Exendin-4干预对2型糖尿病肾病大鼠肾组织HO-1及NT表达的影响
发布时间：2018-07-21 19:00
【摘要】：研究目的：观察Exendin-4干预对糖尿病肾病(DN)大鼠肾组织HO-1.NT蛋白和mRNA表达的影响,以阐明氧化应激机制是否参与Exendin-4保护DN大鼠肾脏的作用。 研究方法： 1.实验设计：高糖高脂饲料结合链脲佐菌素(40mg/kg)建立DN大鼠模型,将造模成功大鼠随机分为DN组、低剂量干预组和高剂量干预组,并与正常组大鼠比较。建模成功后次日,低剂量干预组皮下注射4μg/(kg·d)的Exendin-4,高剂量干预组给予20μg/(kg·d)的Exendin-4,连续给药六周。免疫组化法观察HO-1以及NT蛋白表达的变化,RT-PCR法观察HO-1和NT mRNA表达的变化。 2.统计分析：采用单因素方差分析和多重比较探讨各组间HO-1、NT蛋白和mRNA表达的变化。 研究结果： 1.大鼠一般状况：正常组大鼠自由饮食和摄水,DN大鼠精神萎靡、活动减少以及体重减轻。 2.免疫组化染色：HO-1蛋白在正常组、Exendin-4低剂量干预组和高剂量干预组肾小管均有表达,但在DN组表达较弱或几乎不表达。其平均灰度值依次为：正常组(11.0±2.2),DN组(0.5±0.1),低剂量干预组(2.7±0.6),高剂量干预组(6.5±1.3)；各组间差异具有统计学意义(F=99.2,P0.001)。NT蛋白在正常组、Exendin-4低剂量干预组和高剂量干预组肾小管表达较弱,但在DN组表达显著提升。其平均灰度值依次为：正常组(0.5±0.1),DN组(5.9±1.7),低剂量干预组(2.9±0.5),高剂量干预组(1.7±0.3)；各组间差异具有统计学意义(F=55.4,P0.001)。 3. RT-PCR法：HO-1mRNA在正常组和Exendin-4高剂量干预组显著表达,且二者表达量接近；在DN组表达较弱或几乎不表达,经Exendin-4低剂量干预后表达提升,且以Exendin-4高剂量干预后提升显著。其2-△△CT值分别为：正常组(32.1±5.4),DN组(4.1±0.7),低剂量干预组(7.2±0.9),低剂量干预组(33.0±6.3)；各组间差异具有统计学意义(F=110.8,P0.001)。NT mRNA在正常组肾脏组织中轻微表达,在DN组肾脏组织中显著表达,经Exendin-4低剂量干预后NT mRNA降低,且以Exendin-4高剂量干预后降低显著。其2-△△CT值分别为：正常组(1.0±0.2),DN组(10.4±1.6),低剂量干预组(6.1±1.5),高剂量干预组(3.2±0.6)；各组间差异具有统计学意义(F=104.3,P0.001)。 结论： 1.DN退化HO-1在大鼠肾脏组织的表达,经Exendin-4干预后表达显著提升；DN强化NT在大鼠肾脏组织的表达,经Exendin-4干预后表达显著降低。 2.氧化应激机制可能参与Exendin-4保护DN大鼠肾脏的作用。
[Abstract]:Aim: to investigate the effects of Exendin-4 intervention on the expression of HO-1.NT protein and mRNA in diabetic nephropathy (DN) rats, and to elucidate whether the oxidative stress mechanism is involved in the protective effect of Exendin-4 on the kidney of diabetic nephropathy (DN) rats. Methods: 1. Design: diabetic rats were established with high glucose and high fat diet combined with streptozotocin (40mg/kg). The rats were randomly divided into DN group, low dose intervention group and high dose intervention group, and compared with normal rats. The next day after modeling, the low dose group was subcutaneously injected with Exendin-4 of 4 渭 g / (kg d), and the high dose group was given 20 渭 g / (kg d) of Exendin-4 for 6 weeks. The changes of HO-1 and NT protein expression were observed by immunohistochemical method and the changes of HO-1 and NT mRNA expression by RT-PCR. 2. Statistical analysis: univariate ANOVA and multiple comparisons were used to investigate the changes of HO-1NT protein and mRNA expression in each group. Results: 1. General status of rats: normal group of rats fed free diet and water intake of DN rats mental retardation, loss of activity and weight loss. 2. Immunohistochemical staining showed that: HO-1 protein was expressed in the renal tubules of the normal control group and the high dose group, but the expression of the protein was weak or almost no in the DN group. The average gray value was (11.0 卤2.2) n group (0.5 卤0.1), low dose group (2.7 卤0.6) and high dose group (6.5 卤1.3). But in DN group, the expression increased significantly. The average gray value was (0.5 卤0.1) in normal group (5.9 卤1.7), (2.9 卤0.5) in low-dose intervention group, and (1.7 卤0.3) in high-dose intervention group, and the difference was statistically significant (F _ (55.4) P _ (0.001). RT-PCR: HO-1 mRNA was significantly expressed in normal group and Exendin-4 high-dose intervention group, and the expression of HO-1 mRNA was similar to that in DN group, but in DN group, the expression of HO-1 mRNA was increased after low dose Exendin-4 intervention, and increased significantly after high dose Exendin-4 intervention. The 2CT values were: normal group (32.1 卤5.4) group (4.1 卤0.7), low dose group (7.2 卤0.9), low dose group (33.0 卤6.3). NT mRNA decreased after low dose of Exendin-4, and decreased significantly after high dose of Exendin-4. The values of 2-CT were: normal group (1.0 卤0.2) group (10.4 卤1.6), low dose group (6.1 卤1.5), high dose group (3.2 卤0.6), the difference was statistically significant (FF104.3 卤P0.001). Conclusion: 1. The expression of degenerative HO-1 in the kidney of rats was significantly increased after the intervention of Exendin-4, and the expression of NT was significantly decreased after the intervention of Exendin-4. 2. The mechanism of oxidative stress may be involved in the protective effect of Exendin-4 on the kidney of DN rats.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R587.2;R692
本文编号：2136515
[Abstract]:Aim: to investigate the effects of Exendin-4 intervention on the expression of HO-1.NT protein and mRNA in diabetic nephropathy (DN) rats, and to elucidate whether the oxidative stress mechanism is involved in the protective effect of Exendin-4 on the kidney of diabetic nephropathy (DN) rats. Methods: 1. Design: diabetic rats were established with high glucose and high fat diet combined with streptozotocin (40mg/kg). The rats were randomly divided into DN group, low dose intervention group and high dose intervention group, and compared with normal rats. The next day after modeling, the low dose group was subcutaneously injected with Exendin-4 of 4 渭 g / (kg d), and the high dose group was given 20 渭 g / (kg d) of Exendin-4 for 6 weeks. The changes of HO-1 and NT protein expression were observed by immunohistochemical method and the changes of HO-1 and NT mRNA expression by RT-PCR. 2. Statistical analysis: univariate ANOVA and multiple comparisons were used to investigate the changes of HO-1NT protein and mRNA expression in each group. Results: 1. General status of rats: normal group of rats fed free diet and water intake of DN rats mental retardation, loss of activity and weight loss. 2. Immunohistochemical staining showed that: HO-1 protein was expressed in the renal tubules of the normal control group and the high dose group, but the expression of the protein was weak or almost no in the DN group. The average gray value was (11.0 卤2.2) n group (0.5 卤0.1), low dose group (2.7 卤0.6) and high dose group (6.5 卤1.3). But in DN group, the expression increased significantly. The average gray value was (0.5 卤0.1) in normal group (5.9 卤1.7), (2.9 卤0.5) in low-dose intervention group, and (1.7 卤0.3) in high-dose intervention group, and the difference was statistically significant (F _ (55.4) P _ (0.001). RT-PCR: HO-1 mRNA was significantly expressed in normal group and Exendin-4 high-dose intervention group, and the expression of HO-1 mRNA was similar to that in DN group, but in DN group, the expression of HO-1 mRNA was increased after low dose Exendin-4 intervention, and increased significantly after high dose Exendin-4 intervention. The 2CT values were: normal group (32.1 卤5.4) group (4.1 卤0.7), low dose group (7.2 卤0.9), low dose group (33.0 卤6.3). NT mRNA decreased after low dose of Exendin-4, and decreased significantly after high dose of Exendin-4. The values of 2-CT were: normal group (1.0 卤0.2) group (10.4 卤1.6), low dose group (6.1 卤1.5), high dose group (3.2 卤0.6), the difference was statistically significant (FF104.3 卤P0.001). Conclusion: 1. The expression of degenerative HO-1 in the kidney of rats was significantly increased after the intervention of Exendin-4, and the expression of NT was significantly decreased after the intervention of Exendin-4. 2. The mechanism of oxidative stress may be involved in the protective effect of Exendin-4 on the kidney of DN rats.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R587.2;R692
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