葛根素对实验性糖尿病胰岛素抵抗作用及其机制的研究
摘要
目的:观察葛根素(Pue)对糖尿病胰岛素抵抗(IR)的作用,及其作用机制。方法:(1)以链脲菌素(STZ)诱导糖尿病(DM)小鼠模型和以氢化可的松琥珀酸钠(HCSS)诱导IR小鼠模型,观察Pue对正常小鼠、DM模型小鼠及IR模型小鼠血糖、血清胰岛素含量及糖耐量的影响。以小剂量STZ和高脂饲料喂养诱导IR大鼠模型,通过糖耐量实验(IPGTT)和高血浆胰岛素-正糖钳夹实验(HECT)来考察Pue对IR大鼠胰岛素敏感性,并检测大鼠血浆中胰岛素(Insulin)、血浆甘油三酯(TG)、胆固醇(TC)、肿瘤坏死因子a(TNFa)、瘦素(Leptin)的含量,以探讨Pue防治糖尿病胰岛素抵抗的作用机制。(2)体外培养BRL大鼠肝细胞,用高胰岛素诱导其形成IR细胞模型,通过葡萄糖消耗实验和对细胞培养液中NO、NOS的检测,观察Pue对肝细胞的葡萄糖消耗和肝细胞NO合成的影响。采用逆转录-聚合酶链反应方法,观察Pue对肝细胞胰岛素降解酶(IDE)基因表达的影响。
结果:Pue对正常小鼠空腹血糖无明显影响,可明显降低STZ诱导的糖尿病小鼠的血糖和血清胰岛素含量,使胰岛素敏感性指数显著升高,对HCSS诱导的IR也有明显的改善;Pue可改善小剂量STZ和高脂饲料喂养诱导IR大鼠的葡萄糖耐量,增加单位胰岛素对葡萄糖的转化率,降低其血糖、血脂及血中Insulin、TNFa、Leptin的含量;Pue可增加高胰岛素环境下大鼠肝细胞的葡萄糖消耗量,调节其NOS的活性和NO的合成,降低其胰岛素降解酶的基因表达。结论:Pue可明显改善实验性糖尿病胰岛素抵抗,其作用机制主要是:一方面通过降低血糖和血清胰岛素水平来保护胰岛β细胞及肝细胞的功能,另一方面通过降低血浆TG、TC、TNFa、Leptin水平,调节肝细胞NOS的活性和NO的合成,降低IDE的基因表达,改善肝脏及外周组织对胰岛素的敏感性而实现的。
Objective: To investigate the effect of Puerarin (Pue) on the Experimental diabetes insulin resistance (IR) and its mechanism. Methods: IR mice model induced Streptozotocin (STZ) and Hydrocortisone Sodium Succinate (HCSS). Study the influence of Pue on fasting blood sugar, fasting serum insulin and sugar tolerance on normal mice and model mice. Inject vein small dosage STZ and high- fat feed induced IR rat model. Study the effect of Pue on insulin sensitivity on IR rat by sugar tolerance test and hyperinsulinemic euglycenic clamp technique (HECT). Probe into the mechanism of Pue by detecting the contents of plasma insulin, triglyceride (TG), total cholesterol (TC), tumor necrosis factor α (TNF- α )and Leptin (Lep). Rat hepatic cell (BRL) were incubated for 16 hour in high insulin condition to induce IR cell model. Study the effect of Pue on the glucose consumption and NO synthesis of hepatic cell. Results: Pue has little influence on fasting blood glucose of normal mice, but can lower blood sugar and serum insulin of diabetic mice by STZ induced and increase insulin sensitivity index significantly as well as improve IR by HCSS induced significantly. Pue can improve the glucose tolerance and glucose transformation rate of insulin, lower blood sugar and blood fat(TG,TC), decrease the contents plasma insulin, TNF- α and Leptin of IR rat induced by small dosage STZ and high-fat feet. Also Pue can increase glucose consumption of rat hepatic cell in high insulin condition through regulating NO synthesis as well as reducing IDE expression. Conclusion: Pue can improve IR of the Experimental diabetes by decrease blood sugar and serum insulin level to protect β cell and hepatic cell, as well as lower TG, TC, TNF- α , Leptin and adjust NO synthesis, reduce the expression of IDE gene to improve the insulin sensitivity of hepatic and periphery tissue.
结果:Pue对正常小鼠空腹血糖无明显影响,可明显降低STZ诱导的糖尿病小鼠的血糖和血清胰岛素含量,使胰岛素敏感性指数显著升高,对HCSS诱导的IR也有明显的改善;Pue可改善小剂量STZ和高脂饲料喂养诱导IR大鼠的葡萄糖耐量,增加单位胰岛素对葡萄糖的转化率,降低其血糖、血脂及血中Insulin、TNFa、Leptin的含量;Pue可增加高胰岛素环境下大鼠肝细胞的葡萄糖消耗量,调节其NOS的活性和NO的合成,降低其胰岛素降解酶的基因表达。结论:Pue可明显改善实验性糖尿病胰岛素抵抗,其作用机制主要是:一方面通过降低血糖和血清胰岛素水平来保护胰岛β细胞及肝细胞的功能,另一方面通过降低血浆TG、TC、TNFa、Leptin水平,调节肝细胞NOS的活性和NO的合成,降低IDE的基因表达,改善肝脏及外周组织对胰岛素的敏感性而实现的。
Objective: To investigate the effect of Puerarin (Pue) on the Experimental diabetes insulin resistance (IR) and its mechanism. Methods: IR mice model induced Streptozotocin (STZ) and Hydrocortisone Sodium Succinate (HCSS). Study the influence of Pue on fasting blood sugar, fasting serum insulin and sugar tolerance on normal mice and model mice. Inject vein small dosage STZ and high- fat feed induced IR rat model. Study the effect of Pue on insulin sensitivity on IR rat by sugar tolerance test and hyperinsulinemic euglycenic clamp technique (HECT). Probe into the mechanism of Pue by detecting the contents of plasma insulin, triglyceride (TG), total cholesterol (TC), tumor necrosis factor α (TNF- α )and Leptin (Lep). Rat hepatic cell (BRL) were incubated for 16 hour in high insulin condition to induce IR cell model. Study the effect of Pue on the glucose consumption and NO synthesis of hepatic cell. Results: Pue has little influence on fasting blood glucose of normal mice, but can lower blood sugar and serum insulin of diabetic mice by STZ induced and increase insulin sensitivity index significantly as well as improve IR by HCSS induced significantly. Pue can improve the glucose tolerance and glucose transformation rate of insulin, lower blood sugar and blood fat(TG,TC), decrease the contents plasma insulin, TNF- α and Leptin of IR rat induced by small dosage STZ and high-fat feet. Also Pue can increase glucose consumption of rat hepatic cell in high insulin condition through regulating NO synthesis as well as reducing IDE expression. Conclusion: Pue can improve IR of the Experimental diabetes by decrease blood sugar and serum insulin level to protect β cell and hepatic cell, as well as lower TG, TC, TNF- α , Leptin and adjust NO synthesis, reduce the expression of IDE gene to improve the insulin sensitivity of hepatic and periphery tissue.
引文
1、Kruszynska YT, Yu JG, O lefsky JM, et al. Effects of troglitazone on blood concentrations of plasminogen activator inhibitor in patients with type 2 diabetes and in lean and obese normal subjects. Diabetes, 2000, 49: 633-639.
2、Reaven G. Diabetes, 1988, 37: 1595-1607.
3、Haffner SM. Epidemiology of insulin resistance and its relation to coronary artery disease. American Journal of Cardiology. 1999, 84(1A): 11-14.
4、Reaven GM. Insulin resistance: a chicken that has come to roost. Annals of the New York Academy of Sciences. 1999, 892: 45-57.
5、Tripathy D, Carlsson M, Almgren P, Isomaa B, Taskinen MR, Tuomi T, Groop LC. Insulin secretion and insulin sensitivity in relation to glucose tolerance: lessons from the Botnia Study. Diabetes, 2000, 49(6):975-980.
6、 Groop L, Orho-Melander M. The dysmetabolic syndrome. Journal of Internal Medicine, 2001, 250(2): 105-120.
7、Jia Weiping, Xiang Kunsan, Chen Lei, Lu Junxi, Bao Yuqian, Wu Yuanmin, Jiang Sunyin. A comparison of the application of two working definitions of metabolic syndrome in Chinese population. Natl Med J China, 2004, 84(7): 534-538.
8、Liu Xing, Wang Yalou, et al. Present states and developmental trends of therapeutic drugs for type Ⅱ diabetes. Journal of Pharmaceutical Practice, 2004, 21(1): 1-4.
9、Cai Zhongde, Wu Jie, Zhu Huirong. The General Situation of the study of Chinese Traditional Medicine Curing Diabetes Mellitus. Chinese Journal of Experimental Traditional Medical Formulae. 2002, 8 (6): 56-59.
10、Xie Qingdong. The Applications of Puerarin in Heart and Brain' s angiopathy. Medicine and Materia Medicine Research. 2002, 13 (2): 96-97.
11、Shen Zuofang, Xie Mingzhi. Effects of puerarin and compound aspirin on blood sugar. Medicine Transaction. 1985, 20 (11): 863-865.
12、Pan Hongping. Study progress of Radix puerarin Taurine and their compound preparation in decrease of Blood-glucose. Chinese Traditional Patent Medicine. 2002, 24 (3): 221-224.
13、Liu Xiaojun, Liang Hongyu, Li Guiming. The investigation of the effect of puerarin's injection in curing DN in early state and its improving living quality. Modern Rehabilitation. 2001, 5(6):135-137.
14、Kang Shengqun, Zhao shujian, Zhao Wenhui, et al. Clinic observe on the effect of puerarin on diabetes mellitus nephropathy. Clinic Assemble. 2002, 17 (8): 440-441.
15、Li ChenZhong, Zhang Su Hua, Shu ChangDa. Expression of insulin degrading enzyme in insulin resistance cell model. Chin J Endocrinol Metab, 2000, 16(5): 302-305.
16、Liu Y L, Chen S H, Chen X B. Effects of puerarin on insulin-sensitivity in type 2 diabetic sufferer. Liaoning Journal of Utility Diabetic, 2000, 8(1): 26.
17、Shong ChunYu, Bi HuiMin, Effects of puerarin on plasma membrane GLUT4 content in skeletal muscle from insulin resistant Sprague-dawley rats under insulin stimulation. China Journal of Chinese Materia Medica, 2004, 29(2): 172-175.
18、Wu Jian, Lei Moxian. New consideration on the relation of insulin resistance and type 2 diabetic. Medicine and Philosophy, 2004, 25(2): 4-5.
19、Mao Caipin, Gu Zhenlun. Experimental studies of effects of puerarin on hemorheology in diabetic rats. Chinese Journal of Hemorheology. 2003, 13(4): 322-324.
20、Greene DA. Acute and chronic complications of diabetes mellitus in older patients. American Journal of medicine. 1986, 80(5A): 39-53.
21、Chen JB, Xu JL, Jiang H, et al. The effects of puerarin on endothelin and blood platelet in patients suffering from coronary heart disease. Zhejiang J Tradit Chin Med (in Chinese). 1999, 34(5): 216-217.
22、Lu XR, Gao E, Xiu LZ, et al. Blocking effect of puerarin on β-adrenoceptor of isolated organs and whole animal. Acta Pharmacol Sin. 1986, 7(6): 537-539.
23、Zhang HY, Xu YB, Wang XJ, et al. Effects of puerarin on function of platelet and microalbuminuria in diabetes mellitus. Chin J New Drugs Clin Rem. 1999, 18(5): 291-293
24、Wang Jian, Chen Min, Cao Yi, et al. Effect of Kakonein and Gingko Flavone on Protein Nonenzymatic. Parmacology and Clinical Applications of Chinese Traditional Medicine. 2000, 16(1):13-14.
25、Zhang J T. Modern Methodology in Pharmacological Experiment. Beijin: Peking Medical College and Peking Union Medical College Publishing House, 1998: 981-982.
26、Li G W, Pan X R, Stephen L, et al. A new insulin-sensitivity index for the population-based study. Chin J Intern Med. 1993, 32(10): 656-660.
27、Moller DE, Flier JS. Insulin resistance mechanism, syndromes, and impli cations. N Eng J med, 1991, 325(13): 928-948.
28、Storlien LH, James DE, Burleigh KM, et al. Fat feeding causes widespread in vivo insulin resistance, decreased energy expenditure, and obesity in rats . AmJ Physiol, 1986, 251: E576-583.
29、Ding Shiying, Shen Zhufang, Chen Yueteng, et al. Using euglycemic clamp technique estimate two type of insulin-resistance animal model. Chinses Diabetes magazine, 2001, 9 (5): 286-289.
30、Li Chenzhong, Zhang Suhua, Shu Changda, et al. High fat meal induce insulin-resistance rats. Basic Medical Sciences and Clinics, 2000, 20(3): 93-95.
31、Chen Qi. Pharmacology of Trasitional Chinese Medicine Experiment Method, Dumb Millions Sanitation Publishing Company, 1993: 719.
32、Yin Jun, Chen MingDao, Zhou LiBing, et al. Effects of long-term high-fat diet on the occurrence of diabetes in rats. Journal of Chinese Physician, 2001, 6(1):47-49.
33、Chen Lin, Bai Jie. Influence of Fibrin, Chromium and Zinc on glucose tolerance of rats, Liaoning Journal of Utility Diabetic. 2001, 9(1):28-29.
34、Sakamoto S, Minami K, Niwa Y, et al. Effect of exercise training and food restriction on endothelium dependent Re2 laxation in the otsuka Long Evans Tokushima fatty rat, a model of spontaneous NIDDM. Diabetes, 1998, 47(1): 82.
35、Miles PDG, Higo K, Romeo OM, et al. Troglitazone prevents hyperglycemia induced but not glucosamine induced insulin resistance. Diabetes, 1998, 47(3): 395.
36、Du Jian, Zhao YuYan, Xie Hui, Zhao Jin Establishment and evaluation of animal model of insulin resistance with high fatty diet. J Chin Med Univ, 2002, 31(5):343-346.
37、Hevener AL, Reichart D, Olefsky J, et al. Thiazolidinedione treatment prevents free fatty acid-induced insulin resistance in male wister rats. Diabetes, 2001, 50: 2316-2322.
38、Mannucci E, Bardini G, Ognibene et al. Comparison between insulin sensitivity indexes in obese patients. Diabetes Care, 2000, 23(7): 1042-1043.
39、Bokemark L, Froden A, Attvall S, et al. The euglycemic hyperinsulinmic clamp examination: variability and reproducibility. Scand J Clin Lab Invest, 2000, 60(1):27-36.
40、Yang YongMei, Zhang NanYan. Effect of free fatty acids on diabetes fat toxicity. Chinese Journal of Misdiagnostics, 2003, 3(1):48.
41、Hui, Zhang Yah. High fatty feed induce insulin resistance animal model and estimate. Chinese Journal of Misdiagnostics, 2003, 3(10): 1531-1532.
42、Goetze S, Blaschke F, Stawowy P, et al. Biochem Biophys ResCommun, 2001, 287(3): 662-670.
43、Sheu WHH, lee WJ, Chang RL, et al. Clin Exp Hypertens, 2000, 22(6): 595-606.
44、Fruhbeck C, Gomez-Ambrosi J, Muruzabal FJ, et al. The adipocyte, a model for integration of endocrine and metabolic signaling in energy metabolism regulation. Am J Physiol Endocrinol Metab, 2001, 280: 827.
45、Cohen PG. Aromatase, adiposity, aging and disease. The hypogonadal-metabolic-atherogenic-disease and aging connection. Med Hypotheses, 2001, 56(6):702-708.
46、Jung M, et al. Infulence of varied PH cultured media on nitric oxide synthesis of hepatocytes. Chin J Dig, 2000, 20(1)26-29.
47、D Ambrosio SM, Gibson-D Ambrosio RE, Brady T, et al. Mechanisms of nitric oxide-induced cytotoxicity in Normal human hepatocytes. Environ Mol Mutagen, 2001, 37(1):46-54.
48、Liu GuoQing. Study headway on NO and diabetes syndrome. J Overseas Physic Incretion Fascicule, 1999, 19(6):257-259.
49、Li ChenZhong, Zhang SuHua, Su ChangDa, et al. Establishing and identifing insulin-resistance cell line. Acta Universitatis Scientiae Medicine ChongQing, 1998, 23:335-337.
50、Li ChangGui, Ning Guang, Chen JiaLun. Establishing and identifying insulin-resistant HepG2 cell line. Chinese Journal of Diabetes, 1999, 7(4):198-200.
51、Ying Jun, Hu RenMing, Chen MingDao, et al. Compare the effect of glucose disappeared on metformin and berberine on HepG2 cell. Chin J Endocrinol Metab, 2002, 18(6):488-489.
52、Hansen MB, Nielsen SE, Berg K. Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J Immunol Method, 1989, 119:203-210.
53、 Wang XiaoYun, Liao KunLing, Lu Wei, et al. The relationship between NO and IR of DM (Type Ⅱ). Guizhou Medical Journal, 2001, 25(1):18-19.
54、 Kuo WL, Gehm BD, Rosner MR, et al. Regulation of insulin degradation: expression of an evolutionarily conserved insulin degrading enzyme increase degradation by an intracellular pathway. Mol Endocrinol, 1991, 5:1467-1476.
55、 Authier F, Ponsner BF, Bergeron JJ. Insulin-degrading enzyme. Clin Invest Med, 1996, 19(2) :149-152.
56、 Michihiro Fukushima, Masuo Nakano, Yasuko Morii, et al. Hepatic LDL Receptor mRNA in Rats Is Increased by Dietary Mushroom (Agaricus bisporus) Fiber and Sugar Beet Fiber. Journal of Nutrition, 2000, 130:2151-2156.
57、 Caro J F, Amutrvda J M. Glucocorticoid induced insulin resistance of man. J Clin Invest, 1982, 69:866.
58、 Dgawa A. Roles of insulin resistance and β -cell dysfuntion in Dexamethasone-indncel diabetes. J Clin Invest, 1992, 90:497.
59、 Campbell K L, Latimer KS. Transient diabetes mellitus associated with prednisone therapy in dog. J Am Vet Med Assoc, 1984,185:299.
60、 Moller DE, Flier JS. Insulin resistance mechanism, syndromes, and implications. N Eng J med, 1991,325(13): 928-948.
61、 Kragen EW, Clark PW, Jenkins AB, et al. Development of muscie insulin resistance in high-fat-fed rats. Diabetes, 1991, 40:1397-1403.
62、 Storlien LH, Higgins JA , Thomas TC , et al. Diet composition and insulin action in animal models. British Journal of Nutrition, 2000, 83 (Suppll): S85-S90.
63、 Ye JM, Doyle PJ, Lglesias MA, et al. Peroxisome proliferator activated receptor (PPAR) alpha activation lowers muscle lipids and improves insulin sensitivity in high fat-fed rats: comparison with PPAR-gamma activation. Diabetes, 2001, 50(2):411-417.
64、Storlien LH, Oakes ND, Pan DA, et al. Syndrome of insulin resistan in the rats: inducement by diet and amelioration with Benfliorex. Diabetes, 1993, 42:457-462.
65、Yang GangYi, Peter Chung, Guenther Boden. Changes of glucose metabolism in insulin clamp studies in the non-anaesthesia rats. Basic Medical Sciences and Clinics. 2003, 23(3): 341-344.
66、Kraegen EW, James DE, Bennett SP, et al. In vivo insulinsensitivity in the rat determined by euglycemic clamp. Am J Physiol, 1983, 245 (Endocriol. Metab, 8): E1-7.
67、Yang GangYi, Li Ling, Peter Chung, Gangther Boden. Effects of phosphodiesterase inhibitor-induced insulin resistance on glucose and lipid metabolism in rats. Chinese Journal of Pathophysiology, 2003, 19(7): 875-880.
68、Lu HuiLing, Wang HongWei, Ling HanHua, Zhang MuXun. Relation of resistin with blood glucose, blood fattiness and insulin resistance on diabetes sufferer. Clin J Intern Med, 2004, 43(1):53-54.
69、Liang Zhen, Cheng Hua. Lipotoxicity and type 2 diabetes. J Overseas Physic Medicine Fascicule, 2004, 31(2):50-53.
70、Lupi R, Del Guerra S, Fierabracci V, et al. Lipotoxicity in human pancreatic islets and the protective effect of metformin. Diabetes, 2002, 51 Suppll:S134-137.
71、Mc Garry JD. Banting lecture 2001: dysregulation of fatty acid metabolism in the etiology of type 2 diabetes. Diabetes, 2002, 51: 7-18.
72、Cheung C, Clifton D, Steiner R. Proopiomelanocortin neurons are direct targets for leptin in the hypothalamus. Endocrinology, 1997, 138: 4489-4492.
73、Kieffer TJ, Heller RS, Leech CA, et al. Diabetes, 1997, 46: 1087-1093.
74、Stejskal D, Jedelsky L, Bartek J, et al. Vnitr Lek, 1998, 44:361-365.
75、Cao Xiaopei, Ming Wenyu, Cheng Hua, et al. Effects of leptin on glucose oxidation and glucokinase gene expression in cultured liver cells. Chin J Endocrinol Metab, 1999, 15(3): 131-133.
76、Kennedy A, Gettys TW, Watson P, et al. J Clin Endocrinol Metab, 1997, 82: 1293-1300.
77、L fgren P, Van Harmelen V, Reynisdottir S, et al. Diabetes, 2000, 49 (5): 688-692.
78、Fasshauer M, Klein J, Neumann S, et al. Biochem Biophys Res Commun, 2001, 288 (4): 1027-1031.
79、Nicaud V, Raoux S, Poirier O, et al. Atherosclerosis, 2002, 161 (2): 317-325.
80、Ronnemaa T, Pulkki K, Kaprio J, et al. J Clin Endocrinol Metab, 2000, 85 (1): 2728-2732.
81、Liu S, Spelleken M, R brig K, et al. Diabetes, 1998, 47(4):515-522.
82、Goetze S, Blaschke F, Stawowy P, et al. Biochem Biophys Res Commun, 2001, 287 (3): 662-670.
83、Sheu WH, Hlee WJ, Chang RL, et al. Clin Exp Hypertens, 2000, 22(6):595-606.
84、Nisoli E, Carruba MO, Tonello C, et al. Diabetes, 2000, 49(3):319-325.
85、Prins JB, Niesler CU, Winterford CM, et al. Diabetes, 1997, 46 (12): 1939-1944.
86、Rachael L, Fawcett, Angela S, et al. J Clin Endocrinol Metab, 2000, 85 (2): 530-535.
87、Zhu XiXing. β-cell dysfunction and insulin resistance. J Overseas Physic Incretion Fascicule, 2001, 21(4):169-176.
88、Weir CC, Laybutt DR, Kaneto H, et al. β-cell adaptation and decompensation during the progression of diabetes. Diabetes, 2001, 50(suppll): S154-159.
89、Shen Cuzhou. Insulin recept and insulin signal conduct access with insulin resistance. A cad J Sec Mil Med University, 2001, 22(11): 1062-1063.
90、Heck DE. NO, RSNO, ONOO-, NO-NOO, Nox-dynamic regulation of oxidant scavenging, nitric oxide stores, and cyclic GMP-independent cell signaling. Antioxid Redox Signal, 2001, 3(2): 249-260.
91、Molecular Clone Experiment Enchieidion. J Sambluker, et al. Jin DongYan, Li MengFeng translated. Beijing Science Publishing Company. 1996, 683-688.
92、Zeng G, Ouon MJ. Insulin-stimulated production of nitric oxide is inhibited by wortmannin. Direct measurement in vascular endothelial cells J Clin Invest, 1996; 98(4):894-898.
93、Zhang Hui, Chen XiangFang, Ye FuLin, Liu ZhiNian, et al. Effects of insulin on nitric oxide synthase (NOS) activity and the expression of inducible NOS mRNA in calf aortic endothelial cells. Chin J Endocrinol Metab, 2003, 19(6): 483-485.
94、Stuhlinger MC, Abbasi F, Chu JW, et al. Relationship between insulin resistance and an endogenous nitric oxide synthase inhibitor. JAMA, 2002, 287: 1420-1426.
2、Reaven G. Diabetes, 1988, 37: 1595-1607.
3、Haffner SM. Epidemiology of insulin resistance and its relation to coronary artery disease. American Journal of Cardiology. 1999, 84(1A): 11-14.
4、Reaven GM. Insulin resistance: a chicken that has come to roost. Annals of the New York Academy of Sciences. 1999, 892: 45-57.
5、Tripathy D, Carlsson M, Almgren P, Isomaa B, Taskinen MR, Tuomi T, Groop LC. Insulin secretion and insulin sensitivity in relation to glucose tolerance: lessons from the Botnia Study. Diabetes, 2000, 49(6):975-980.
6、 Groop L, Orho-Melander M. The dysmetabolic syndrome. Journal of Internal Medicine, 2001, 250(2): 105-120.
7、Jia Weiping, Xiang Kunsan, Chen Lei, Lu Junxi, Bao Yuqian, Wu Yuanmin, Jiang Sunyin. A comparison of the application of two working definitions of metabolic syndrome in Chinese population. Natl Med J China, 2004, 84(7): 534-538.
8、Liu Xing, Wang Yalou, et al. Present states and developmental trends of therapeutic drugs for type Ⅱ diabetes. Journal of Pharmaceutical Practice, 2004, 21(1): 1-4.
9、Cai Zhongde, Wu Jie, Zhu Huirong. The General Situation of the study of Chinese Traditional Medicine Curing Diabetes Mellitus. Chinese Journal of Experimental Traditional Medical Formulae. 2002, 8 (6): 56-59.
10、Xie Qingdong. The Applications of Puerarin in Heart and Brain' s angiopathy. Medicine and Materia Medicine Research. 2002, 13 (2): 96-97.
11、Shen Zuofang, Xie Mingzhi. Effects of puerarin and compound aspirin on blood sugar. Medicine Transaction. 1985, 20 (11): 863-865.
12、Pan Hongping. Study progress of Radix puerarin Taurine and their compound preparation in decrease of Blood-glucose. Chinese Traditional Patent Medicine. 2002, 24 (3): 221-224.
13、Liu Xiaojun, Liang Hongyu, Li Guiming. The investigation of the effect of puerarin's injection in curing DN in early state and its improving living quality. Modern Rehabilitation. 2001, 5(6):135-137.
14、Kang Shengqun, Zhao shujian, Zhao Wenhui, et al. Clinic observe on the effect of puerarin on diabetes mellitus nephropathy. Clinic Assemble. 2002, 17 (8): 440-441.
15、Li ChenZhong, Zhang Su Hua, Shu ChangDa. Expression of insulin degrading enzyme in insulin resistance cell model. Chin J Endocrinol Metab, 2000, 16(5): 302-305.
16、Liu Y L, Chen S H, Chen X B. Effects of puerarin on insulin-sensitivity in type 2 diabetic sufferer. Liaoning Journal of Utility Diabetic, 2000, 8(1): 26.
17、Shong ChunYu, Bi HuiMin, Effects of puerarin on plasma membrane GLUT4 content in skeletal muscle from insulin resistant Sprague-dawley rats under insulin stimulation. China Journal of Chinese Materia Medica, 2004, 29(2): 172-175.
18、Wu Jian, Lei Moxian. New consideration on the relation of insulin resistance and type 2 diabetic. Medicine and Philosophy, 2004, 25(2): 4-5.
19、Mao Caipin, Gu Zhenlun. Experimental studies of effects of puerarin on hemorheology in diabetic rats. Chinese Journal of Hemorheology. 2003, 13(4): 322-324.
20、Greene DA. Acute and chronic complications of diabetes mellitus in older patients. American Journal of medicine. 1986, 80(5A): 39-53.
21、Chen JB, Xu JL, Jiang H, et al. The effects of puerarin on endothelin and blood platelet in patients suffering from coronary heart disease. Zhejiang J Tradit Chin Med (in Chinese). 1999, 34(5): 216-217.
22、Lu XR, Gao E, Xiu LZ, et al. Blocking effect of puerarin on β-adrenoceptor of isolated organs and whole animal. Acta Pharmacol Sin. 1986, 7(6): 537-539.
23、Zhang HY, Xu YB, Wang XJ, et al. Effects of puerarin on function of platelet and microalbuminuria in diabetes mellitus. Chin J New Drugs Clin Rem. 1999, 18(5): 291-293
24、Wang Jian, Chen Min, Cao Yi, et al. Effect of Kakonein and Gingko Flavone on Protein Nonenzymatic. Parmacology and Clinical Applications of Chinese Traditional Medicine. 2000, 16(1):13-14.
25、Zhang J T. Modern Methodology in Pharmacological Experiment. Beijin: Peking Medical College and Peking Union Medical College Publishing House, 1998: 981-982.
26、Li G W, Pan X R, Stephen L, et al. A new insulin-sensitivity index for the population-based study. Chin J Intern Med. 1993, 32(10): 656-660.
27、Moller DE, Flier JS. Insulin resistance mechanism, syndromes, and impli cations. N Eng J med, 1991, 325(13): 928-948.
28、Storlien LH, James DE, Burleigh KM, et al. Fat feeding causes widespread in vivo insulin resistance, decreased energy expenditure, and obesity in rats . AmJ Physiol, 1986, 251: E576-583.
29、Ding Shiying, Shen Zhufang, Chen Yueteng, et al. Using euglycemic clamp technique estimate two type of insulin-resistance animal model. Chinses Diabetes magazine, 2001, 9 (5): 286-289.
30、Li Chenzhong, Zhang Suhua, Shu Changda, et al. High fat meal induce insulin-resistance rats. Basic Medical Sciences and Clinics, 2000, 20(3): 93-95.
31、Chen Qi. Pharmacology of Trasitional Chinese Medicine Experiment Method, Dumb Millions Sanitation Publishing Company, 1993: 719.
32、Yin Jun, Chen MingDao, Zhou LiBing, et al. Effects of long-term high-fat diet on the occurrence of diabetes in rats. Journal of Chinese Physician, 2001, 6(1):47-49.
33、Chen Lin, Bai Jie. Influence of Fibrin, Chromium and Zinc on glucose tolerance of rats, Liaoning Journal of Utility Diabetic. 2001, 9(1):28-29.
34、Sakamoto S, Minami K, Niwa Y, et al. Effect of exercise training and food restriction on endothelium dependent Re2 laxation in the otsuka Long Evans Tokushima fatty rat, a model of spontaneous NIDDM. Diabetes, 1998, 47(1): 82.
35、Miles PDG, Higo K, Romeo OM, et al. Troglitazone prevents hyperglycemia induced but not glucosamine induced insulin resistance. Diabetes, 1998, 47(3): 395.
36、Du Jian, Zhao YuYan, Xie Hui, Zhao Jin Establishment and evaluation of animal model of insulin resistance with high fatty diet. J Chin Med Univ, 2002, 31(5):343-346.
37、Hevener AL, Reichart D, Olefsky J, et al. Thiazolidinedione treatment prevents free fatty acid-induced insulin resistance in male wister rats. Diabetes, 2001, 50: 2316-2322.
38、Mannucci E, Bardini G, Ognibene et al. Comparison between insulin sensitivity indexes in obese patients. Diabetes Care, 2000, 23(7): 1042-1043.
39、Bokemark L, Froden A, Attvall S, et al. The euglycemic hyperinsulinmic clamp examination: variability and reproducibility. Scand J Clin Lab Invest, 2000, 60(1):27-36.
40、Yang YongMei, Zhang NanYan. Effect of free fatty acids on diabetes fat toxicity. Chinese Journal of Misdiagnostics, 2003, 3(1):48.
41、Hui, Zhang Yah. High fatty feed induce insulin resistance animal model and estimate. Chinese Journal of Misdiagnostics, 2003, 3(10): 1531-1532.
42、Goetze S, Blaschke F, Stawowy P, et al. Biochem Biophys ResCommun, 2001, 287(3): 662-670.
43、Sheu WHH, lee WJ, Chang RL, et al. Clin Exp Hypertens, 2000, 22(6): 595-606.
44、Fruhbeck C, Gomez-Ambrosi J, Muruzabal FJ, et al. The adipocyte, a model for integration of endocrine and metabolic signaling in energy metabolism regulation. Am J Physiol Endocrinol Metab, 2001, 280: 827.
45、Cohen PG. Aromatase, adiposity, aging and disease. The hypogonadal-metabolic-atherogenic-disease and aging connection. Med Hypotheses, 2001, 56(6):702-708.
46、Jung M, et al. Infulence of varied PH cultured media on nitric oxide synthesis of hepatocytes. Chin J Dig, 2000, 20(1)26-29.
47、D Ambrosio SM, Gibson-D Ambrosio RE, Brady T, et al. Mechanisms of nitric oxide-induced cytotoxicity in Normal human hepatocytes. Environ Mol Mutagen, 2001, 37(1):46-54.
48、Liu GuoQing. Study headway on NO and diabetes syndrome. J Overseas Physic Incretion Fascicule, 1999, 19(6):257-259.
49、Li ChenZhong, Zhang SuHua, Su ChangDa, et al. Establishing and identifing insulin-resistance cell line. Acta Universitatis Scientiae Medicine ChongQing, 1998, 23:335-337.
50、Li ChangGui, Ning Guang, Chen JiaLun. Establishing and identifying insulin-resistant HepG2 cell line. Chinese Journal of Diabetes, 1999, 7(4):198-200.
51、Ying Jun, Hu RenMing, Chen MingDao, et al. Compare the effect of glucose disappeared on metformin and berberine on HepG2 cell. Chin J Endocrinol Metab, 2002, 18(6):488-489.
52、Hansen MB, Nielsen SE, Berg K. Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J Immunol Method, 1989, 119:203-210.
53、 Wang XiaoYun, Liao KunLing, Lu Wei, et al. The relationship between NO and IR of DM (Type Ⅱ). Guizhou Medical Journal, 2001, 25(1):18-19.
54、 Kuo WL, Gehm BD, Rosner MR, et al. Regulation of insulin degradation: expression of an evolutionarily conserved insulin degrading enzyme increase degradation by an intracellular pathway. Mol Endocrinol, 1991, 5:1467-1476.
55、 Authier F, Ponsner BF, Bergeron JJ. Insulin-degrading enzyme. Clin Invest Med, 1996, 19(2) :149-152.
56、 Michihiro Fukushima, Masuo Nakano, Yasuko Morii, et al. Hepatic LDL Receptor mRNA in Rats Is Increased by Dietary Mushroom (Agaricus bisporus) Fiber and Sugar Beet Fiber. Journal of Nutrition, 2000, 130:2151-2156.
57、 Caro J F, Amutrvda J M. Glucocorticoid induced insulin resistance of man. J Clin Invest, 1982, 69:866.
58、 Dgawa A. Roles of insulin resistance and β -cell dysfuntion in Dexamethasone-indncel diabetes. J Clin Invest, 1992, 90:497.
59、 Campbell K L, Latimer KS. Transient diabetes mellitus associated with prednisone therapy in dog. J Am Vet Med Assoc, 1984,185:299.
60、 Moller DE, Flier JS. Insulin resistance mechanism, syndromes, and implications. N Eng J med, 1991,325(13): 928-948.
61、 Kragen EW, Clark PW, Jenkins AB, et al. Development of muscie insulin resistance in high-fat-fed rats. Diabetes, 1991, 40:1397-1403.
62、 Storlien LH, Higgins JA , Thomas TC , et al. Diet composition and insulin action in animal models. British Journal of Nutrition, 2000, 83 (Suppll): S85-S90.
63、 Ye JM, Doyle PJ, Lglesias MA, et al. Peroxisome proliferator activated receptor (PPAR) alpha activation lowers muscle lipids and improves insulin sensitivity in high fat-fed rats: comparison with PPAR-gamma activation. Diabetes, 2001, 50(2):411-417.
64、Storlien LH, Oakes ND, Pan DA, et al. Syndrome of insulin resistan in the rats: inducement by diet and amelioration with Benfliorex. Diabetes, 1993, 42:457-462.
65、Yang GangYi, Peter Chung, Guenther Boden. Changes of glucose metabolism in insulin clamp studies in the non-anaesthesia rats. Basic Medical Sciences and Clinics. 2003, 23(3): 341-344.
66、Kraegen EW, James DE, Bennett SP, et al. In vivo insulinsensitivity in the rat determined by euglycemic clamp. Am J Physiol, 1983, 245 (Endocriol. Metab, 8): E1-7.
67、Yang GangYi, Li Ling, Peter Chung, Gangther Boden. Effects of phosphodiesterase inhibitor-induced insulin resistance on glucose and lipid metabolism in rats. Chinese Journal of Pathophysiology, 2003, 19(7): 875-880.
68、Lu HuiLing, Wang HongWei, Ling HanHua, Zhang MuXun. Relation of resistin with blood glucose, blood fattiness and insulin resistance on diabetes sufferer. Clin J Intern Med, 2004, 43(1):53-54.
69、Liang Zhen, Cheng Hua. Lipotoxicity and type 2 diabetes. J Overseas Physic Medicine Fascicule, 2004, 31(2):50-53.
70、Lupi R, Del Guerra S, Fierabracci V, et al. Lipotoxicity in human pancreatic islets and the protective effect of metformin. Diabetes, 2002, 51 Suppll:S134-137.
71、Mc Garry JD. Banting lecture 2001: dysregulation of fatty acid metabolism in the etiology of type 2 diabetes. Diabetes, 2002, 51: 7-18.
72、Cheung C, Clifton D, Steiner R. Proopiomelanocortin neurons are direct targets for leptin in the hypothalamus. Endocrinology, 1997, 138: 4489-4492.
73、Kieffer TJ, Heller RS, Leech CA, et al. Diabetes, 1997, 46: 1087-1093.
74、Stejskal D, Jedelsky L, Bartek J, et al. Vnitr Lek, 1998, 44:361-365.
75、Cao Xiaopei, Ming Wenyu, Cheng Hua, et al. Effects of leptin on glucose oxidation and glucokinase gene expression in cultured liver cells. Chin J Endocrinol Metab, 1999, 15(3): 131-133.
76、Kennedy A, Gettys TW, Watson P, et al. J Clin Endocrinol Metab, 1997, 82: 1293-1300.
77、L fgren P, Van Harmelen V, Reynisdottir S, et al. Diabetes, 2000, 49 (5): 688-692.
78、Fasshauer M, Klein J, Neumann S, et al. Biochem Biophys Res Commun, 2001, 288 (4): 1027-1031.
79、Nicaud V, Raoux S, Poirier O, et al. Atherosclerosis, 2002, 161 (2): 317-325.
80、Ronnemaa T, Pulkki K, Kaprio J, et al. J Clin Endocrinol Metab, 2000, 85 (1): 2728-2732.
81、Liu S, Spelleken M, R brig K, et al. Diabetes, 1998, 47(4):515-522.
82、Goetze S, Blaschke F, Stawowy P, et al. Biochem Biophys Res Commun, 2001, 287 (3): 662-670.
83、Sheu WH, Hlee WJ, Chang RL, et al. Clin Exp Hypertens, 2000, 22(6):595-606.
84、Nisoli E, Carruba MO, Tonello C, et al. Diabetes, 2000, 49(3):319-325.
85、Prins JB, Niesler CU, Winterford CM, et al. Diabetes, 1997, 46 (12): 1939-1944.
86、Rachael L, Fawcett, Angela S, et al. J Clin Endocrinol Metab, 2000, 85 (2): 530-535.
87、Zhu XiXing. β-cell dysfunction and insulin resistance. J Overseas Physic Incretion Fascicule, 2001, 21(4):169-176.
88、Weir CC, Laybutt DR, Kaneto H, et al. β-cell adaptation and decompensation during the progression of diabetes. Diabetes, 2001, 50(suppll): S154-159.
89、Shen Cuzhou. Insulin recept and insulin signal conduct access with insulin resistance. A cad J Sec Mil Med University, 2001, 22(11): 1062-1063.
90、Heck DE. NO, RSNO, ONOO-, NO-NOO, Nox-dynamic regulation of oxidant scavenging, nitric oxide stores, and cyclic GMP-independent cell signaling. Antioxid Redox Signal, 2001, 3(2): 249-260.
91、Molecular Clone Experiment Enchieidion. J Sambluker, et al. Jin DongYan, Li MengFeng translated. Beijing Science Publishing Company. 1996, 683-688.
92、Zeng G, Ouon MJ. Insulin-stimulated production of nitric oxide is inhibited by wortmannin. Direct measurement in vascular endothelial cells J Clin Invest, 1996; 98(4):894-898.
93、Zhang Hui, Chen XiangFang, Ye FuLin, Liu ZhiNian, et al. Effects of insulin on nitric oxide synthase (NOS) activity and the expression of inducible NOS mRNA in calf aortic endothelial cells. Chin J Endocrinol Metab, 2003, 19(6): 483-485.
94、Stuhlinger MC, Abbasi F, Chu JW, et al. Relationship between insulin resistance and an endogenous nitric oxide synthase inhibitor. JAMA, 2002, 287: 1420-1426.