芪归益肾方通过调控miR-141对UUO小鼠肾脏纤维化的影响  

Effect of Qigui Yishen Decoction on miR-141 regulation in UUO mice with renal fibrosis

作　　者：尹佳琦 魏明刚[1] 黄星星 夏凡[1] 毛叶勤[1] 谢宇峰 YIN Jia-qi;WEI Ming-gang;HUANG Xing-xing;XIA Fan;MAO Ye-qin;XIE Yu-feng(The First Affiliated Hospital of Soochow University,Suzhou 215006,China)

机构地区：[1]苏州大学附属第一医院,江苏苏州215006

出　　处：《中成药》2018年第4期765-770,共6页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金面上项目(81273723;81473633;81673896);国家中医临床研究基地业务建设第二批科研专项课题(JDZX2015096);苏州市科技局应用基础研究项目(SYS201602);江苏省研究生科研与实践创新计划(KYCX17-2003)

摘　　要：目的探讨芪归益肾方(黄芪、当归、川芎、怀牛膝)通过调控微小RNA-141(miR-141)的表达对单侧输尿管梗阻(unilateral ureteral obstruction,UUO)小鼠肾脏纤维化的影响及其机制。方法采用UUO方法建立肾间质纤维化模型。30只雄性Balb/c小鼠随机分为以下5组,假手术组,模型组,洛汀新组(50 g/kg),芪归益肾方高(50 g/kg)、低(10 g/kg)剂量组。术后连续灌胃10 d后将小鼠处死,留取各组小鼠肾组织。肾组织标本用HE染色观察组织形态学改变,免疫组织化学法检测转化生长因子-β(transforming growth factor-β1,TGF-β1)、胶原蛋白IV(collagen IV,Col IV)、基质金属蛋白酶9(matrix metalloprotein-9,MMP-9)表达,实时定量聚合酶链式反应(realtime quantitative polymerase chain reaction PCR,RT-PCR)法检测miR141、TGF-β1表达。结果 HE染色显示除假手术组外,其余各组存在不同程度的肾间质纤维化改变,而药物组病理损伤较模型组减轻(P<0.05);免疫组织化学结果表明,芪归益肾方高、低剂量组和洛汀新组TGF-β1、Col IV表达明显低于模型组,而MMP-9表达则明显高于模型组(P<0.05);RT-PCR结果表明,与模型组比较,芪归益肾方高、低剂量组和洛汀新组miR-141表达明显升高,而TGF-β1表达明显降低(P<0.05)。结论在小鼠UUO模型中,芪归益肾方可能通过调控miR-141表达来影响TGF-β1和MMP-9表达,减少ECM成分的异常积聚,从而抑制肾脏纤维化进展。AIM To investigate the effect and mechanism of Qigui Yishen Decoction(QGYS,Astragali Radix,Angelicae sinensis Radix,Chuanxiong Rhizoma,Achyranthis bidentatae Radix)on regulating the expression of miR-141 in unilateral ureteral obstruction(UUO)mice with renal fibrosis.METHODS Thirty Balb/c male mice randomly divided into sham-operated group(=6),UUO group(71=6),Lotensin(50 g/kg)group{n-6),QGYS high dose(50 g/kg)group(n=6),and QGYS low dose(10 g/kg)group(n=6)were conducted UUO surgery to promote kidney fibrosis except the six mice in the sham operation group.After a successive 10-day medication of QGYS and Lotensin to mice by oral gavage on daily basis,all mice were killed to procure renal tissue to observe its morphology and pathology changes by HE staining.The expressions of TGF-(31,ColIV,and MMP-9 were analyzed by immunohistochemical method,and the expressions of miR141,TGF-(31 were measured by realtime PCR.RESULTS The obviously pathological injuries including renal interstitial fibrosis were identified by HE staining among the groups intervened with UUO,but the variance in the extent due to different administrations of QGYS and Lotensin was noticed as well(P<0.05).As compared to the UUO group,high and low dose QGYS groups and Lotensin group achieved an up-regulated expression of TGF-pl and ColIV,and a down-regulated expression of MMP-9 by immunohistochemistry(P<0.05),and significantly increased mRNA expression of miR-141,and decreased mRNA expression of TGF-pl by real-time PCR(P<0.05).CONCLUSION In UUO mouse models,QGYS gives influence to TGF-(31and MMP-9 through inducing miR-141 expression change to decrease abnormal accumulation of ECM,and thus inhibits the progression of renal fibrosis.

关 键 词：芪归益肾方 肾脏纤维化 微小RNA 细胞外基质 

分 类 号：R285.5[医药卫生—中药学]