不同GLP-1类似物对糖尿病肾病大鼠模型肾脏结构及功能的影响及差异

发布时间：2018-05-07 10:32

本文选题：利拉鲁肽 + 艾塞那肽　；参考：《新乡医学院》2017年硕士论文

【摘要】：背景糖尿病肾病(DN)是临床上常见的糖尿病微血管并发症之一,约30%-40%的糖尿病患者伴有不同程度肾脏的损伤,严重影响患者的生活质量。胰高血糖素样肽1(GLP-1)类似物是近年来运用于临床治疗糖尿病的一种新型药物,通过与其高亲和力受体(GLP-1R)结合,能够促进胰岛素分泌、降低胰高血糖素分泌,同时延缓胃排空、增强饱食感、抑制食欲、增加肝糖原合成、减少肝糖原分解等发挥降低血液中葡萄糖浓度的作用。GLP-1Rs广泛存在于肾脏、心脏、肝脏等身体重要器官。近年来,大量研究表明GLP-1类似物在DN的发病过程中可通过减少炎症反应、抑制氧化应激、改善晚期糖末终基化产物(AGEs)等多个方面起到除降糖作用外的肾脏保护作用。目前已在我国上市应用的GLP-1类似物有利拉鲁肽(liraglutide)和艾塞那肽(exenatide)两种。利拉鲁肽和艾塞那肽都可通过与GLP-1R结合延缓DN的发展,但两者的合成来源、代谢途径均不相同,两者在DN患者中应用的效果是否存在差异尚不清楚。目的探讨验证GLP-1类似物治疗DN的作用机制;对比利拉鲁肽与艾塞那肽在DN发病过程中的对肾脏结构和功能的保护作用是否存在差异。方法健康的雄性SD大鼠50只,完全随机分出正常对照组(10只),另外40只大鼠饲以高糖高脂饲料6周诱导胰岛素抵抗,联合空腹腹腔注射小剂量链脲佐菌素(STZ)破坏胰岛β细胞,形成胰岛素抵抗伴胰岛素分泌相对不足为特征的2型糖尿病,之后3天、7天各测一次晨空腹尾静脉血糖,两次血糖均≥16.9 mmol/L为DN模型成立。成模大鼠32只随机分为模型组(10只)、利拉鲁肽治疗组(11只)和艾塞那肽治疗组(11只)。正常对照组和模型组大鼠予皮下注射生理盐水(10μL/Kg/12h);利拉鲁肽治疗组大鼠予皮下注射利拉鲁肽(0.3 mg/Kg/12h);艾塞那肽治疗组大鼠予皮下注射艾塞那肽(10μL/Kg/12h),共给药8周。第8周末采集大鼠血液标本,肾脏标本。新鲜血液标本离心后收集血清,之后使用全自动生化分析仪测定大鼠BUN和Scr水平;肾脏标本制作成蜡块,采用HE染色和过碘酸雪夫染色(PAS)观察肾脏组织结构变化;免疫组织化学法检测大鼠肾脏组织中TGF-β1和NOX 4的蛋白水平表达并使用Image-pro Plus软件对TGF-β1和NOX 4在肾脏组织中蛋白水平表达进行半定量分析;实时荧光定量PCR(RT-qPCR)定量分析各组肾脏组织中TGF-β1和NOX 4的mRNA水平表达情况。采用SPSS 19.0软件进行统计分析,所有计量资料结果采用均数±标准差(x±s)表示。多组间比较采用单因素方差分析,2组间比较采用t检验,检验水准α=0.05。结果与正常对照组比较,模型组和艾塞那肽治疗组大鼠空腹血糖、BUN、Scr及肾组织中TGF-β1、NOX 4蛋白水平均显著升高(P0.05);利拉鲁肽治疗组大鼠空腹血糖和肾组织中TGF-β1蛋白水平显著升高(P0.05);模型组、利拉鲁肽治疗组和艾塞那肽治疗组大鼠体质量显著降低(P0.05),肾组织形态学均显示肾小球系膜明显增厚,其中模型组增厚最明显,利拉鲁肽治疗组最轻。与模型组比较,利拉鲁肽治疗组和艾塞那肽治疗组大鼠BUN、Scr水平显著降低(P0.05);利拉鲁肽治疗组大鼠肾组织中TGF-β1、NOX 4蛋白水平和mRNA水平显著降低(P0.05);艾塞那肽治疗组大鼠肾组织中TGF-β1、NOX 4蛋白水平和TGF-β1 m RNA水平显著降低(P0.05)。利拉鲁肽治疗组大鼠Scr和肾组织中TGF-β1、NOX 4蛋白水平和m RNA水平显著低于艾塞那肽治疗组(P0.05)。结论1.GLP-1类似物可降低肾脏组织TGF-β1、NOX 4蛋白水平和mRNA水平的表达,降低DN大鼠BUN、Scr水平,延缓大鼠DN的发展;2.DN大鼠短期内使用GLP-1类似物,利拉鲁肽较艾塞那肽能够更好的减少DN模型大鼠肾脏组织中TGF-β1、NOX 4蛋白和mRNA水平的表达,获得更低的BUN、Scr水平。
[Abstract]:Background diabetic nephropathy (DN) is one of the most common diabetic microvascular complications. About 30%-40% diabetic patients with different degrees of kidney injury seriously affect the quality of life. Glucagon like peptide 1 (GLP-1) analogue is a new drug used in the clinical treatment of diabetes in recent years, through its high affinity. Force receptor (GLP-1R) binding, can promote insulin secretion, reduce glucagon secretion, delay gastric emptying, enhance satiety, inhibit appetite, increase liver glycogen synthesis, reduce liver glycogen decomposition, and reduce the concentration of glucose in the blood,.GLP-1Rs widely exists in important organs such as kidney, heart, liver and so on. A large number of studies have shown that GLP-1 analogues can play a role in renal protection by reducing inflammatory response, inhibiting oxidative stress and improving late end glycylation products (AGEs) in the pathogenesis of DN. The GLP-1 analogues in China are currently used in China, including liraglutide and exenat. IDE) two. Alralurin and alisenin can both delay the development of DN by combining with GLP-1R, but the source and metabolic pathways of both are different. The difference in the effect of the two in DN patients is not clear. The purpose is to verify the mechanism of GLP-1 analogues for the treatment of DN, and the pathogenesis of the GLP-1 in the DN. There were differences in the protection of renal structure and function in the process. Methods 50 healthy male SD rats were randomly divided into normal control group (10 rats), and 40 rats were fed with high glucose and high fat diet for 6 weeks to induce insulin resistance, and intraperitoneal injection of small dose streptozotocin (STZ) to destroy islet beta cells and form islets. In type 2 diabetes, which was characterized by relative deficiency of insulin secretion, 3 days after 3 days, every morning caudal vein blood glucose was measured on the 7 day, and two times of blood glucose more than 16.9 mmol/L were established. 32 rats were randomly divided into model group (10), lealaru treatment group (11) and alenenin group (11). Normal control group and model group. Rats were subcutaneously injected with saline (10 L/Kg/12h), and the rats in the learalu peptide group were given subcutaneous injection of Leila Lou (0.3 mg/Kg/12h), and the rats were given the subcutaneous injection of isinanpeptide (10 mu L/Kg/12h) for 8 weeks. The blood specimens of rats were collected at the end of the eighth week, and the kidney specimens were collected. The level of BUN and Scr in rats was measured with an automatic biochemical analyzer. The kidney specimens were made into wax blocks, HE staining and iodate Schiff staining (PAS) were used to observe the changes of renal tissue structure. The protein level of TGF- beta 1 and NOX 4 in the kidney tissues of rats was detected by immunohistochemistry and the Image-pro Plus software was used for the TGF- beta 1 and NOX 4 in the kidney. The expression of protein level in the tissue was analyzed by semi quantitative analysis; real-time quantitative PCR (RT-qPCR) was used to quantitatively analyze the mRNA level expression of TGF- beta 1 and NOX 4 in each group. SPSS 19 software was used for statistical analysis. The results of all measurement data were expressed with mean mean + standard deviation (x + s). A single factor analysis of variance was used in multiple groups, 2 Compared with the normal control group, the results of t test were compared with the normal control group. The level of fasting blood glucose, BUN, Scr, TGF- beta 1 and NOX 4 protein in the model group and the alennisin group were significantly increased (P0.05), and the level of the fasting blood glucose and the TGF- beta 1 protein in the renal tissue was significantly increased (P0.05) in the lealaru treatment group (P0.05); the model was significantly increased (P0.05). Group, the mass of the rats in the Leila Lou treatment group and the alsna peptide group decreased significantly (P0.05). The renal histomorphology showed the glomerular mesangial thickening. The thickening of the model group was the most obvious, and the lealulu treatment group was the lightest group. Compared with the model group, the level of BUN and Scr in the lialalu treatment group and the alsna peptide group were significantly lower (P0.05 TGF- beta 1, NOX 4 protein level and mRNA level in rat renal tissue were significantly decreased (P0.05), and the level of TGF- beta 1, NOX 4 protein and TGF- beta 1 m RNA decreased significantly (P0.05) in the renal tissue of the treatment group of alisin, and the TGF- beta 1 in the Scr and renal tissues of the lealaru treatment group was significantly lower than that of the kidney tissue. In the P0.05. Conclusion 1.GLP-1 analogues can reduce the expression of TGF- beta 1, NOX 4 protein and mRNA level in renal tissue, reduce BUN and Scr level in DN rats and delay the development of DN in rats; 2.DN rats use GLP-1 analogues in the short term, and Liraru peptide can reduce the beta 1 in kidney tissue of DN model rats better. The expression of NOX 4 protein and mRNA level showed a lower level of BUN and Scr.

【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.2;R692.9;R-332

【参考文献】