【摘要】：研究目的：觀察Exendin-4干預(yù)對(duì)糖尿病腎病(DN)大鼠腎組織HO-1.NT蛋白和mRNA表達(dá)的影響,以闡明氧化應(yīng)激機(jī)制是否參與Exendin-4保護(hù)DN大鼠腎臟的作用。 研究方法： 1.實(shí)驗(yàn)設(shè)計(jì)：高糖高脂飼料結(jié)合鏈脲佐菌素(40mg/kg)建立DN大鼠模型,將造模成功大鼠隨機(jī)分為DN組、低劑量干預(yù)組和高劑量干預(yù)組,并與正常組大鼠比較。建模成功后次日,低劑量干預(yù)組皮下注射4μg/(kg·d)的Exendin-4,高劑量干預(yù)組給予20μg/(kg·d)的Exendin-4,連續(xù)給藥六周。免疫組化法觀察HO-1以及NT蛋白表達(dá)的變化,RT-PCR法觀察HO-1和NT mRNA表達(dá)的變化。 2.統(tǒng)計(jì)分析：采用單因素方差分析和多重比較探討各組間HO-1、NT蛋白和mRNA表達(dá)的變化。 研究結(jié)果： 1.大鼠一般狀況：正常組大鼠自由飲食和攝水,DN大鼠精神萎靡、活動(dòng)減少以及體重減輕。 2.免疫組化染色：HO-1蛋白在正常組、Exendin-4低劑量干預(yù)組和高劑量干預(yù)組腎小管均有表達(dá),但在DN組表達(dá)較弱或幾乎不表達(dá)。其平均灰度值依次為：正常組(11.0±2.2),DN組(0.5±0.1),低劑量干預(yù)組(2.7±0.6),高劑量干預(yù)組(6.5±1.3)；各組間差異具有統(tǒng)計(jì)學(xué)意義(F=99.2,P0.001)。NT蛋白在正常組、Exendin-4低劑量干預(yù)組和高劑量干預(yù)組腎小管表達(dá)較弱,但在DN組表達(dá)顯著提升。其平均灰度值依次為：正常組(0.5±0.1),DN組(5.9±1.7),低劑量干預(yù)組(2.9±0.5),高劑量干預(yù)組(1.7±0.3)；各組間差異具有統(tǒng)計(jì)學(xué)意義(F=55.4,P0.001)。 3. RT-PCR法：HO-1mRNA在正常組和Exendin-4高劑量干預(yù)組顯著表達(dá),且二者表達(dá)量接近；在DN組表達(dá)較弱或幾乎不表達(dá),經(jīng)Exendin-4低劑量干預(yù)后表達(dá)提升,且以Exendin-4高劑量干預(yù)后提升顯著。其2-△△CT值分別為：正常組(32.1±5.4),DN組(4.1±0.7),低劑量干預(yù)組(7.2±0.9),低劑量干預(yù)組(33.0±6.3)；各組間差異具有統(tǒng)計(jì)學(xué)意義(F=110.8,P0.001)。NT mRNA在正常組腎臟組織中輕微表達(dá),在DN組腎臟組織中顯著表達(dá),經(jīng)Exendin-4低劑量干預(yù)后NT mRNA降低,且以Exendin-4高劑量干預(yù)后降低顯著。其2-△△CT值分別為：正常組(1.0±0.2),DN組(10.4±1.6),低劑量干預(yù)組(6.1±1.5),高劑量干預(yù)組(3.2±0.6)；各組間差異具有統(tǒng)計(jì)學(xué)意義(F=104.3,P0.001)。 結(jié)論： 1.DN退化HO-1在大鼠腎臟組織的表達(dá),經(jīng)Exendin-4干預(yù)后表達(dá)顯著提升；DN強(qiáng)化NT在大鼠腎臟組織的表達(dá),經(jīng)Exendin-4干預(yù)后表達(dá)顯著降低。 2.氧化應(yīng)激機(jī)制可能參與Exendin-4保護(hù)DN大鼠腎臟的作用。
[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 鹵2.2) n group (0.5 鹵0.1), low dose group (2.7 鹵0.6) and high dose group (6.5 鹵1.3). But in DN group, the expression increased significantly. The average gray value was (0.5 鹵0.1) in normal group (5.9 鹵1.7), (2.9 鹵0.5) in low-dose intervention group, and (1.7 鹵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 鹵5.4) group (4.1 鹵0.7), low dose group (7.2 鹵0.9), low dose group (33.0 鹵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 鹵0.2) group (10.4 鹵1.6), low dose group (6.1 鹵1.5), high dose group (3.2 鹵0.6), the difference was statistically significant (FF104.3 鹵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.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R587.2;R692

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 郝紹伯;蘭麗珍;;GLP-1類似物Exendin-4對(duì)糖尿病腎病大鼠腎臟SOD、8-OHdG的影響[J];中國(guó)醫(yī)藥指南;2012年17期

2 孔慧平;蘭麗珍;;Exendin-4干預(yù)對(duì)糖尿病腎病大鼠腎臟TGF-β_1mRNA和蛋白表達(dá)的影響[J];中西醫(yī)結(jié)合心腦血管病雜志;2012年02期

3 Vandana Saini;;Molecular mechanisms of insulin resistance in type 2 diabetes mellitus[J];World Journal of Diabetes;2010年03期

，

本文編號(hào)：2136515