摘要
研究目的
本研究是导师课题的一部分,主要观察针刺对胰岛素抵抗模型大鼠骨骼肌IRS-1、IRS-2、GLUT4的mRNA和蛋白表达的影响,从分子水平探讨针刺调整胰岛素抵抗的机理。
研究方法
将24只雄性SD大鼠(180-220g)按体重随机分为3组,每组8只,分别为空白组、模型组和针刺组。空白组予普通饲料喂养,其余各组予高糖高脂高盐饲料喂养。从第6周开始,每隔1周对空白组和模型组大鼠从眼眶静脉窦采血1次,检测空腹血糖(FPG)、血浆胰岛素(FINS),并计算胰岛素敏感性指数(ISI),当模型组大鼠ISI较空白组显著降低(P<0.01)为造模成功。第12周造模成功后,空白组继续予普通饲料喂养3周,模型组继续予高脂高糖高盐饲料喂养3周,针刺组继续予高脂高糖高盐饲料喂养3周,同时针刺组给予电针治疗日1次,取内关、足三里、三阴交和肾俞穴。治疗结束后各组大鼠禁食12小时过夜,次晨行眼眶静脉窦采血,检测FPG、FINS、C肽等指标,FPG测定采用葡萄糖氧化酶法,FINS、C肽测定采用ELISA法,ISI用1/(CFPG X JFINS)公式计算;胰岛素灌注后将大鼠处死,取股四头肌,用RT-PCR、Western blotting等检测手段,观察针刺前后胰岛素抵抗模型大鼠骨骼肌IRS-1、IRS-2、GLUT4的mRNA和蛋白表达情况。
结果
予高脂高糖高盐饲料喂养12周后,模型组大鼠和针刺组大鼠的FPG、FINS与空白组相比均显著升高(P<0.01), ISI显著下降(P<0.01),提示造模成功。经过3周针刺治疗后,针刺组的FPG、FINS、C肽与模型组比较显著降低(P<0.01, P<0.05), ISI与模型组相比显著升高(P<0.01)。
模型组大鼠骨骼肌IRS-1 mRNA表达较空白组显著降低(P<0.01);针刺组大鼠骨骼肌IRS-1 mRNA较模型组显著升高(P<0.01)。
模型组大鼠骨骼肌IRS-2 mRNA表达较空白组显著降低(P<0.01), IRS-2蛋白表达与空白组相比无明显变化(P>0.05);针刺组大鼠骨骼肌IRS-2 mRNA和IRS-2蛋白表达均较模型组显著升高(P<0.01)。
模型组大鼠骨骼肌GLUT4 mRNA和GLUT4蛋白表达均较空白组显著降低(P<0.01);针刺组大鼠骨骼肌GLUT4 mRNA和GLUT4蛋白表达均较模型组显著升高(P<0.01)。
结论
研究结果提示,针刺改善胰岛素抵抗主要是通过调节PI3K通路的信号转导来实现,其机理与增加IRS-1、IRS-2、GLUT4 mRNA表达和GLUT4蛋白表达有关。
Objective
Our aim was to observe the effects of acupuncture on IRS-1, IRS-2 and GLUT4 of insulin resistance model rats' femoris muscle, so as to further explore its underlying mechanism.
Methods
Twenty four male SD rats were divided randomly into three groups:the blank group, the model group and the acupuncture group. We induced insulin resistance model through the high glucose, high salt and high fat diet. From the sixth week, we tested FPG and FINS in the rat's blood every week, then figured out the ISI. When the ISI of the model group and the blank group was statistically significant (P< 0.01), the model was success. Then in the next three weeks, the rats of the blank group were fed by normal diet, while the rats of the model group and the acupuncture group were fed by the high glucose, high salt and high fat diet. The rats of the acupuncture group were treated by electroacupuncture at the same time, twenty minutes per day. We chose four points:Neiguan, Zusanli, Sanyinjiao and Shenshu.
After three weeks of treatment, all rats were drew blood, preparing for testing FPG, FINS and C-P. Then they were perfused by insulin before being killed. Their quadriceps femoris muscle was cut off. Finally IRS-1, IRS-2 and GLUT4 were tested by methods such as RT-PCR, western blotting and so on.
Results
Insulin resistance rat model was built successfully. FPG and FINS of the model group rose evidently(P<0.01), ISI of the model group declined remarkably(P<0.01). After treatment, FPG, FINS, ISI and C-P of the acupuncture group improved evidently(P<0.01, P<0.05). IRS-1 mRNA of the model group declined remarkably (P<0.01). IRS-1 mRNA of the acupuncture group rose evidently (P<0.01).
IRS-2 mRNA of the model group declined remarkably (P<0.01). IRS-2 mRNA and IRS-2 protein expression of the acupuncture group rose evidently (P<0.01). GLUT4 mRNA and GLUT4 protein expression of the model group declined remarkably (P<0.01). GLUT4 mRNA and GLUT4 protein expression of the acupuncture group rose evidently (P<0.01).
Gonelusion
In this study, we found acupuncture's regulating action to PI3K route. We presume acupuncture treat insulin resistance model rats through increasing IRS-1 mRNA, IRS-2 mRNA and protein expression, GLUT4 mRNA and protein expression. The action mode and pathway of these protein were not clear yet, which need us to study further.
引文
[1]Reaven GM. Role of insulin in human disease[J]. Diabetes,1988,37(12):1595-1607.
[2]Reaven GM. Role of insulin resistance in human disease (syndrome X):an expanded definition[J]. Annu Rev Med,1993,44:121-131.
[3]Stem MP. Diabetes and cardiovascular disease, The "common soil" hypothesis[J]. Diabetes, 1995,4:369-374.
[4]Cusi K, Maezono K, Oaman A et al. Insulin resistance differentially affects the pi3-kinase-and MAP kinase-me-diated signaling in human muscle[J]. J Clin Invest,2000,105: 311-320.
[5]Anna D, Anita O, Fredrik H. Attenuation of Insulin-stimulated Insulin Receptor Substrate-1 Serine 307 Phosphorylation in Insulin Resistance of Type 2 Diabetes[J]. J Biol Chem,2005, 280:34389-34392.
[6]Jennifer J, Thierry G, Mireille C. Interleukin-1-Insulin Resistance in Adipocytes through Down-Regulation of Insulin Receptor Substrete-1 Expression[J]. Endocrinology,2007,148: 241-251.
[7]Avia H, Yan-Fang L, Erez I. Common Inhibitory Serine Sites Phosphorylated by IRS-1 Kinases, triggered by Insulin and Inducers of Insulin Resistance [J]. J Biol Chem,2007,280: 18018-18027.
[8]Withers D J, white M. The insulin signaling systema comm on link in the pathogenesis of type 2 diabetes. Endocrinology.2000,141:1917-1921.
[9]Kadowaki T. Insights into insulin resistance and type 2 diabetes from knockout mouse models[J]. J Clin Invest,2000,106:459-465.
[10]Gang Y, Juanjuan D, Tao W. Tissue Kallikrein Reverses Insulin Resistance and Attenuates Nephropathy in Diabetic Rats by Activation of Pathosphatidylinositol 3-Kinase/Protein Kinase B and Adenosine5'-Monophosphate-Activated Protein Kinase Signaling Pathways[J]. Endocninology,2007,148:2016-2026.
[11]Karen T, Wendy A K, Jianan L. Analysis of Genetic Variation in Akt2/PKB-pin Severe Insulin Resistance, Lipodystrophy, Type 2 Diabetes, and Related Metabolic Phenotypes[J]. Diabetes,2007,56:714-719.
[12]Vollenweider P, Menard B, Nicod P. Insulin resistance, defective insulin receptor substrate 2-associated phosphati-dylinositol-3'kinase activation, and impaired atypical protein kinase C (zeta/lambda) activation in myotubes from obese patients with impaired glucose tolerance[J]. Diabetes,2002,51:1052-1059.
[13]Zhanguo G, Zhong W, Xiaoying Z. Inactivation of PCK leads to increased susceptibility to obesity and dietary insulin resistance in mice[J]. Am J Physiol Endocrinol,2007,292: E84-E91.
[14]高磊,刘绛光.对葡萄糖转运蛋白的讨论[J].国外医学临床生物化学与检验学分册,1999,20(6):274-275.
[15]Maureen J C, Naira G, J SkyeL. Use of GLUT4 null mice to study skeletal muscle glucose uptake[J]. Clinical and Experimental Pharmacology and Physiology,2005,32:308-313.
[16]Hakam A, Adrian C, Jan F. Two phases of palmitate-induced insulin resistance in skeletal muscle:impaired GLUT4 translocation is followed by a reduced GLUT4 intrinsic activity [J]. Am J Physiol Endocrinol Metab,2007,293:E783-E793.
[17]郑仲华,曾庆明,李玲.从脾虚痰瘀论治2型糖尿病胰岛素抵抗[J].中医研究,2007,20(11):15-16.
[18]戴小良,王行宽.浅谈中医胰与糖尿病的关系[J].辽宁中医杂志,2002,29(1):27.
[19]第五永长,许建秦.胰岛素抵抗中医病机探讨[J].四川中医,2008,26(1):8-9.
[20]王利明.从痰湿论治胰岛素抵抗[J].现代中西医结合杂志,2003,12(7):783.
[21]李道本,王智明.从肝论治防治胰岛素抵抗治疗2型糖尿病的理论探讨[J].中国中西医结合消化杂志,2001,9(3):153-154.
[22]吴深涛.对胰岛素抵抗宜用益肾化瘀、疏利少阳法[J].中医杂志,2001,42(6):332-333.
[23]陈维铭,王馨然.半夏白术天麻汤对单纯性肥胖患者胰岛素抵抗的影响[J].现代中西医结合杂志,2004,13(2):351.
[24]陈彬,刘建,向楠.化痰降浊汤对糖调节受损的临床研究[J].湖北中医杂志,2008,30(9):11-13.
[25]尹义辉,王经武.降浊消糖饮治疗胰岛素抵抗疗效观察[J].山东中医杂志,2005,24(11):653.
[26]武明东,高天舒,郑曙琴等.化痰解瘀汤联合胰岛素治疗2型糖尿病疗效观察[J].中医药学刊,2005,23(4):729-730.
[27]赵纪剐.复方丹参注射液治疗2型糖尿病患者胰岛素抵抗的临床研究[J].河北中医,2007,29(6):549.
[28]薛青,谢丹红,卫苓等.清肝泻心汤对2型糖尿病胰岛素抵抗相关因素影响的研究[J].第三军医大学学报,2007,29(7):613.
[29]陆灏,朱宇清,唐静芬,等.三黄煎对2型糖尿病大鼠胰岛素分泌等功能的影响[J].辽宁中医杂志,2000,27(6):281.
[30]何月光,唐眷娥,罗炳辉等.黄芪及其注射液对冠心病患者胰岛素、血糖和血脂影 响的研究[J].现代诊断与治疗,2005,16(4):221-223.
[31]张帆,洪秀珍,刘东风.葛根素对不稳定心绞痛患者胰岛素抵抗血脂及QT离散度的影响[J].现代中医药,2007,27(5):3-4.
[32]孙文才,周和平,徐清朴等.葛根素对粥样硬化性肾动脉狭窄患者胰岛素抵抗的影响[J].中国中医急症,2007,16(12):1471-1472.
[33]于健.葛根素治疗2型糖尿病患者胰岛素抵抗的临床研究[J].山东中医杂志,2001,20(12):727·729.
[34]江焱.黄连素对代谢综合征胰岛素敏感性的影响[J].浙江中医药大学学报,2009,33(3):390.
[35]陈洪源,明智强,谢佳乐等.小檗碱降血糖机制的研究进展[J].食品与药品,2008,10(3):69—71.
[36]于志清,简锦溢,林碧霞等.番石榴叶治疗糖尿病166例临床观察阴广西医科大学学报,1980,(01).
[37]Yong Wu, Jing Ping, Ou Yang et al.Hypoglycemic effect of Astragalus polysaccharide and its effect on PTP-1B[J]. Acta Pharmacologica Sinica,2005,26(3):345-352.
[38]刘敏,欧阳静萍,吴珂等.黄芪多糖对KKAy小鼠骨骼肌蛋白激酶B丝氨酸磷酸化的影响[J].武汉大学学报(医学版),2006,27(2):135-139.
[39]孙卫,郑学芝,崔荣军等.葛根素对2型糖尿病大鼠胰岛素抵抗及脂肪分化相关蛋白基因表达的影响[J].医药导报,2008,27(10):1159-1161.
[40]陈广,陆付耳,王增四等.小檗碱改善2型糖尿病大鼠胰岛素抵抗与PI3K、GLUT4蛋白相关性的研究[J].中国药理学通报,2008,24(8):1007-1010.
[41]李春梅,高永林,李敏等.知母皂苷对小鼠血糖的影响[J].中药药理与临床,2005,21(4):22-23
[42]刘永玉,毛良,莫启等.大黄对非胰岛素依赖型糖尿病大鼠胰岛素受体的作用[J].中华医学杂志,1991,71(6):350-351.
[43]戴玲,赵帜平,沈业寿等.丹皮多糖对葡萄糖苷酶作用的影响[J].生物学杂志,2006,23(2):23-24
[44]R Zhao, Q Li, B Xiao. Effect of Lycium barbarum polysaccharide the improvement of insulin resistance in NIDDM rats[J]. Yakugaku Zasshi,2005,125(12):981-983.
[45]胡国强,杨保华,张忠泉.山药多糖对大鼠血糖及胰岛素释放的影响[J].山东中医药杂志,2004,23(4):230-231.
[46]范文东,章浩军,陈益升.益气化浊除湿法治疗糖尿病胰岛素抵抗30例[J].江西中医药,2009,40(314):40-41.
[47]景常林.益胰降糖合剂治疗2型糖尿病胰岛素抵抗患者40例临床观察[J].世界中医药,2009,4(6):307-309.
[48]唐东晖,李俐,冯森坚.补阳还五汤加味治疗糖耐量减低患者IR的临床观察[J].黑龙江中医药,2006,(3):15-17.
[49]朱章志,熊曼琪,林安钟.三黄降糖方对2型糖尿病患者外周胰岛素抵抗的影响[J].中国中西医结合杂志,1997,17(10):590-593.
[50]高允珊.活血降糖方治疗2型糖尿病及胰岛素抵抗50例疗效观察[J].新中医,2004,36(10):37-38.
[51]陈国华,沈守祥,潘光辉,等.黄芪育阴煎干预2型糖尿病患者胰岛素抵抗及相关因素的临床研究[J].潍坊医学院学报,2006,23(3):13
[52]梁兴伦,韩明向,徐兆芳.四安胶囊治疗胰岛素抵抗综合征的临床研究[J].安徽中医学院学报,1997,16(1):16-20
[53]钱俊文,柴可夫.加减抵当汤防治大鼠胰岛素抵抗的疗效和机理研究[J].中国中医药科技,2007,14(6):396-397.
[54]丁来标,陆付耳,叶爱丽等.黄连解毒汤对胰岛素抵抗大鼠瘦素和抵抗素的影响[J].中国中西医结合杂志,2006,26(3):232-235
[55]李运伦.清热解毒法对SHR胰岛素抵抗的实验研究[J].中国中医急症,2007,16(11):1380-1381.
[56]姜淼,王世东,黄允瑜等.黄连人参对治疗2型糖尿病胰岛素抵抗机制的实验研究[J].新中医,2006,38(5):89-91
[57]王开成,方朝晖.丹蛭降糖胶囊对胰岛素抵抗大鼠脂联素的影响[J].江西中医学院学报,2006,8(12):61-63.
[58]张益钧,沈利水,戴盛锋等.三黄汤与普济消毒饮干预2型糖尿病小鼠胰岛素信号错误转导的机理研究[J].中医药学报,2009,37(2):25-28.
[59]夏卫军,金妙文.三地降糖片对实验性NIDDM大鼠的影响[J].江苏中医.2001,22(5):41-43
[60]张智龙,薛莉,吉学群等.针刺对2型糖尿病胰岛素抵抗影响的临床研究[J].中国针灸,2002,22(11):723-725.
[61]袁爱红,刘志诚,魏群利等.针灸治疗肥胖2型糖尿病35例[J].安徽中医学院学报,2009,28(1):28-30.
[62]曲齐生,杨善军.针刺夹脊穴对2型糖尿病患者胰岛素抵抗疗效观察[J].中医药信息,2007,24(1):50-51.
[63]刘志诚,孙凤岷,赵东红等.针刺对单纯性肥胖瘦素和胰岛素抵抗的影响研究[J].中医药学刊,2003,21(1):40-43.
[64]闫润虎,白洁,杨文波等.穴位埋线结合耳压疗法治疗单纯性肥胖病[J].中国临床康复,2006,10(11):8-10.
[65]赵东杰,范群丽.针刺对高血压病胰岛素抵抗的影响[J].中国针灸,2003,23(3): 165-167.
[66]郭玉红.针刺治疗高血压病胰岛素抵抗的临床观察[J].中医药学报,2007,35(6):51-53.
[67]郭玉红,滕秀英,张春芳等.针刺对原发性高血压患者胰岛素抵抗及血清TNF-a水平的影响[J].中医药学报,2009,37(5):76-78.
[68]刘淑杰,陆金平.针刺对缺血性脑血管病胰岛素抵抗的影响[J].辽宁中医学院学报,2005,7(6):562-563.
[69]唐胜修.头穴对中风后遗症患者胰岛素抵抗的调整作用研究[[J].安徽中医临床杂志,2002,14(3):164-165.
[70]盛鹏杰,吴娈灵,陈小张等.针灸治疗多囊卵巢综合征临床观察[J].湖北中医杂志,2010,32(2):65-66.
[71]易玮,许能贵,靳瑞.胰岛素抵抗机理及针刺干预的作用靶点[J].针刺研究,2002,27(1):73-75.
[72]易玮,许能贵,靳瑞.针刺对实验性大鼠胰岛素抵抗的逆转作用[J].广州中医药大学学报,2003,20(3):214-215.
[73]易玮,孙健,许能贵等.针刺对胰岛素抵抗模型大鼠肝脏和胰腺的形态学影响[J].新中医,2007,39(1):104-106.
[74]易玮,许能贵,孙健等.针刺对胰岛素抵抗模型大鼠血清胰岛素抗体和肿瘤坏死因子α的影响[J].中国针灸,2007,27(7):525-527.
[75]李嘉.针灸治疗非胰岛素依赖性糖尿病肥胖型的临床观察及作用机理研究.南京:南京中医药大学出版社,2000,南京中医药大学博士论文:17-22
[76]刘志诚,孙凤岷.魏群利等.针刺对非胰岛素依赖性糖尿病大鼠脂肪细胞葡萄糖摄取的影响[J].世界今日医学杂志,2001,2(8):667-669
[77]徐放明.针灸治疗非胰岛素依赖性糖尿病临床与机理研究.南京:南京中医药大学出版社,2001,南京中医药大学博士论文:18-24
[78]许能贵,梁兴伦.针刺对自发性高血压大鼠血压及胰岛索抵抗的效应研究[J].中国针灸,1997,(8):493.
[79]刘志诚,项晓人,袁锦虹等.针刺对NIDDM大鼠作用的形态学观察[J].现代中西医结合杂志,2002,11(17):1650-1652.
[80]曾志勇.针刺对糖尿病家兔血糖及胰高血糖素的影响[J].四川中医,2000,18(11):8-9.
[81]孙健,贾真,谢长才等.针刺对胰岛素抵抗模型大鼠肝脏PPARγ mRNA和蛋白的影响[J].辽宁中医杂志,2009,36(7):1195-1197.
[82]孙健,谢长才,贾真等.针刺对胰岛素抵抗模型大鼠游离脂肪酸含量的影响[J].山东中医药大学学报,2009,33(3):246-247.
[83]Kadowaki T. Insights into insulin resistance and type 2 diabetes from knock out mouse models[J]. J Clin Invest,2000,106(4):459-465.
[84]Kido Y, Burks D J, Withws D et al. Tissue specific insulin resistance in mice with mutations in the insulin receptor, IRS1, and IRS2[J]. J Clin Invest,2000,105(2):199-205.
[85]Penicaud L, Ferre P, Terretaz J et al. Development of obesity in Zucker rats:Early insulin resistance in muscles but normal sensitivity in white adipose rats[J]. Diabetes,1987,36(5): 626-631.
[86]Linda M, MurakamiT, Ishida K, et al. Phenotype-linked amino acid alteration in leptin receptor cDNA from Zucker fatty (fa/fa) rat[J]. Biochem Biophys Res Commun,1996,222: 19-26.
[87]Zhang Y, Proenca R, Maffei M, et al. Position cloning of the mouse obese gene and its human homologue[J]. Nature,1994,372:425-432.
[88]施新猷绾,医用实验动物学.西安:陕西科学技术出版社.1989:66-116.
[89]Kawano K, Hirashima T, Mori S, et al. Spontaneous Long-term hyperglycemic rat with diabetic complications-Ostuka Long-Evans Tokushima Fatty (OLETF) strain. Diabetes,1992, 41(11):1422.
[90]Ishida K et al. Which is the primary etiologic event in Ostuka Long Evans Tokushima Fatty rats, a model of spontaneous non-insulin-dependent diabetes mellitus, insulin resistance or impaired insulin secretion? [J] Metabolism,1995,44:940.
[91]Storlien L H, James D E, Burleigh K M et al. Fat feeding causes widespread in vivo insulin resistance, decreased energy expenditure, and obesity in rats[J]. Am J Physiol,1986, 251:576-583.
[92]邬云红,李秀钧,李宏亮等.高脂饮食肥胖大鼠胰岛细胞胰岛素抵抗机理的探讨[J].中华医学杂志,2005,85(27):1907-1910.
[93]Gianluca SB, Cinzia C, Stefania S, et al. A Model of Insulin and Nonalcoholic Steatohepatitis in Rats-Role of Peroxisome Proliferator Activated Receptorn and n3 Polyunsaturated Fatty AcidTreatment on LiverInjury[J]. Am J Pathol,2006,169(3):846-860
[94]卜石,杨文英,王昕等.长期高脂饲养对大鼠葡萄糖刺激的胰岛素分泌的影响[J].中华内分泌代谢杂志,2003,19(2):25-28.
[95]程海波,司晓晨,尚文斌等.高脂饲料和高果糖餐分别诱导胰岛素抵抗大鼠模型的胰岛素敏感性[J].中国临床康复,2006,10(7):121-123.
[96]张闽,杨晔,刘敏等.果糖诱导的高血压伴胰岛素抵抗大鼠血清脂联素肿瘤坏死因子-α度的变化[J].心肺血管病杂志,2007.26(1):48-50.
[97]吴海娅,魏丽,王深等.高脂或高糖饮食诱导胰岛素抵抗大鼠的附睾脂肪组织视黄醇结合蛋表达[J]. Shanghai Med J,2007,6(30):446-448.
[98]程海波,司晓晨,尚文斌等.高果糖餐诱导实验性胰岛素抵抗综合征大鼠模型[J].南京中医药大学学报,2006,22(1):31-34.
[99]潘玲,刘继林,王建等.实验性胰岛素抵抗综合征大鼠模型[J].华西医学,2000,15(4):421-422.
[100]洪丽莉,许冠荪,申国明等.SD大鼠2型糖尿病模型的建立[J].实验动物科学与管理,2005,22(4):1-3.
[101]沈亚非,徐焱成.高糖高脂膳食和链脲佐菌素诱导2型糖尿病模型的建立[J].实用诊断与治疗杂志,2006,20(9):649-651.
[102]孙静,宋光明,谢文利等.小剂量地塞米松诱发的胰岛素抵抗大鼠模型[J].武警医学院学报,2008,17(1):25-28.
[103]丁世英,申竹芳,谢明智等.MSG肥胖大鼠胰岛素抵抗特征的初步研究[J].中国药理学通报,2001,17(2):181-185.
[104]罗谋伦,郭欲晓,胡映辉等.罗格列酮的胰岛素增敏作用和胰岛素抵抗改善作用[J].中国药理学通报,2000,16(4):425-428.
[105]Saini KS, Thompson C, Winterford CM et al. Streptozotocin at low doses induces apoptosis and at high doses causes necrosis in a murine pancreatic beta cell line[J]. J Biochem Mol Biol lnt,1996,39(6):1229-1236.
[106]方朝晖,鲍陶陶,王开成等.丹蛭降糖胶囊对胰岛素抵抗大鼠PPAR-γmRNA表达的影响[J].中国实验方剂学杂志,2006,12(4):36-39.
[107]De Fronzo R A, Tobin J D, Anadres R. Glucose clamp technique:a method for quantifying insulin excretion and resistance[J]. Am J Physiol,1979,237(3):E214-223.
[108]Caro J F. Clinical review 26. Insulin resistance in obese and nonobese man[J]. J Clin Endocrinol Metab,1991,73:691-695.
[109]李光伟.胰岛素敏感性评估及其在临床研究中的应用[J].中华内分泌代杂志,2000,16(3):198-200.
[110]林文注,王佩.实验针灸学[M].上海:上海科学技术出版社.1994:286.[111]王国洪,许瑞吉,张中书等.血清C肽浓度测定临床意义探讨[J].放射免疫学杂志,2006,19(1):15-17.
[112]葛军,俞春芳,孙卫东等.不同空腹血糖状态人群的C肽值意义[J].上海医药,2005,28(6):472-474.
[113]Yaspelkis III B. B., Singh M. K., Trevino B. et al. Resistance training increases glucose uptake and transport in rat skeletal muscle[J]. Acta Physiol Scand 2002,175,315-323.
[114]Katsutaro Mofino, Kitt Falk Petersen, Sylvied Dufour, et al. Reduced m itochondrial density and increasedIRS-1 serine phosphorylation in muscle of insulin-resistant offspring oftype 2 diabetic parents [J]. Journal ofClinical Investigation,2005,115(12):3587-3595
[115]ANNA DANIELS SON, ANITA 0 ST, Erika Lystedt, et al.lnsulinresistance in human adipocytes occursdownstream of IRS-1 aftersurgical cell isolation but at thelevel of phosphorylation of IRS-1 in 1 type2 dia-betes[J]. THEFEBS Journal,2005,272:141-151
[116]HRIBAL ML, FEDERICI M, PORZIO 0, et al. The Gly Arg972 aminoacid polymorphism in insulin receptor substrate-1 afects glucosemetabolism in skeletal muscle cells [J]. J Clin Endocrinol Metab,2000:2004-2013
[117]PORZIOO, FEDERIC1 M, HRIBAL ML, et al. The G1y972Arg aminoacid polymorphism in IRS-1 impairs insulin secretion in pancreaticbeta cells [J]. J Clin Invest, 1999,104:357-364
[118]JELLEMA A, ZEEGERS MP, FESKENS EJ, et al. Gly972Argvariant in the insulin receptor substrate-1 gene and association withtype 2 diabetes:a metaanalysis of 27 studies [J]. Diabetologia,2003,46:990-995
[119]ELEFLHER1A ZEGGINI, JAMES PARKINSON, STEPHANIE HALFORD, et al. M cCarthy Association Studies of Insulin Receptor Substrate 1 Gene (IRS- 1)Variants in Type 2 Diabetes Samples Enriched for Family His-tory and Early Age of Onset[J]. Diabetes,2004, 53:33 19-3322
[120]WITHERS DJ, BURKS DJ, TOWERY HH, et al. Irs-2 coordinates Igf-1 receptor-mediated beta-cell development and peripheral insulin signaling[J]. Nat Gene,1999, 23:32-40
[121]MATTHEW J. Brady IRS-2 takes center stage in the development of type 2 diabetes[J]. Clin Invest,2004,114(7):886-888
[122]WITHERS D J, GUTIERREZ J S, TOWERY H, et al. Disruption of IRS-2 causes type 2 diabetes in mice [J]. Nature,1998,391:900-904
[123]WITHERS D J, BUNKS D J, TOWERY H, et al IRS-2 coordinate's lgf-1 receptor-mediated B cell development and peripheral insulin signal-ing[J]. Nal Genet,1999, 23:32-40
[124]BODEN C, SHUMAN G1. Fee fatty acid and type 2 diabetes defining their role in the development of insulin resistance and beta-cell dys-function[J]. Eur J Clin Invest.2002,32(3): 14-23
[125]VAZAROVA B, Weyer C, Hanson K, et al. Circulating interleukin-6 in relation to adiposity, insulin action, and insulin secretion [J]. Obs Res,2001,9(7):414-417
[126]田刚,周翔,刘巨永,等.2型糖尿病大鼠模型GLUT4mRNA表达的研究[J].天津医药,2005,23(8):511-513.
[127]ZHANG Liang(张梁),LI Mei(李梅),YI Wei(易玮),XU Neng—gui(许能贵),SUNJian(孙健). Effect on the Akt2 in Skeletal M uscle of Rats withInsulin Resistance Treated by Acupuncture. J. Acupunct. Tuina. Sci.2010,8(5):277-281
[128]张梁,李梅,易玮,许能贵,孙健.针刺对胰岛素抵抗模型大鼠骨骼肌PI3K p85的影响.新中医,2010,42(10):112-114