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目的观察MicroRNA-21(miRNA-21)在单侧输尿管梗阻(UUO)大鼠肾脏组织中的表达,初步探讨其在转化生长因子-β1(TGF-β1)/Smad3致大鼠肾间质纤维化(RIF)信号通路中的机制。方法选择20只SD雄性大鼠,分为UUO组和假手术组(Sham组),分别于建模后第3天、第7天各采集每组5只大鼠肾脏组织,应用实时荧光定量PCR(Real-time qPCR)方法检测肾脏组织中miRNA-21的表达变化,采用Masson染色、苏木素-伊红(HE)染色和免疫组织化学技术评价肾组织纤维化,使用免疫组织化学检测肾脏组织中TGF-β1、Smad3、Ⅰ型胶原蛋白(Col-Ⅰ)和α平滑肌肌动蛋白(α-SMA)表达情况。结果建模后第3天和第7天UUO组肾脏组织中miRNA-21与Sham组比较均升高(P<0.01),7dUUO组肾组织中miRNA-21与3dUUO组相比升高(P<0.01)。建模后3、7dUUO组Masson染色纤维化评分、TGF-β1、Samd3、α-SMA及Col-Ⅰ蛋白阳性表达面积较Sham组升高(P<0.01),以上指标7dUUO组较3dUUO组比较表达增强(P<0.01)。UUO组肾组织中miRNA-21的表达与肾间质纤维化评分正相关(r=0.888,P<0.01);肾组织miRNA-21与TGF-β1、Smad3、Col-Ⅰ、α-SMA蛋白表达呈正相关(r=0.799、0.849、0.882、0.896,P<0.01)。结论 UUO大鼠肾组织中miRNA-21在建模后表达上调,且间质纤维化程度越重表达越高,TGF-β1/Smad3信号通路可能是通过上调miRNA-21从而介导大鼠肾间质纤维化。
Objective To investigate the expression of microRNA-21 (miRNA-21) in renal tissues of rats with unilateral ureteral obstruction (UUO) and to explore its role in renal interstitial fibrosis induced by transforming growth factor-β1 (TGF-β1) / Smad3 (RIF) signaling pathway. Methods Twenty male Sprague-Dawley rats were randomly divided into UUO group and sham operation group (Sham group). The kidneys of 5 rats in each group were collected on the 3rd day and the 7th day respectively. Real-time quantitative PCR (Real-time qPCR) was used to detect the expression of miRNA-21 in renal tissues. Masson staining, hematoxylin-eosin (HE) staining and immunohistochemistry were used to evaluate the renal fibrosis. Immunohistochemistry was used to detect the expression of TGF- -β1, Smad3, Col-Ⅰ and α-SMA. Results On the 3rd and 7th day after operation, the expression of miRNA-21 in the UUO group was significantly higher than that in the Sham group (P <0.01), and the expression of miRNA-21 in the UUO group was higher than that in the 3UUUO group (P < 0.01). After modeling, Masson staining fibrosis score, positive expression area of TGF-β1, Samd3, α-SMA and Col-Ⅰ protein in UUO group were higher than those in Sham group (P <0.01), especially in 7UUO group Enhanced (P <0.01). The expression of miRNA-21 in renal tissue of UUO group was positively correlated with the score of renal interstitial fibrosis (r = 0.888, P <0.01). The expression of miRNA-21, TGF-β1, Smad3, (R = 0.799,0.849,0.882,0.896, P <0.01). Conclusion The expression of miRNA-21 in UUO rats’ kidney tissue is up-regulated after modeling, and the more severe the interstitial fibrosis is, the higher the TGF-β1 / Smad3 signal pathway may be through the up-regulation of miRNA-21 Fibrosis.