血管紧张素1-7在糖尿病及糖尿病肾病中与内质网应激之间的关系

发布时间：2018-04-01 12:26

本文选题：Ang1-7　切入点：糖尿病　出处：《天津医科大学》2017年硕士论文

【摘要】：目的:1.研究不同糖代谢状态人群的血管紧张素1-7(Ang 1-7)、血管紧张素转换酶(ACE)2水平,探讨其在糖尿病及其并发症中的作用。2.制备糖尿病肾脏模型,观察糖尿病大鼠肾脏病变和评价Ang 1-7对糖尿病大鼠肾脏内质网应激和凋亡的影响。方法:选取2014年12月于天津市海滨人民医院95名体检人员为研究对象,所有受试者均进行口服葡萄糖耐量试验,参照《中国2型糖尿病防治指南2010》2型糖尿病诊断标准:空腹血糖≥7.0mmol/L,餐后血糖≥11.1mmol/L。血糖异常患者包括空腹血糖受损(空腹血糖介于6.1~7.0 mmol/L)和餐后血糖调节受损(餐后2 h血糖介于7.8~11.1 mmol/L)的患者。根据血糖水平分为正常组(n=31)、糖耐量异常组(n=32)和糖尿病组(n=32)。检测其体重、腰围、血压、肝、肾功能、血脂水平、Ang 1-7、血管紧张素转换酶(ACE)2等指标,评价三组患者Ang 1-7水平及与血糖的关系,进一步应用多因素非条件Logistic回归分析影响血糖的危险因素。选择20只体重在150-200g之间的健康雄性SD大鼠,随机选择15只给予高脂饲料喂养,余下5只给予正常饲料饲养。8周后,给予15只高脂饲养大鼠禁食12小时(过夜禁食,不禁水),通过腹腔注射链脲佐菌素制备糖尿病大鼠模型,5只健康鼠作为对照组给予注射相当体积的柠檬酸缓冲液。结果,11只成2型糖尿病模型,死亡3只,1只未成模。将糖尿病大鼠随机分为两组,一组给予Ang 1-7(576 ng/d)药泵干预,为Ang 1-7组,5只。另外一组泵入相当体积生理盐水,为糖尿病组,6只;2周后处死所有大鼠,留取肾脏标本。留取血、尿标本检测血肌酐和24小时尿蛋白。Western印迹和实时荧光定量PCR检测分子伴侣葡萄糖调节蛋白78(GRP78),增强子结合蛋白同源蛋白(CHOP)和转录活性因子4(ATF4)水平。Western印迹和免疫组化检测大鼠肾组织凋亡情况。结果:不同血糖水平人群研究结果显示,糖耐量异常组和糖尿病组甘油三酯、ACE2、Ang 1-7水平与对照组相比明显升高;糖尿病组较对照组组明显升高(p0.017),(TG:3.4±0.9 v.s 1.5±0.8,ACE2:28.6±4.7 v.s 21.8±4.2,Ang 1-7:14.4±4.2 v.s 11.0±3.5);进一步多因素非条件Logistic回归分析显示,ACE2和Ang 1-7是空腹血糖的独立危险因素。Western印迹结果显示,糖尿病组大鼠肾脏组织的内质网应激标志蛋白(GRP78、ATF4和CHOP)水平明显升高,与正常组相比,其差异有统计学意义(p0.05);与糖尿病组相比,Ang 1-7组网应激标志蛋白水平明显下降,其差异有统计学意义(p0.05)。Western印迹和免疫组化显示Ang 1-7组肾组织caspase 3的表达较糖尿病组明显降低。结论初诊糖尿病患者Ang 1-7水平保护性升高,Ang 1-7可抑制糖尿病大鼠肾脏内质网应激和凋亡。
[Abstract]:Objective: To study 1. patients with different glucose tolerance status of angiotensin 1-7 (Ang 1-7), angiotensin converting enzyme (ACE) level of 2, to explore its role in diabetes and its complications in the preparation of.2. diabetic nephropathy model, to observe the effect of renal lesion in diabetic rats and evaluate Ang 1-7 on diabetic rat kidney endoplasmic reticulum stress and apoptosis. Methods: from December 2014 to the seaside of Tianjin City People's Hospital of 95 medical staff as the object of study, all subjects underwent an oral glucose tolerance test, diagnostic criteria of diabetes with type 2 diabetes prevention guide "China type 2010>2: fasting blood glucose than 7.0mmol/L, postprandial 11.1mmol/L. blood glucose in patients with abnormal blood glucose greater than including impaired fasting glucose (fasting glucose between 6.1~7.0 and mmol/L) impaired postprandial glucose regulation (2 h postprandial blood glucose between 7.8~11.1 mmol/L) patients. According to the blood sugar level in normal group (n=31), Abnormal glucose tolerance group (n=32) and diabetes group (n=32). The detection of body weight, waist circumference, blood pressure, liver, renal function, blood lipid level, Ang 1-7, angiotensin-converting enzyme 2 (ACE) and other indicators, evaluation of three groups of patients with Ang 1-7 level and the relationship with blood glucose, further application of multivariate non conditional Logistic regression analysis of risk factors affecting blood glucose. Healthy male SD rats of 20 body weight between 150-200g, randomly selected 15 rats were given high fat diet, the remaining 5 were given.8 weeks after feeding normal diet, given only 15 in high fat fed rats were fasted for 12 hours (overnight fasting, but not water). The model of diabetes was induced by intraperitoneal injection of streptozotocin, 5 healthy rats as citric acid buffer control group was injected with the equal volume. As a result, 11 type 2 diabetes model, 3 rats died, 1 is not the norm. The diabetic rats were randomly divided into two groups, one Ang group received 1-7 (5 76 ng/d) drug pump intervention, Ang 1-7 group, 5. Another group of equal volume of saline infusion, diabetes group, 6; 2 weeks later, all rats were sacrificed. The kidneys were performed. Blood glucose detection, detection of molecular chaperones of serum creatinine and urine samples of 24 hours urinary protein.Western Western blotting and real-time fluorescent quantitative PCR regulated protein 78 (GRP78), enhancer binding protein homologous protein (CHOP) and transcription factor 4 (ATF4) level of.Western blot and immunohistochemical detection of apoptosis in rat renal tissue. Results: the results showed that people with different blood glucose levels, abnormal glucose tolerance group and diabetes group triglyceride. ACE2, Ang 1-7 levels compared with the control group increased significantly; the diabetic group was significantly increased compared with the control group (p0.017), (TG:3.4 + 0.9 v.s 1.5 + 0.8, ACE2:28.6 + 4.7 v.s 21.8 + 4.2 Ang 1-7:14.4 + 4.2 v.s 11 + 3.5); multivariate non conditional Logistic regression Analysis showed that ACE2 and Ang are 1-7 independent risk factors for fasting blood glucose.Western blot showed that the endoplasmic reticulum stress protein markers in kidney of diabetic rats (GRP78, ATF4 and CHOP) levels were significantly increased, compared with the normal group, the difference was statistically significant (P0.05); compared with the diabetic group, Ang 1-7 network stress marker protein levels were significantly decreased, the difference was statistically significant (P0.05).Western blotting and immunohistochemistry showed that the expression of Ang in renal tissue of 1-7 groups of caspase 3 were significantly lower than those of diabetes mellitus group. Conclusion the Ang patients with newly diagnosed 1-7 diabetes increased the level of protection, Ang 1-7 can inhibit the kidney of diabetic rats on endoplasmic reticulum stress and apoptosis.

【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.2;R692.9

【参考文献】