益气活血降浊复方对单侧输尿管梗阻大鼠肾小管上皮细胞转分化影响的实验研究

目的:研究益气活血降浊复方对单侧输尿管梗阻(UUO)大鼠肾组织E-cadherin,α-SMA蛋白表达情况,观察本方对肾小管上皮细胞转分化的影响,进一步探讨其抑制肾间质纤维化的作用机制。方法:132只雄性SD大鼠以单侧输尿管结扎术建立肾间质纤维化动物模型,造模成功后大鼠随机分为正常组,假手术组,模型组,西药组,益气活血降浊复方大、中、小剂量组。根据大鼠体重确定具体给药剂量,益气活血降浊复方大剂量组34.6g.kg-1,中剂量组17.3g.kg-1,小剂量组8.6g.kg-1(分别为人服用量的20,10,5倍)ig给药,1次/天,模型组和假手术组、正常组灌入相应生理盐水。蒙诺10mg.kg-1(相当于成人用量的20倍)。考马斯亮蓝法测定24h尿蛋白定量;检测血清Scr、BUN水平;采用免疫组化方法检测肾组织中E-cadherin,α-SMA蛋白表达。结果:与正常组、假手术组相比,模型组大鼠在第1、2、3周E-cadherin蛋白表达明显减少(P0.01),益气活血降浊复方大、中、小剂量组较正常组、假手术组在各个时间点E-cadherin蛋白表达有所减少,但与模型组比较显著增加(P0.01),福辛普利钠组较模型组表达也显著增加(P0.01),具有统计学意义。中药大剂量组在各个时间点增加E-cadherin蛋白效果最好且与西药组相当,中、小剂量组虽然能增加E-cadherin蛋白表达,但效果逊于大剂量组和西药组。而模型组大鼠第1、2、3周肾间质α-SMA蛋白较正常组、假手术组表达明显增加(P0.01),益气活血降浊复方大、中、小剂量组较正常组、假手术组在各个时间点α-SMA蛋白表达有所增加,但与模型组比较显著减少(P0.01),西药组较模型组表达也显著减少(P0.01),具有统计学意义。中药大剂量组在各个时间点降低α-SMA蛋白效果最好且与西药组相当,中、小剂量组虽然能降低α-SMA蛋白表达,但和大剂量组及西药组存在差异。结论:益气活血降浊复方可能通过抑制α-SMA蛋白表达,上调E-cadherin表达,影响肾小管上皮细胞转分化,减少细胞外基质蛋白的合成,增加其降解,从而减缓肾间质纤维化的发展。

Experimental Study ofYiqihuoxuejiangzhuo Decoction on Renal Tubular Epithelial Cells Transdifferentiation in Unilateral Ureteral Obstruction Rat Model

Objective:This experiment is to study Yiqihuoxuejiangzhuo Decoction on E-cadherin,α-SMA protein expressions,to investigate the effect of Yiqihuoxuejiangzhuo Decoction on renal tubular epithelial cells transdifferentiation,and to illustrate its mechanism of relieving renal interstitial fibrosis.Methods:Unilateral Ureteral Obstruction(UUO) methods were adopted to establish animal models of renal interstitial fibrosis,132 SD rats were randomly divided into normal group,sham operation group,model group,western medicine group,YiqihuoxueJiangzhuo large,medium and low-dose group.Rats in the large,medium and low dose groups were administrated with Yiqihuoxuejiangzhuo Decoction at 34.6,17.3,8.6g·kg-1(equivalent to adult dosage of 20,10,5 respectively),and in western medicine group with 10mg·kg-1 Monopril(equivalent to adult dosage of 20).Coomassie brilliant blue method was used to measure 24h urinary protein;Scr and BUN changes in blood serum were observed.The protein expressions of E-cadherin and α-SMA in kidney tissue were detected respectively by immunohistochemistry.Results:Compared with the normal group,sham operation group,E-cadherin protein expressions in rat model group were significantly reduced at 1,2,3 weeks(P<0.01),E-cadherin protein expressions of Yiqihuoxue jiangzhuo Decoction large,medium,low-dose groups were reduced than that in the normal group,sham operation group at each time point.However,compared with the model group they were significantly increased(P<0.01).Western Fosinopril sodium group were also significantly increased than in model group(P<0.01),with statistical significance.In this experiment,Yiqihuoxuejiangzhuo high dose group increased E-cadherin protein best at each time point,and to be equivalent of western medicine group.The medium and low dose group can also increase the E-cadherin protein,but there are differences with the high-dose group and western medicine group.In contrast,α-SMA protein expressions were reduced in YiqihuoxueJiangzhuo large,medium and low dose groups compared with model groups.Conclusion:Yiqihuoxuejiangzhuo Decoction could block renal tubular epithelial cells transdifferentiation by reducing α-SMA protein expressions and increasing E-cadherin protein expressions in kidney tissue.It might be the mechanisms of Yiqihuoxuejiangzhuo Decoction relieving renal interstitial fibroblast.