六味地黄汤对UUO大鼠肾组织β-catenin及EMT的影响

Effect of Liuwei Dihuang decoction onβ-catenin and EMT in kidney tissue of UUO rats

目的探讨六味地黄汤(Liuwei Dihuang decoction,LWDHD)对单侧输尿管梗阻(unilateral ureteric obstrutction,UUO)大鼠肾组织中上皮钙黏蛋白E(E-cadherin)、β-连环蛋白(β-catenin)、α-平滑肌肌动蛋白(α-SMA)的表达、肾脏组织病理变化及上皮-间质转化(epithelial-mesenchymal transformation,EMT)的影响。方法SPF级SD大鼠48只随机分成假手术组(Sham)、模型组(UUO)、LWDHD低、中、高组(LWDHD 3.375、6.75、13.5 g·kg^(-1))、依那普利组(Enalapril)10 mg·kg^(-1),每组8只。采用HE、Masson染色法观察6组大鼠患侧肾组织的病理变化和胶原纤维沉积情况;免疫组化及Western blot检测各组大鼠肾组织中上E-cadherin、β-catenin、α-SMA蛋白的表达情况。结果与Sham组比较,UUO组大鼠患侧肾组织中肾小球硬化,肾小管扩张,间质纤维化明显。β-catenin、α-SMA蛋白表达增加,E-cadherin蛋白表达降低(P<0.05);与UUO组相比,LWDHD组和Enalapril组中大鼠肾组织病理改变和胶原沉积明显改善,β-catenin、α-SMA蛋白表达降低,E-cadherin蛋白表达增加(P<0.05)。结论LWDHD可能通过下调β-catenin的表达,进而抑制EMT,发挥缓解肾纤维化的作用。

Aim To explore the effect of Liuwei Dihuang decoction(LWDHD)on the expression ofβ-catenin,E-cadherin,α-SMA,the pathological changes of renal tissue,and the changes of an epithelial-mesenchymal transformation(EMT)in renal tissue of rats with unilateral ureteral obstruction(UUO).Methods Forty-eight SPF grade SD rats were randomly divided into sham group(Sham),model group(UUO),Liuwei Dihuang decoction low,medium,and high groups(LWDHD 3.375,6.75,13.5 g·kg^(-1))and enalapril group(10 mg·kg^(-1)),with eight rats in each group.HE and Masson staining were used to observe the pathological changes and collagen fiber deposition in the affected renal tissue of the six groups.Immunohistochemistry and Western blot were employed to detect the expression of E-cadherin,β-catenin,andα-SMA in renal tissue of rats in each group.Results Compared with the sham group,the UUO group showed glomerular sclerosis,tubular dilatation,and more severe interstitial fibrosis.The expression ofβ-catenin andα-SMA protein increased,while the expression of E-cadherin protein decreased(P<0.05).Compared with the UUO group,the pathological changes and collagen deposition of renal tissue in the LWDHD group and enalapril group were significantly improved,the protein levels ofβ-catenin andα-SMA decreased,while the protein level of E-cadherin increased(P<0.05).Conclusions LiuWei Dihuang decoction may down-regulateβ-catenin expression,and then inhibit EMT,thus playing a role in alleviating renal fibrosis.