肾纤康通过调节LncRNA5318介导的TGF-β/Smad信号通路抗肾纤维化作用机制研究

Mechanism of Shenxiankang in Intervening Renal Fibrosis Rats Through Egulating LncRNA5318-mediated TGF-β/Smad Signal Pathway

目的:基于LncRNA5318介导的TGF-β/Smad信号通路,探讨肾纤康对肾纤维化大鼠的保护作用及机制。方法:采用单侧输尿管梗阻(unilateral ureteral obstruction,UUO)肾间质纤维化模型。将25只大鼠随机分为5组,每组5只,即假手术组、模型组、贝那普利组、肾纤康低剂量组、高剂量组。各给药组给予相应药物灌胃,假手术组和模型组给予等体积生理盐水灌胃,每日1次,连续2周。HE和Masson染色观察各组大鼠肾组织病理改变,计算肾小管间质损伤指数及肾纤维化相对面积;Western blot法检测肾组织α平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、转化生长因子β1(transforming growth factor-β1,TGF-β1)、p-Smad2、Smad2、p-Smad3、Smad3、Smad7蛋白的表达;qPCR法检测肾组织LncRNA5318的表达。结果:与假手术组相比,模型组大鼠肾脏出现明显损伤及纤维化;与模型组相比,贝那普利和肾纤康治疗组肾小管间质损伤指数及肾纤维化相对面积明显下降(P<0.05)。Western blot检测提示与假手术组相比,模型组α-SMA、TGF-β1、p-Smad2、p-Smad3的表达均明显增强,而Smad7表达明显减弱(P<0.05);贝那普利和肾纤康治疗后α-SMA、TGF-β1、p-Smad2、p-Smad3的表达明显减少,Smad7的表达明显增多(P<0.05)。qPCR检测提示与假手术组相比,模型组LncRNA5318的表达明显增多(P<0.05);贝那普利和肾纤康治疗后LncRNA5318的表达明显减少(P<0.05)。结论:肾纤康可以延缓肾纤维化,疗效与贝那普利相当,作用机制可能与通过调节LncRNA 5318介导的TGF-β/Smad信号通路有关。

Objective:To investigate the protective effect and mechanisms of Shenxiankang on renal fibrosis rats based on TGF-β1/Smad pathway mediated by LncRNA5318.Methods:Models of renal interstitial fibrosis in rats were established with unilateral urethral obstruction(UUO).Twenty-five rats were randomly divided into five groups:sham operation group,model group,benazepril group,shenxiankang low-dose and high-dose group.Corresponding drugs were given to intervene by gastric gavage in administration groups,and the same volume of normal saline was given by gastric gavage in sham surgery group and model group,Once a day for 2 consecutive weeks.Pathological changes of the renal tissue were observed by HE and Masson staining,the renal tubulointerstitial injury index and relative area of renal fibrosis were calculated.The protein expression levels ofα-smooth muscle actin(α-SMA),transforming growth factor-β1(TGF-β1),p-Smad2,Smad2,p-Smad3,Smad3 and Smad7 were detected by Western blot.The expression level of LncRNA5318 in kidney tissue was detected by real-time quantitative qPCR.Results:Compared with sham surgery group,obvious kidney injury and fibrosis were observed in model group.Compared with the model group,tubulointerstitial injury index and relative area of renal fibrosis in benazepril and shenxiankang treatment groups were significantly decreased(P<0.05).Compared with the sham operation group,the protein expression levels ofα-SMA,TGF-β1,p-Smad2 and p-Smad3 in the model group were significantly increased(P<0.05),while the expression levels of Smad7 was significantly decreased(P<0.05).Benazepril and shenxiankang could reduce the protein expression levels ofα-SMA,TGF-β1,p-Smad2 and p-Smad3 and mprove the expression level of Smad7(P<0.05).Compared with the sham operation group,the expression level of LncRNA5318 in the model group was significantly increased(P<0.05),Benazepril and shenxiankang could reduce the expression level of LncRNA5318(P<0.05).Conclusion:Shenxiankang can alleviate renal fibrosis,and its efficacy is comparable to that of benazepril.The mechanism may be related to the regulation of TGF-β/Smad signal pathway mediated by LncRNA5318.