消瘀泄浊饮对单侧输尿管梗阻大鼠肾小管上皮-间质转分化及PAR-3表达的影响

Effect of Xiaoyu Xiezhuo Decoction on Epithelial-mesenchymal Transition and PAR-3 in Unilateral Ureteral Obstruction Rats

[目的]通过检测极性蛋白-3(partitioning defective-3,PAR-3)在单侧输尿管梗阻(unilateral ureteral obstruction,UUO)所导致的肾间质纤维化大鼠中的表达,探讨消瘀泄浊饮对肾小管上皮-间质转分化(epithelial-mesenchymal transition,EMT)的影响。[方法]将40只雄性SD大鼠随机分为假手术组、模型(UUO)组、西药(贝那普利)组和中药(消瘀泄浊饮)组。从术后1 d起,每天分别给予消瘀泄浊饮药液10.4 g/(kg·d)和盐酸贝那普利溶液1 mg/(kg·d)灌胃,假手术组和模型组以等体积0.9%氯化钠注射液灌胃4周。给药结束后分别检测白蛋白(albumin,ALB)、血尿素氮(blood urea nitrogen,BUN)、血肌酐(serum creatinine,Scr)、尿酸(uric acid,UA)水平;取手术侧肾组织分别采用苏木精-伊红(hematoxylin-eosin,HE)染色及Masson染色进行相关病理检测;采用免疫荧光和免疫印迹法分别检测肾组织PAR-3的表达;采用免疫组化检测肾组织E-钙黏蛋白(E-cadherin)及α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)蛋白的表达。[结果]与假手术组比较,模型组、西药组、中药组BUN、Scr、UA水平均增高,差异有统计学意义(均P<0.05);与模型组比较,中药组BUN、Scr、UA水平均降低,差异有统计学意义(均P<0.05);与西药组比较,中药组Scr、UA水平均降低,差异有统计学意义(均P<0.05)。与假手术组比较,模型组大鼠肾小管间质损伤指数和肾间质纤维化相对面积均升高(均P<0.05);与模型组比较,西药组、中药组肾小管间质损伤指数和肾间质纤维化相对面积均降低(均P<0.05)。与假手术组比较,其余各组大鼠肾组织PAR-3水平均下降(均P<0.05);与模型组比较,中药组肾组织PAR-3表达升高(P<0.05)。与假手术组比较,其余各组大鼠肾组织α-SMA表达增加,E-cadherin表达减少(均P<0.05);与模型组比较,西药组、中药组大鼠肾组织α-SMA表达减少,E-cadherin表达增加(均P<0.05)。[结论] UUO大鼠肾组织PAR-3表达减少,消瘀泄浊饮能通过上调肾组织PAR-3表达、阻止肾小管上皮细胞向间充质细胞转分化,起到改善肾间质纤维化的作用。

[Objective] To investigate the effect of Xiaoyu Xiezhuo Decoction on epithelial-mesenchymal transition(EMT) in unilateral ureteral obstruction(UUO) rats with renal interstitial fibrosis by examining the expression of partitioning defective-3(PAR-3). [Methods] Forty male SD rats were randomly divided into sham operation group, model(UUO) group, western medicine(Benazepril) group and Chinese medicine(Xiaoyu Xiezhuo Decoction)group. The day after operation, the Chinese medicine group and western medicine group were given Xiaoyu Xiezhuo Decoction 10.4 g/(kg·d) and Benazepril 1 mg/(kg·d) respectively, the other two groups were given the same amount of 0.9% sodium chloride aqueous solution, and the drugs were administered for a total of 4 weeks. After administration, the levels of albumin(ALB), blood urea nitrogen(BUN), serum creatinine(Scr), uric acid(UA)were detected. The pathological changes in the renal tissues were observed by both hematoxylin-eosin(HE) staining and Masson staining. The expression of PAR-3 in renal tissues was measured by immunofluorescence and western blot. The expressions of E-cadherin and α-smooth muscle actin(α-SMA) in renal tissues were measured by immunohistochemistry. [Results] Compared with sham operation group, the levels of BUN, Scr and UA increased in model group, western medicine group and Chinese medicine group(P<0.05). Compared with model group, the levels of BUN, Scr and UA decreased in Chinese medicine group(P<0.05). Compared with western medicine group, Chinese medicine group had lower levels of Scr and UA(P<0.05). Compared with sham operation group, the renal tubular interstitial injury index and collagen positive area of model group increased( P<0.05). Compared with model group, both the renal tubular interstitial injury index and collagen positive area of western medicine group and Chinese medicine group significantly decreased(P<0.05). Compared with sham operation group, the level of PAR-3 in renal tissues decreased in other groups(P<0.05). Compared with model group, Chinese medicine group had higher PAR-3 level(P<0.05). Compared with sham operation group, the expression of α-SMA increased in renal tissues of other groups, while the expression of E-cadherin decreased(P <0.05). Compared with model group, western medicine group and Chinese medicine group had lower α-SMA level as well as higher expression of E-cadherin(P <0.05). [Conclusion] The expression of PAR-3 in UUO rats decreased significantly. Xiaoyu Xiezhuo Decoction can ameliorate renal fibrosis by intervening the expression of PAR-3 and inhibiting the EMT process.