肾间质纤维化动物模型中整合素连接激酶及相关因子的表达

Expression of integrin-linked kinase and associated factors in animal models of renal interstitial fibrosis

背景:在各种原因引起的进展性慢性肾脏病中,均可见到肾间质纤维化现象,其病变程度与慢性肾脏病的预后甚为密切,并可作为预测肾功能恶化的一个十分准确的指标。目的:探讨整合素连接激酶、转化生长因子β1及α-平滑肌肌动蛋白与肾间质纤维化中的关系。设计、时间及地点:随机对照动物实验,2007-03/2008-01在新疆地方与民族高发病省部共建教育部重点实验室完成。材料:成年雄性SD大鼠72只,体质量(200±12)g。方法:72只大鼠随机分为假手术组、模型组和正常对照组,每组24只。模型组大鼠建立肾间质纤维化模型,术后1,3,7,14d麻醉后处死各组大鼠,取梗阻侧肾做苏木精-伊红、PAS及Masson染色。主要观察指标:以免疫组织化学方法检测整合素连接激酶、α-平滑肌肌动蛋白、转化生长因子β1的表达;反转录-聚合酶链反应检测整合素连接激酶的mRNA表达。结果:整合素连接激酶、α-平滑肌肌动蛋白、转化生长因子β1在正常肾组织中表达量极少,但随间质病变的加重,表达量明显增加,与正常组织比差异有非常显著性意义(P<0.001)。其与间质纤维化程度呈正相关(r=0.841,0.892,0.854,P<0.001),在间质中的表达量也成正相关。结论:整合素连接激酶、α-平滑肌肌动蛋白、转化生长因子β1随着间质病变程度的加重而明显增加,说明与肾间质纤维化的形成有密切关系。

BACKGROUND： The phenomenon of renal interstitial fibrosis appears in the progressivity chronicity kidney disease, which has closely relationship with prognosis. The degree renal interstitial fibrosis may be one of the best markers for the diagnosis of renal tubulointerstitial lesions. OBJECTIVE： To explore the correlation of integrin-linked kinase （ILK）, transforming growth factor （TGF）- β1, α-smooth muscle actin （SMA） and renal interstitial fibrosis. DESIGN, TIME AND SETTING： The randomized control experiment of animal was performed at the Key Laboratory of Xinjiang Endemic and Ethnic Diseases Cooperated by Education Ministry with Xinjiang Uighur Autonomous Region between March 2007 and January 2008. MATERIALS： Seventy-two adult, male, Sprague Dawley rats, weighing （200±12） g were selected. METHODS： A total of 72 rats were randomly divided into the sham operation, model and control groups, with 24 animals in each group. Rats in the model group were prepared for tubulointerstitial fibrosis models. All animals were sacrificed at 1, 3, 7, 14 clays after operation and harvested the obstructed kidney for hematoxylin-eosin, PAS and Masson staining. MAIN OUTCOME MEASURES： Expression of ILK,α - SMA, TGF- β1 and mRNA were detected by immunohistochemistry and RT-PCR method. RESULTS： The expression of ILK, α - SMA, TGF- β1 could not be detected in normal renal tissues, but it was significantly increased following the renal tubular interstitial damage, significant statistic difference was assumed at P 〈 0.001. The expression of ILK, α - SMA, TGF- β1 was positively related with tubulointerstitial fibrosis （r=0.84t, 0.892, 0.854, P 〈 0.001）, which also positively correlated with expression in interstitial tissues., CONCLUSION： Expression of ILK, α - SMA, and TGF-β1 are obvious increased with the renal tubulointerstitial lesions in a degree dependent manner, which shows a closely relationship with the formation of interstitial fibrosis.