.2.2 --- Result --- p.86<br>Chapter 4.3 --- Bioassay-guided Fractionation of Ethanolic Extract of Schisandrae Chinensis Fructus --- p.87<br>Chapter 4.3.1 --- Materials --- p.87<br>Chapter 4.3.2 --- Methods --- p.87<br>Chapter 4.3.2 --- Results --- p.89<br>Chapter 4.4 --- ¹⁴C-α-methyl-D-glucopyranoside (¹⁴C-AMG) Uptake Assay of fractions in cultured COS-7 cells expressing SGLT1 or SGLT2 --- p.92<br>Chapter 4.4.1 --- Methods --- p.92<br>Chapter 4.4.2 --- Results --- p.93<br>Chapter 4.5 --- Characterization of F8 of Schisandrae chinensis fructus using Ultra Performance Liquid Chromatography (UPLC) --- p.98<br>Chapter 4.5.1 --- Introduction --- p.98<br>Chapter 4.5.2 --- Materials and Methods --- p.98<br>Chapter 4.5.3 --- UPLC chromatograms --- p.99<br>Chapter 4.6 --- Characterization of F8 using Liquid Chromatography/Mass Spectrometry-Mass Spectrometry (LC/MS-MS) --- p.101<br>Chapter 4.6.1. --- Materials --- p.101<br>Chapter 4.6.2 --- Methods --- p.102<br>Chapter 4.6.3 --- Results --- p.103<br>Chapter 4.7 --- ¹⁴C-α-methyl-D-glucopyranoside (¹⁴C-AMG) Uptake Assay of three chemical standards in cultured COS-7 cells expressing SGLT1 or SLGT2 --- p.108<br>Chapter 4.7.1 --- Methods --- p.108<br>Chapter 4.7.2 --- Results --- p.108<br>Chapter 4.8 --- Discussion --- p.111<br>Chapter CHAPTER 5 --- IN VIVO STUDIES OF THE ANTI-DIABETIC EFFECT OF SELECTED TRADITIONAL CHINESE HERBS AND NATURAL PRODUCTS IN TYPE 2 DIABETIC RAT MODEL --- p.114<br>Chapter 5.1 --- Introduction --- p.114<br>Chapter 5.1.1 --- Diabetic Animal Models --- p.114<br>Chapter 5.2 --- In vivo Study Tests --- p.117<br>Chapter 5.2.1 --- Introduction --- p.117<br>Chapter 5.2.2 --- Animals --- p.117<br>Chapter 5.2.3 --- Methods --- p.118<br>Chapter 5.2.4 --- Results --- p.120<br>Chapter 5.3 --- Discussion --- p.125<br>Chapter CHAPTER 6 --- GENERAL DISCUSSION --- p.128<br>Chapter 6.1 --- Importance of SGLT --- p.128<br>Chapter 6.2 --- Current developed SGLT 2 Inhibitors --- p.130<br>Chapter 6.3 --- Importance and Treatment of DM by TCMs --- p.132<br>Chapter 6.4 --- Screening and Developing drugs from Traditional Chinese medicinal plants --- p.134<br>Chapter 6.5 --- Limitations and Improvements --- p.136<br>Chapter 6.6 --- Future Works --- p.137<br>Chapter 6.7 --- Conclusions --- p.139<br>REFERENCES --- p.141

Loh Shwu Chun.<br>Thesis (M.Phil.)--Chinese University of Hong Kong, 2005.<br>Includes bibliographical references (leaves [109]-120).<br>Abstracts in English and Chinese; appendix also in Chinese.<br>Acknowledgements --- p.i<br>Abstract --- p.ii<br>Abstract (in Chinese) --- p.iv<br>List of Abbreviations --- p.v<br>List of Tables --- p.vii<br>List of Figures --- p.ix<br>Table of Contents<br>Chapter Chapter One: --- Introduction and Study Objectives<br>Chapter 1. --- Introduction --- p.1<br>Chapter 1.1 --- Definition and diagnostic criteria of the metabolic syndrome --- p.2<br>Chapter 1.2 --- Clinical states of the metabolic syndrome --- p.5<br>Chapter 1.2.1 --- Impaired Glucose Tolerance (IGT) and Impaired Fasting Glucose (IFG) --- p.6<br>Chapter 1.2.2 --- The metabolic syndrome and type 2 diabetes mellitus --- p.7<br>Chapter 1.2.3 --- Dyslipidaemia --- p.8<br>Chapter 1.2.4 --- Hypertension --- p.10<br>Chapter 1.2.5 --- Obesity --- p.11<br>Chapter 1.3 --- Effects of weight loss on the metabolic syndrome --- p.13<br>Chapter 1.4 --- Ethnic differences in the prevalence of the metabolic syndrome --- p.15<br>Chapter 1.5 --- Treatment of the metabolic syndrome --- p.16<br>Chapter 1.6 --- Oral Hypoglycaemic agents and their failure in the metabolic syndrome --- p.17<br>Chapter 1.6.1 --- Sulphonylureas --- p.17<br>Chapter 1.6.2 --- Biguanides --- p.18<br>Chapter 1.6.3 --- Alpha-glucosidase Inhibitors --- p.20<br>Chapter 1.6.4 --- Peroxisome Proliferator-Activated Receptors (PPARs) --- p.21<br>Chapter 1.6.4.1 --- Thiazolinedinediones --- p.22<br>Chapter 1.6.4.1.1 --- Rosiglitazone --- p.24<br>Chapter 1.6.4.1.1.1 --- Mode of Action --- p.24<br>Chapter 1.6.4.1.1.2 --- Adverse events and current status --- p.26<br>Chapter 1.7 --- Orlistat --- p.27<br>Chapter 1.7.1 --- Mode of Action --- p.28<br>Chapter 1.7.2 --- Adverse events and current status --- p.28<br>Chapter 1.7.3 --- Therapeutic Potential in the Metabolic Syndrome --- p.29<br>Chapter 1.8 --- Study Hypothesis --- p.30<br>Chapter 1.9 --- Study Objectives --- p.30<br>Chapter Chapter Two: --- Research Design and Methods<br>Chapter 2 --- Study Protocol --- p.31<br>Chapter 2.1 --- Overall Design --- p.31<br>Chapter 2.1.1 --- Patients Selection Criteria --- p.31<br>Chapter 2.1.1.1 --- Inclusion Criteria --- p.31<br>Chapter 2.1.1.2 --- Exclusion Criteria --- p.33<br>Chapter 2.1.2 --- Recruitment Period --- p.34<br>Chapter 2.1.2.1 --- Screening Period --- p.34<br>Chapter 2.1.2.2 --- Run- In Period (Visit 0) --- p.35<br>Chapter 2.1.2.3 --- Randomisation --- p.35<br>Chapter 2.1.2.4 --- Evaluation Periods (Visit 2 to 4) --- p.37<br>Chapter 2.2 --- Investigations --- p.37<br>Chapter 2.2.1 --- Oral Glucose Tolerance Test (OGTT) --- p.38<br>Chapter 2.2.2 --- Anthropometric measurements --- p.38<br>Chapter 2.3 --- Analytical Methods --- p.39<br>Chapter 2.3.1 --- Determinations of insulin levels in plasma samples --- p.39<br>Chapter 2.3.1.1 --- Principle of the Insulin assay --- p.40<br>Chapter 2.3.2 --- Determinations of glucose concentrations in samples --- p.42<br>Chapter 2.3.2.1. --- Principle of the glucose assay --- p.42<br>Chapter 2.4 --- Calculations --- p.43<br>Chapter 2.4.1 --- Insulin (hepatic) sensitivity (HOMA) --- p.43<br>Chapter 2.4.2 --- Area Under the Curves --- p.44<br>Chapter 2.4.3 --- Sample Size Calculations --- p.45<br>Chapter 2.5 --- Statistical Analysis --- p.46<br>Chapter Chapter Three: --- Results<br>Chapter 3.1 --- Study Population --- p.48<br>Chapter 3.2 --- Randomisation --- p.49<br>Chapter 3.3 --- Study Results --- p.50<br>Chapter 3.3.1 --- Indices of Glycaemic Control --- p.54<br>Chapter 3.3.1.1 --- HbAlc --- p.54<br>Chapter 3.3.1.2 --- Fasting Plasma Glucose --- p.58<br>Chapter 3.3.1.3 --- Fasting Insulin --- p.58<br>Chapter 3.3.1.4 --- 75g Oral Glucose Tolerance Test --- p.59<br>Chapter 3.3.1.4.1 --- Glucose --- p.59<br>Chapter 3.3.1.4.1.1 --- 2hr-Glucose --- p.61<br>Chapter 3.3.1.4.1.2 --- GlucoseAuc --- p.62<br>Chapter 3.3.1.4.2 --- Insulin --- p.63<br>Chapter 3.3.1.4.2.1 --- 2-hr insulin --- p.63<br>Chapter 3.3.1.4.2.2 --- InsulinAuc --- p.65<br>Chapter 3.3.1.5 --- HOMA score --- p.67<br>Chapter 3.3.2 --- Clinical Determinants --- p.69<br>Chapter 3.3.2.1 --- Lipid Profiles --- p.69<br>Chapter 3.3.2.1.1. --- Total Cholesterol --- p.69<br>Chapter 3.3.2.1.2 --- HDL-Cholesterol --- p.70<br>Chapter 3.3.2.1.3 --- LDL-Cholesterol --- p.71<br>Chapter 3.3.2.1.4 --- Triglycerides --- p.72<br>Chapter 3.3.2.2 --- Anthropometric Evaluations --- p.74<br>Chapter 3.3.2.2.1 --- Body Weight --- p.74<br>Chapter 3.3.2.2.2 --- Waist Circumference Difference --- p.75<br>Chapter 3.3.2.2.3 --- Hip --- p.76<br>Chapter 3.3.2.2.4 --- Body Fat --- p.78<br>Chapter 3.3.2.2.5 --- BMI --- p.78<br>Chapter 3.3.2.3 --- Blood Pressure --- p.79<br>Chapter 3.3.2.4 --- RCCA and LCCA --- p.79<br>Chapter 3.3.2.5 --- Other outstanding measurements --- p.82<br>Chapter 3.4 --- Side Effects experienced --- p.82<br>Chapter Chapter Four: --- Discussion and Conclusion<br>Chapter 4.1 --- Summary of the results --- p.83<br>Chapter 4.1.1 --- Effects of Diet and Lifestyle Changes --- p.83<br>Chapter 4.1.2 --- Effects of Orlistat --- p.84<br>Chapter 4.1.3 --- Effects of Rosiglitazone --- p.35<br>Chapter 4.2 --- Implications for therapy --- p.86<br>Chapter 4.2.1 --- Management of metabolic syndrome --- p.87<br>Chapter 4.2.2 --- Early Diagnosis --- p.88<br>Chapter 4.2.3 --- Lifestyle Modification --- p.89<br>Chapter 4.2.4 --- Pharmacological Targets --- p.92<br>Chapter 4.2.4.1 --- Statins --- p.92<br>Chapter 4.2.4.2 --- Fibrates --- p.93<br>Chapter 4.2.4.3 --- ACE Inhibitors --- p.93<br>Chapter 4.2.4.4 --- Thiazolidinediones --- p.94<br>Chapter 4.2.4.4.1 --- Economic Evaluations of Thiazolidinediones --- p.97<br>Chapter 4.2.4.5 --- Orlistat --- p.98<br>Chapter 4.2.4.5.1 --- Economic Evaluations of Orlistat --- p.102<br>Chapter 4.3 --- Limitations of the study --- p.104<br>Chapter 4.3.1 --- Small sample size --- p.104<br>Chapter 4.3.2 --- Short period of study --- p.105<br>Chapter 4.3.3 --- Adherence to lifestyle modifications --- p.105<br>Chapter 4.3.4 --- Analytical assays --- p.106<br>Chapter 4.3.5 --- Follow up end of study --- p.106<br>Chapter 4.3.6 --- Ultrasound measurement of the common carotid arteries --- p.106<br>Chapter 4.3.7 --- Availability of thiazolinediones --- p.107<br>Chapter 4.4 --- Conclusion and Implications for future studies --- p.107<br>References --- p.110<br>Appendix I --- p.121<br>Appendix II --- p.122<br>Appendix III --- p.125

AIMS/HYPOTHESIS: We previously demonstrated that animals fed a high-fat (HF) diet for 10 weeks developed insulin resistance and behavioural inflexibility. We hypothesised that intervention with metformin would diminish the HF-feeding-evoked cognitive deficit by improving insulin sensitivity. METHODS: Rats were trained in an operant-based matching and non-matching to position task (MTP/NMTP). Animals received an HF (45% of kJ as lard; n = 24), standard chow (SC; n = 16), HF + metformin (144 mg/kg in diet; n = 20) or SC + metformin (144 mg/kg in diet; n = 16) diet for 10 weeks before retesting. Body weight and plasma glucose, insulin and leptin were measured. Protein lysates from various brain areas were analysed for alterations in intracellular signalling or production of synaptic proteins. RESULTS: HF-fed animals developed insulin resistance and an impairment in switching task contingency from matching to non-matching paradigm. Metformin attenuated the insulin resistance and weight gain associated with HF feeding, but had no effect on performance in either MTP or NMTP tasks. No major alteration in proteins associated with insulin signalling or synaptic function was detected in response to HF diet in the hypothalamus, hippocampus, striatum or cortex. CONCLUSIONS/INTERPRETATION: Metformin prevented the metabolic but not cognitive alterations associated with HF feeding. The HF diet protocol did not change basal insulin signalling in the brain, suggesting that the brain did not develop insulin resistance. These findings indicate that HF diet has deleterious effects on neuronal function over and above those related to insulin resistance and suggest that weight loss may not be sufficient to reverse some damaging effects of poor diet.

Hyperphosphorylated tau plays an important role in the formation of neurofibrillary tangles in brains of patients with Alzheimer's disease (AD) and related tauopathies and is a crucial factor in the pathogenesis of these disorders. Though diverse kinases have been implicated in tau phosphorylation, protein phosphatase 2A (PP2A) seems to be the major tau phosphatase. Using murine primary neurons from wild-type and human tau transgenic mice, we show that the antidiabetic drug metformin induces PP2A activity and reduces tau phosphorylation at PP2A-dependent epitopes in vitro and in vivo. This tau dephosphorylating potency can be blocked entirely by the PP2A inhibitors okadaic acid and fostriecin, confirming that PP2A is an important mediator of the observed effects. Surprisingly, metformin effects on PP2A activity and tau phosphorylation seem to be independent of AMPK activation, because in our experiments (i) metformin induces PP2A activity before and at lower levels than AMPK activity and (ii) the AMPK activator AICAR does not influence the phosphorylation of tau at the sites analyzed. Affinity chromatography and immunoprecipitation experiments together with PP2A activity assays indicate that metformin interferes with the association of the catalytic subunit of PP2A (PP2Ac) to the so-called MID1-alpha4 protein complex, which regulates the degradation of PP2Ac and thereby influences PP2A activity. In summary, our data suggest a potential beneficial role of biguanides such as metformin in the prophylaxis and/or therapy of AD.