氯沙坦改善2型糖尿病大鼠胰岛素抵抗的机制研究
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摘要
目的以高糖高脂饮食/STZ诱导的SD大鼠(Sprague-Dawley)为动物模型,研究在2型糖尿病大鼠中血管紧张素Ⅱ受体阻滞剂(ARB)改善胰岛素的代谢效应及对胰岛素引发的胰岛素受体底物1(IRS1)磷酸化、葡萄糖转运蛋白4(GLUT4)转位的影响,并研究其机制是否依赖于磷脂酰肌醇3激酶(PI3K)途径。方法将42只SD大鼠随机抽取20只作为正常对照组,给予普通饮食,其余为糖尿病模型组22只,给予高脂高糖饮食及链脲佐菌素(STZ),空腹血糖(FBG)≥7.8 mmol/L且伴有胰岛素抵抗者为造模成功,成模大鼠共20只,将正常对照组分为对照组(A组,n=10)和对照处理组(B组,n=10);将糖尿病模型组分为糖尿病组(C组,n=10)和糖尿病处理组(D组,n=10),B组及D组给予ARB类药物氯沙坦(losartan)处理6周,A组及C组给予等量的0.9%氯化钠溶液。干预6周后称体重,断尾取血测空腹血糖及胰岛素水平,计算胰岛素敏感指数,麻醉动物在注射胰岛素15 min后取下老鼠后腿肌肉,储存备用。用逆转录-聚合酶链式反应(RT-PCR)测GLUT4、IRS1、PI3K、Akt mRNA的表达。结果成功的制备了2型糖尿病大鼠模型,造模后,与A、B组比较,C、D组体重增加,空腹血糖、血浆胰岛素水平均升高(P<0.05),胰岛素敏感指数下降(P<0.05)。氯沙坦干预后,与C组相比,D组体重增加,空腹血糖、血浆胰岛素水平降低,胰岛素敏感指数升高(P<0.05)。RT-PCR结果显示:与A、B组比较,C、D组糖尿病大鼠GLUT4 mRNA表达降低(P<0.05),IRS1、PI3K、Akt mRNA表达差异无统计学意义(P>0.05)。氯沙坦干预后,与C组比较,D组糖尿病大鼠GLUT4 mRNA表达差异无统计学意义(P>0.05),IRS1、PI3K、AktmRNA表达差异无统计学意义(P>0.05)。A、B组比较,GLUT4、IRS1、PI3K、AktmRNA表达差异无统计学意义(P>0.05)。结论氯沙坦改善糖尿病大鼠IR,糖尿病处理组PtyrIRS1、GLUT4膜蛋白表达上升,Pser473-Akt的活性无差别,表明氯沙坦增加骨骼肌组织中GLUT4的转位有可能通过非PI3K途径。
Objective To investigate the effects of ARB in improving the metabolic effects of insulin and effects on phosphorylation of IRS1 induced by insulin and transposition of GLUT4,and to explore whether its action mechanism depends on the pathway of PI3 K. Methods Forty four SD rats were randomly divided into normal control group( n = 20) and model group( n = 22). The rats normal control group were fed with standard diet,however,the rats in model group were fed with high-fat and high-carbohydrate diet together with streptozotocin( STZ). The rats with fasting plasma glucose( FPG) ≥7. 8 mmol/L and insulin resistance were regarded as type 2 diabetic models. The rats in normal control group were randomly subdivided into control group A( n = 10) and treatment group B( n = 10),and 20 diabetic rats were randomly divided into NIDDM group C( n = 10) and NIDDM treatment group D( n = 10). The rats in group B and group D were treated with losartan for 6 weeks while the rats in group A and group C were treated with equivalent volume of 0. 9% sodium chloride solution. Six weeks later,the rats were weighted,blood samples were collected to record glucose and insulin levers for calculation of ISI( insulin sensitivity index). Then the rats were fasted overnight and anesthetized for fifteen minutes before insulin injection. skeletal muscle was quickly removed from hind legs and stored at 80℃. RT PCR was used to detect the expression levels of GLUT4,IRS1,PI3 K and AKT mRNA. Results As compared with those in group A and B,the body weight in group C and D was significantly increased,moreover,the levels of FPG,FINS were also significantly increased,however,ISI was decreased significantly( P < 0. 05). After intervention with losartan,as compared with that in group C,the body weight in group D was increased,but the levels of FPG and FINS were decreased,moreover ISI was significantly increased( P < 0. 05). As compared with those in group A and B,the expression levels of GLUT4 mRNA in group C and D were significantly decreased( P < 0. 05),however,there were no differences in the expression levels of IRS1,PI3 K and AKT( P > 0. 05). After intervention w ith losartan,th ere w ere n o d ifferen ce in th e expression levels of GLUT4 mRNA between group C and group D( P > 0. 05),moreover there were no significant differences in the expression levels of IRS1,PI3 K and AKT between the two groups( P > 0. 05). In addition there were no difference in the expression levels of GLUT4 mRNA,IRS1,PI-3 K and AKT between group A and group B( P >0. 05). Conclusion The losartan can improve IR of diabetic rats,increase the expression levels of Ptyr-IRS1 and GLUT4,and the action mechanism may be correlated with non PI3 kinase pathway.
Objective To investigate the effects of ARB in improving the metabolic effects of insulin and effects on phosphorylation of IRS1 induced by insulin and transposition of GLUT4,and to explore whether its action mechanism depends on the pathway of PI3 K. Methods Forty four SD rats were randomly divided into normal control group( n = 20) and model group( n = 22). The rats normal control group were fed with standard diet,however,the rats in model group were fed with high-fat and high-carbohydrate diet together with streptozotocin( STZ). The rats with fasting plasma glucose( FPG) ≥7. 8 mmol/L and insulin resistance were regarded as type 2 diabetic models. The rats in normal control group were randomly subdivided into control group A( n = 10) and treatment group B( n = 10),and 20 diabetic rats were randomly divided into NIDDM group C( n = 10) and NIDDM treatment group D( n = 10). The rats in group B and group D were treated with losartan for 6 weeks while the rats in group A and group C were treated with equivalent volume of 0. 9% sodium chloride solution. Six weeks later,the rats were weighted,blood samples were collected to record glucose and insulin levers for calculation of ISI( insulin sensitivity index). Then the rats were fasted overnight and anesthetized for fifteen minutes before insulin injection. skeletal muscle was quickly removed from hind legs and stored at 80℃. RT PCR was used to detect the expression levels of GLUT4,IRS1,PI3 K and AKT mRNA. Results As compared with those in group A and B,the body weight in group C and D was significantly increased,moreover,the levels of FPG,FINS were also significantly increased,however,ISI was decreased significantly( P < 0. 05). After intervention with losartan,as compared with that in group C,the body weight in group D was increased,but the levels of FPG and FINS were decreased,moreover ISI was significantly increased( P < 0. 05). As compared with those in group A and B,the expression levels of GLUT4 mRNA in group C and D were significantly decreased( P < 0. 05),however,there were no differences in the expression levels of IRS1,PI3 K and AKT( P > 0. 05). After intervention w ith losartan,th ere w ere n o d ifferen ce in th e expression levels of GLUT4 mRNA between group C and group D( P > 0. 05),moreover there were no significant differences in the expression levels of IRS1,PI3 K and AKT between the two groups( P > 0. 05). In addition there were no difference in the expression levels of GLUT4 mRNA,IRS1,PI-3 K and AKT between group A and group B( P >0. 05). Conclusion The losartan can improve IR of diabetic rats,increase the expression levels of Ptyr-IRS1 and GLUT4,and the action mechanism may be correlated with non PI3 kinase pathway.
引文
1娄志杰,韩向莉,邵岩.糖尿病肾病中医病因病机及中医药内、外治疗研究新进展.医学理论与实践,2017,30:808-810.
2 沈奇伟,张旭,姚琪远.糖尿病与周期基因研究进展.中华肥胖与代谢病电子杂志,2017,3:42-45.
3 毛黎静,安春红,张贝娜,等.新型糖尿病治疗药物的研究进展.中南药学,2017,15:89-91.
4 周方圆,杨宇峰,石岩.2型糖尿病胰岛素与胰岛素抵抗信号转导通路研究进展.辽宁中医药大学学报,2016,18:71-73.
5 任毅,左之才,万涛梅.抵抗素在胰岛素抵抗中的作用机制及其受体信号通路研究进展.生理学报,2016,68:65-74.
6 张泽生,李雨蒙,张梦娜,等.胰岛素抵抗细胞模型建立及药物评价的研究进展.中国食品添加剂,2016,7:198-203.
7 刁红杰,鲁燕.与胰岛素抵抗相关的基因多态性研究进展.实用糖尿病杂志,2016,12:3-5.
8 章鸿雁,黄琪,卜瑞芳.肾素血管紧张素系统与糖尿病性骨质疏松症的研究进展.中国骨质疏松杂志,2017,23:112-129.
9 Nickenig G1,R9ling J,Strehlow K,et al.Insulin induces upregulation of vascular AT1 receptor gene expression by posttranscriptional mechanisms.Circulation,1998,98:2453-2460.
10 Malhotra A1,Kang BP,Cheung S,et al.Angiotensin II promotes glucose-induced activation of cardiac protein kinase C isozymes and phosphorylation of troponin I.Diabetes,2001,50:1918-1926.
11 周亦秋,杨国君.RAS与胰岛素抵抗——代谢综合征新的治疗靶点.心血管病学进展,2006,27:753-755.
12 范秋灵,杨刚,刘晓丹,等.氯沙坦对自发性2型糖尿病KKAy小鼠肾小球蛋白表达谱的影响.中华肾脏病杂志,2012,28:476-483.
13 闫朝丽,王慧,孟兴凯,等.氯沙坦通过增加葡萄糖转运蛋白4膜转位改善2型糖尿病大鼠胰岛素抵抗.中华糖尿病杂志,2014,6:42-45.
14 徐丹,黄龙贤,周大燕,等.瑞格列奈分别联合氨氯地平、氯沙坦钾治疗T2DM合并原发性高血压的疗效和安全性比较.中国药房,2017,28:327-330.
15 唐纪兰,文新年,韦凌云,等.吡格列酮对非酒精性脂肪性肝病患者胰岛素抵抗及内皮功能的影响.中华实用诊断与治疗杂志,2015,29:303-307.
16 董秀丽,刘明岭,王晶,等.微量泵泵注胰岛素治疗糖尿病酮症酸中毒起始剂量分析.中华实用诊断与治疗杂志,2016,30:1238-1241.
17 胡俊锋,张先娇,郑达鑫,等.肥胖/糖尿病与肿瘤的相关性及其分子机制的研究进展.现代生物医学进展,2017,17:177-180.
18 曾理,丁群芳,许婷媛,等.DNA双链损伤修复机制在糖尿病致动脉粥样硬化中的作用研究.四川大学学报(医学版),2017,48:191-196.
19 张玲,牛建生.依那普利联合氯沙坦治疗早期2型糖尿病肾病合并高血压的疗效与安全性.中国老年学杂志,2013,33:2038-2040.
20 Derosa G,Querci F,Franzetti I,et al.Comparison of the effects of barnidipine plus losartan compared with telmisartan plus hydrochlorothiazide on several parameters of insulin sensitivity in patients with hypertension and type 2 diabetes mellitus.Hypertension research:Official journal of the Japanese Society of Hypertension,2015,38:690-694.
21 黄祺,朱圣炜,浦丹凤,等.初诊2型糖尿病患者胰岛素抵抗与肾素血管紧张素醛固酮系统的关系.实用医学杂志,2016,32:3299-3302.
22 杨强,何燕铭,王文健.肾素血管紧张素系统、胰岛素抵抗与盐敏感性高血压关系的研究进展.医学综述,2014,20:202-204.
2 沈奇伟,张旭,姚琪远.糖尿病与周期基因研究进展.中华肥胖与代谢病电子杂志,2017,3:42-45.
3 毛黎静,安春红,张贝娜,等.新型糖尿病治疗药物的研究进展.中南药学,2017,15:89-91.
4 周方圆,杨宇峰,石岩.2型糖尿病胰岛素与胰岛素抵抗信号转导通路研究进展.辽宁中医药大学学报,2016,18:71-73.
5 任毅,左之才,万涛梅.抵抗素在胰岛素抵抗中的作用机制及其受体信号通路研究进展.生理学报,2016,68:65-74.
6 张泽生,李雨蒙,张梦娜,等.胰岛素抵抗细胞模型建立及药物评价的研究进展.中国食品添加剂,2016,7:198-203.
7 刁红杰,鲁燕.与胰岛素抵抗相关的基因多态性研究进展.实用糖尿病杂志,2016,12:3-5.
8 章鸿雁,黄琪,卜瑞芳.肾素血管紧张素系统与糖尿病性骨质疏松症的研究进展.中国骨质疏松杂志,2017,23:112-129.
9 Nickenig G1,R9ling J,Strehlow K,et al.Insulin induces upregulation of vascular AT1 receptor gene expression by posttranscriptional mechanisms.Circulation,1998,98:2453-2460.
10 Malhotra A1,Kang BP,Cheung S,et al.Angiotensin II promotes glucose-induced activation of cardiac protein kinase C isozymes and phosphorylation of troponin I.Diabetes,2001,50:1918-1926.
11 周亦秋,杨国君.RAS与胰岛素抵抗——代谢综合征新的治疗靶点.心血管病学进展,2006,27:753-755.
12 范秋灵,杨刚,刘晓丹,等.氯沙坦对自发性2型糖尿病KKAy小鼠肾小球蛋白表达谱的影响.中华肾脏病杂志,2012,28:476-483.
13 闫朝丽,王慧,孟兴凯,等.氯沙坦通过增加葡萄糖转运蛋白4膜转位改善2型糖尿病大鼠胰岛素抵抗.中华糖尿病杂志,2014,6:42-45.
14 徐丹,黄龙贤,周大燕,等.瑞格列奈分别联合氨氯地平、氯沙坦钾治疗T2DM合并原发性高血压的疗效和安全性比较.中国药房,2017,28:327-330.
15 唐纪兰,文新年,韦凌云,等.吡格列酮对非酒精性脂肪性肝病患者胰岛素抵抗及内皮功能的影响.中华实用诊断与治疗杂志,2015,29:303-307.
16 董秀丽,刘明岭,王晶,等.微量泵泵注胰岛素治疗糖尿病酮症酸中毒起始剂量分析.中华实用诊断与治疗杂志,2016,30:1238-1241.
17 胡俊锋,张先娇,郑达鑫,等.肥胖/糖尿病与肿瘤的相关性及其分子机制的研究进展.现代生物医学进展,2017,17:177-180.
18 曾理,丁群芳,许婷媛,等.DNA双链损伤修复机制在糖尿病致动脉粥样硬化中的作用研究.四川大学学报(医学版),2017,48:191-196.
19 张玲,牛建生.依那普利联合氯沙坦治疗早期2型糖尿病肾病合并高血压的疗效与安全性.中国老年学杂志,2013,33:2038-2040.
20 Derosa G,Querci F,Franzetti I,et al.Comparison of the effects of barnidipine plus losartan compared with telmisartan plus hydrochlorothiazide on several parameters of insulin sensitivity in patients with hypertension and type 2 diabetes mellitus.Hypertension research:Official journal of the Japanese Society of Hypertension,2015,38:690-694.
21 黄祺,朱圣炜,浦丹凤,等.初诊2型糖尿病患者胰岛素抵抗与肾素血管紧张素醛固酮系统的关系.实用医学杂志,2016,32:3299-3302.
22 杨强,何燕铭,王文健.肾素血管紧张素系统、胰岛素抵抗与盐敏感性高血压关系的研究进展.医学综述,2014,20:202-204.