环状RNA-42398通过调控TGF-β1/Smads信号通路抑制肝星状细胞活化  

circRNA-42398 inhibits activation of hepatic stellate cells via TGF-β1/Smads signaling pathway modulation

作　　者：周玉平 吕雪幼 杨萍[3] 温晋锋[1,2] 王庆领 叶国良 ZHOU Yu-ping;LV Xue-you;YANG Ping;WEN Jin-feng;WANG Qing-ling;YE Guo-liang(Affiliated Hospital of Medical School,Ningbo University,Ningbo 315020,China;Institute of Digestive Disease,Ningbo University,Ningbo 315020,China;Ningbo College of Health Sciences,Ningbo 315100,China;Medical School of Ningbo University,Ningbo 315211,China)

机构地区：[1]宁波大学医学院附属医院,浙江宁波315020 [2]宁波大学消化疾病研究所,浙江宁波315020 [3]宁波卫生职业技术学院,浙江宁波315100 [4]宁波大学医学院,浙江宁波315211

出　　处：《中国病理生理杂志》2020年第4期688-692,共5页Chinese Journal of Pathophysiology

基　　金：浙江省医药卫生科技计划(No.2018KY709);浙江省中医药科技计划(No.2019ZB114);宁波市自然科学基金资助项目(No.2018A610417)。

摘　　要：目的:探讨小鼠环状RNA-42398(mmucirc42398)对肝星状细胞活化的影响及机制是否与调节TGF-β1/Smads信号通路有关。方法:小鼠肝星状细胞株JS1,分成正常对照组、空载体阴性对照组(vector组)和mmucirc42398过表达组(mmucirc42398组),体外构建mmucirc42398过表达载体,通过脂质体瞬时转染法转入JS1细胞,48 h后RT-qPCR检测mmucirc42398的表达变化,PCR产物经一代测序验证其环化位点,Western blot检测α-平滑肌肌动蛋白(α-SMA)、I型胶原(Col I)、转化生长因子β1(TGF-β1)、Smad2、Smad3、p-Smad2和p-Smad3蛋白表达。结果:与vector组相比,mmucirc42398组的mmucirc42398表达增加(P<0.01),PCR产物测序验证了其环化位点,提示mmucirc42398过表达质粒成功转入JS1细胞并高效表达;在JS1细胞中过表达mmucirc42398后,α-SMA和Col I蛋白表达显著降低(P<0.01),TGF-β1、Smad2和Smad3蛋白表达无显著变化(P>0.05),但p-Smad2和p-Smad3蛋白水平显著降低(P<0.01)。结论:mmucirc42398能抑制肝星状细胞的活化,其机制与调控TGF-β1/smads信号通路有关。AIM:To study the effect of mouse circular RNA-42398(mmucirc42398)over-expression on the activation of hepatic stellate cells.METHODS:Mouse hepatic stellate JS1 cells were cultured and randomly divided into control group,vector group and mmucirc42398 over-expression group.mmucirc42398 over-expression plasmid vector was constructed,and then transiently transfected into JS1 cells using Lipofectamine 2000.The cells were collected48 h after transfection.Expression of mmucirc42398 was detected by RT-qPCR.The backsplice site of PCR products was verified by sequencing.The protein levels ofα-smooth muscle actin(α-SMA),collagen type I(Col I),transforming growth factorβ1(TGF-β1),Smad2,Smad3,p-Smad2 and p-Smad3 in the cells were determined by Western blot.RESULTS:RT-qPCR results showed that the expression of mmucirc42398 was significantly increased after mmucirc42398 over-expression vector was transiently transfected into the JS1 cells(P<0.01).The protein expression levels ofα-SMA and Col I were significantly decreased(P<0.01),and the phosphorylation levels of Smad2 and Smad3 were decreased significantly in mmucirc42398 over expression group(P<0.01).However,the protein expression levels of TGF-β1,Smad2 and Smad3 had no significant change(P>0.05).CONCLUSION:mmu-circ-42398 inhibits the activation of hepatic stellate cells via TGF-β1/Smads signaling pathway modulation.

关 键 词：肝纤维化 肝星状细胞 环状RNA TGF-β1/Smads信号通路 

分 类 号：R513.2[医药卫生—内科学] R363.2[医药卫生—临床医学]