过表达维A酸受体α减轻大鼠肾间质纤维化的机制  

The mechanism of overexpression of retinoic acid receptor alpha in attenuating renal interstitial fibrosis in rats

作　　者：余雪云 覃远汉[1] 蒋玲[1] 韦露明[1] 潘竞 席志杨 Yu Xueyun;Qin Yuanhan;Jiang Ling;Wei Luming;Pan Jing;Xi Zhiyang(Department of Pediatrics, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China)

机构地区：[1]广西医科大学第一附属医院儿科,南宁530021

出　　处：《中华实用儿科临床杂志》2019年第5期341-346,共6页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金(81360115).

摘　　要：目的探讨过表达维A酸受体α(RARα)基因减轻大鼠肾间质纤维化(RIF)的分子机制。方法将40只6周龄雄性SD大鼠按随机数字表法分为4组:假手术组、模型组、阴性对照组、转染组,每组10只。模型组大鼠分离并双重结扎左侧输尿管;假手术组大鼠不结扎输尿管;转染组和阴性对照组大鼠分别在建模的基础上转染携带过表达RARα基因的腺相关病毒和阴性对照病毒。2周末处死大鼠,取左肾组织进行病理学检查并计算RIF指数。采用免疫组织化学法检测肾组织Ⅳ型胶原蛋白(Col-Ⅳ)和纤维连接蛋白(FN)的表达,采用实时荧光定量PCR法和Western blot法检测肾组织RARα、抗增殖蛋白(PHB)、转化生长因子-β1(TGF-β1)的mRNA和蛋白表达。结果1.与假手术组相比,模型组大鼠RIF指数显著升高,差异有统计学意义(22.81±2.43比2.34±0.55,q=24.94,P<0.05);模型组和阴性对照组大鼠RIF指数相比,差异无统计学意义(22.81±2.43比22.26±3.43,q=0.67,P>0.05),而转染组大鼠RIF指数显著降低,差异有统计学意义(14.06±2.99比22.81±2.43,q=10.66,P<0.05)。2.与假手术组相比,模型组大鼠肾组织RARα、PHB的mRNA和蛋白表达均显著降低,而TGF-β1的mRNA及蛋白表达,Col-Ⅳ和FN的表达均显著升高,差异均有统计学意义(mRNA:0.43±0.17比1.00±0.00,0.34±0.08比1.00±0.00,2.97±0.54比1.00±0.00,均P<0.05;蛋白:0.25±0.10比0.51±0.06,0.24±0.07比0.58±0.04,0.59±0.09比0.33±0.06,16.01±0.87比8.79±0.39,14.64±0.32比9.36±0.59,均P<0.05);模型组和阴性对照组大鼠肾组织RARα、PHB、TGF-β1的mRNA及蛋白表达,Col-Ⅳ和FN的表达比较,差异均无统计学意义(均P>0.05);与模型组相比,转染组大鼠肾组织RARα、PHB的mRNA和蛋白表达均显著升高,而TGF-β1的mRNA及蛋白表达、Col-Ⅳ和FN的表达均显著降低,差异均有统计学意义(mRNA:0.86±0.07比0.43±0.17,0.89±0.11比0.34±0.08,1.65±0.28比2.97±0.54,均P<0.05;蛋白:0.40±0.07比0.25±0.10,0.45±0.10比0Objective To study the mechanism of overexpression of retinoic acid receptor alpha (RARα) in attenuating renal interstitial fibrosis (RIF) in rats. Methods Forty 6-week-old male SD rats were randomly divided into 4 groups: sham operation group, model group, negative control group and transfection group, with 10 rats in each group.Rats in model group were separated and double ligated with left ureter;rats in sham operation group were not ligated with ureter;rats in transfection group and negative control group were transfected with adeno-associated virus and negative control virus carrying RARα gene on the basis of model group, respectively.All rats were sacrificed 2 weeks later.Left kidney tissues were taken for pathological examination and RIF index was calculated.The expression of collagen Ⅳ(Col-Ⅳ) and fibronectin (FN) in renal tissue was detected by using immunohistochemistry.The expressions of RARα, prohibitin (PHB) and transforming growth factor-beta 1 (TGF-β1) in renal tissue were detected by using real-time fluorescence quantitative polymerase chain reaction(RT-qPCR) and Western blot. Results (1)Compared with sham operation group, the RIF index was significantly increased in model group (22.81±2.43 vs.2.34±0.55, q=24.94, P<0.05);compared with model group, the RIF index was not of significant difference in negative control group (22.81±0.43 vs.22.26±3.43, q=0.67, P>0.05), however it significantly decreased in transfection group (14.06±2.99 vs.22.81±2.43, q=10.66, P<0.05).(2)Compared with sham operation group, the mRNA and protein expressions of RARα, PHB significantly decreased in model group, but TGF-β1 mRNA and protein, Col-Ⅳ and FN protein expression significantly increased in model group(mRNA: 0.43±0.17 vs.1.00±0.00, 0.34±0.08 vs.1.00±0.00, 2.97±0.54 vs.1.00±0.00, all P<0.05;protein: 0.25±0.10 vs.0.51±0.06, 0.24±0.07 vs.0.58±0.04, 0.59±0.09 vs.0.33±0.06, 16.01±0.87 vs.8.79±0.39, 14.64±0.32 vs.9.36±0.59, all P<0.05);compared with model group, the mRNA and protein express

关 键 词：维A酸受体α 抗增殖蛋白 肾间质纤维化 转染 

分 类 号：R726.9[医药卫生—儿科]