文献类型：期刊文献

中文题名：基于RhoA/ROCK通路探讨淫羊藿苷对肾病综合征大鼠的保护机制

英文题名：Protective Mechanism of Icariin on Nephrotic Syndrome Rats Based on RhoA/ROCK Pathway

作者：王新斌[1];戴恩来[1];薛国忠[1];吕娟[1];陈威辛[1]

第一作者：王新斌

机构：[1]甘肃中医药大学中西医结合学院,兰州730000

第一机构：甘肃中医药大学中西医结合学院

年份：2020

卷号：26

期号：11

起止页码：78

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD2019_2020】；

基金：国家自然科学基金项目(81760799);甘肃省自然科学基金项目(1610RJ2A067)。

语种：中文

中文关键词：淫羊藿苷;肾病综合征;大鼠;肾素同源蛋白A(RhoA);Rho相关激酶1(ROCK1);ROCK2

外文关键词：icariin;nephrotic syndrome;rats;renin homologous protein A(RhoA);Rho-related kinase 1(ROCK1);ROCK2

摘要：目的:研究淫羊藿苷对肾病综合征(NS)大鼠肾素同源蛋白A(RhoA)/Rho相关激酶(ROCK)通路的影响和保护机制。方法:实验对象为54只清洁级雄性SD大鼠,随机均分为正常组,模型组,RhoA抑制剂组(Rhosin,40 mg·kg^-1·d^-1)和淫羊藿苷低、中、高剂量组(30,60,120 mg·kg^-1·d^-1)。模型组大鼠尾静脉注射6.5 mg·kg^-1盐酸阿霉素给,诱导大鼠NS发生。造模后腹膜给药,正常组和模型组给予生理盐水2.5 mL·d^-1,抑制剂组和各剂量组分别给对应剂量的Rhosin和淫羊藿苷进行干预。试剂盒检测各组大鼠尿总蛋白(Alb),尿肌酐(Cre)水平,计算尿总蛋白/肌酐值(A/C);透射电镜(TEM)鉴定肾脏超微病理;实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测RhoA,ROCK1,ROCK2 mRNA水平和蛋白表达。结果:TEM结果显示,正常组基底膜完整,足突规整;模型组基底膜损坏严重,足突出现消失、融合严重;淫羊藿苷低剂量组基底膜损伤减轻,足突的数量和密度改善、融合明显;淫羊藿苷中剂量组和抑制剂组,基底膜增厚情况减轻,足突轻度融合;淫羊藿苷高剂量组基底膜结构较完整,足细胞较长、排列较紧密。与正常组比较,模型组大鼠24 h尿蛋白量、尿液中Cre,A/C,肾组织RhoA,ROCK1,ROCK2 mRNA和蛋白水平明显升高(P<0.05)。与模型组比较,抑制剂组和淫羊藿苷低、中、高剂量组24 h尿蛋白量,A/C和肾组织中RhoA,ROCK1,ROCK2 mRNA和蛋白水平,抑制剂组和淫羊藿苷高剂量组尿液中Cre明显降低(P<0.05)。与抑制剂组比较,淫羊藿苷高剂量组24 h尿蛋白量、尿液中Cre和肾组织RhoA蛋白水平明显降低(P<0.05);淫羊藿苷低剂量组24 h尿蛋白量、尿液中Cre,A/C和肾组织RhoA,ROCK1,ROCK2 mRNA和蛋白水平,淫羊藿苷中剂量组肾组织RhoA,ROCK1,ROCK2 mRNA水平明显升高(P<0.05)。与淫羊藿苷低剂量组比较,淫羊藿苷中、高剂量组尿液中Alb,A/C和肾组织RhoA,ROCK1,ROCK2 mRNA和蛋白水平,淫羊藿苷高剂量组尿液中Cre明显降低(P<0.05)。与淫羊藿苷中剂量组比较,淫羊藿苷高剂量组24 h尿蛋白量、尿液中Cre和肾组织RhoA,ROCK1 mRNA和蛋白水平,肾组织ROCK2 mRNA明显降低(P<0.05)。结论:在NS大鼠治疗过程中,淫羊藿苷可能通过影响RhoA/ROCK通路,实现肾小球内皮和足细胞保护作用。
Objective:To study the effect of icariin on renin homologous protein A(RhoA)/Rhorelated kinase(ROCK)pathway in rats with nephrotic syndrome(NS)and its protective mechanism.Method:Totally 54 clean-grade male SD rats were tested and randomly divided into normal group,model group,RhoA inhibitor group(Rhosin,40 mg·kg^-1·d^-1)and three doses of icariin groups(low,medium and high corresponding dose,30,60,120 mg·kg^-1·d^-1).Adriamycin hydrochloride 6.5 mg·kg^-1 was given in tail vein of rats to induce NS model in rats.After the model was established,peritoneal administration was carried out.The normal group and the model group were given saline 2.5 mL·d^-1,and the inhibitor group and all of dose groups were given corresponding doses of Rhosin and icariin for intervention.Total urinary protein(Alb),creatinine(Cre),total urinary protein/creatinine ratio(A/C)kit were detected in rats,ultrastructure of kidney was identified by transmission electron microscopy(TEM),and Real-time fluorescent quantitative polymerase chain reaction(Real-time PCR)and Western blot were used to detect the mRNA and proteins expressions of RhoA,ROCK1,ROCK2.Result:TEM showed that the basement membrane was intact and the foot process was regular in the normal group,in model group,basement membrane was damaged seriously,foot process disappeared,and fusion was serious,in the low-dose group,the basement membrane injury was alleviated,the number and density of foot process were improved,and the fusion was obvious,in the middle-dose group and the inhibitor group,the basement membrane thickening was alleviated,and the foot process was slightly fused,in the high-dose group,the basement membrane structure was more complete,and podocytes were longer and arranged tightly.Compared with the normal group,the levels of Alb,Cre and A/C in urine,and RhoA,ROCK1 and ROCK2 mRNA and protein expressions in kidney tissue of rats of the model group were significantly higher(P<0.05).Compared with model group,the levels of Alb,A/C in urine and RhoA,ROCK1,ROCK2 mRNA and protein expressions in kidney tissue in the inhibitor group and low,medium and high-dose groups,and Cre in urine in inhibitor group and high-dose group decreased significantly(P<0.05).Compared with the inhibitor group,the levels of Alb,Cre in urine and RhoA protein in kidney tissue in the high-dose group were significantly decreased(P<0.05),the levels of Alb,Cre,A/C in urine and RhoA,ROCK1,ROCK2 mRNA and protein expressions in kidney tissue of the low-dose group,and the levels of RhoA,ROCK1 and ROCK2 mRNA expressions in kidney tissue of the middle-dose group were significantly increased(P<0.05).Compared with the low-dose group,the levels of Alb,A/C in urine,and RhoA,ROCK1,ROCK2 mRNA and protein expressions in kidney tissue in the middle and high-dose groups,Cre in urine of the high-dose group were significantly decreased(P<0.05).Compared with the middle-dose group,the levels of Alb,Cre in urine,and RhoA,ROCK1,ROCK2 mRNA and protein expressions,ROCK2 mRNA expression in kidney tissue in the high-dose group were significantly decreased(P<0.05).Conclusion:Icariin may protect glomerular endothelium and podocyte by affecting RhoA/ROCK pathway in the treatment of NS rats.