>
网站首页期刊介绍通知公告编 委 会投稿须知电子期刊广告合作联系我们
最新消息:
MicroRNA-34a-5p在肝纤维化中的表达及作用
作者:朱海东  胡东坡 
单位:商丘医学高等专科学校, 河南 商丘 476100
关键词:肝纤维化 肝星状细胞 MicroRNA-34a-5p I型胶原蛋白 沉默调节蛋白1 分泌型糖蛋白-3α 分泌型糖蛋白-5α β-波链蛋白 大鼠 
分类号:R575.2;R33-3
出版年·卷·期(页码):2019·38·第五期(859-865)
摘要:

目的:探讨大鼠肝纤维化组织及活化肝星状细胞中MicroRNA-34a-5p水平及其在肝纤维化中的作用。方法:建立肝纤维化大鼠模型,转化生长因子-β1(TGF-β1)活化肝星状细胞,采用HE染色观察肝脏组织形态学变化,采用RT-PCR测定大鼠肝组织和活化肝星状细胞中α-平滑肌肌动蛋白(α-SMA)和MicroRNA-34a-5p水平。将肝纤维化模型大鼠根据随机数字表法分为空白对照组、阴性转染对照组和MicroRNA-34a-5p组;将TGF-β1活化的HSC-T6细胞分为空白对照组、阴性转染对照组和MicroRNA-34a-5p组。采用RT-PCR和蛋白质印迹法测定3组大鼠肝组织和肝星状细胞中α-SMA、I型胶原蛋白(collagenI)、沉默调节蛋白1(SIRT1)、分泌型糖蛋白-3α(Wnt-3α)、分泌型糖蛋白-5α(Wnt-5α)和β-波链蛋白(β-catenin)的水平。结果:与对照组比较,肝纤维化组大鼠肝组织和活化肝星状细胞中α-SMA水平升高(P<0.05),MicroRNA-34a-5p水平降低(P<0.05)。与空白对照组和阴性转染对照组比较,MicroRNA-34a-5p转染组大鼠肝组织和活化肝星状细胞中MicroRNA-34a-5p水平升高(P<0.05),而α-SMA、collagenI、SIRT1、Wnt-3α、Wnt-5α和β-catenin的mRNA和蛋白水平均下降(P<0.05),空白对照组和阴性转染对照组间大鼠肝组织和活化肝星状细胞中各指标差异无统计学意义(P>0.05)。结论:肝纤维化组织及活化肝星状细胞中MicroRNA-34a-5p水平降低,可能通过靶向SIRT1而抑制Wnt/β-catenin信号通路,从而发挥抗肝纤维化的作用。

Objective: To investigate the level of MicroRNA-34a-5p in rat liver fibrosis and activated hepatic stellate cells,and to clarify its role in hepatic fibrosis. Methods: The rats model of hepatic fibrosis were established.The hepatic stellate cells were activated by transforming growth factor-β1(TGF-β1). Morphological changes of liver tissue were observed by hematoxylin-eosin (HE) staining.The levels of α-smooth muscle actin (α-SMA) and MicroRNA-34a-5p in rat liver tissue and activated hepatic stellate cells were determined by reverse transcription-polymerase chain reaction (RT-PCR).Liver fibrosis model rats were divided into blank control group, negative transfection control group(transfected with microRNA-34a-5p mimetic negative control) and MicroRNA-34a-5p group(transfected with microRNA-34a-5p simulated matter) according to random number method;The TGF-β1 activated HSC-T6 cells were divided into blank control group, negative transfection control group and MicroRNA-34a-5p group.The α-SMA, collagen I, silencing regulator 1(SIRT1), secreted glycoprotein-3α (Wnt-3α), secreted glycoprotein-5α (Wnt-5α) and β-catenin levels of rat liver tissue and hepatic stellate cells were determined by RT-PCR and Western blotting. Results: Compared with the control group, the levels of α-SMA in the liver fibrosis tissue and activated hepatic stellate cells were increased (P<0.05), and the levels of MicroRNA-34a-5p were decreased (P<0.05).Compared with the blank control group and negative transfection control group, the levels of MicroRNA-34a-5p in the liver tissue and activated hepatic stellate cells of the microRNA-34a-5p group were increased (P<0.05), the α-SMA, collagen I, SIRT1, Wnt-3α, Wnt-5α, β-catenin levels decreased (P<0.05).There was no significant difference in each indicators in the liver tissue and activated hepatic stellate cells between the blank control group and negative transfection control group (P>0.05). Conclusion: MicroRNA-34a-5p level is decreased in hepatic fibrosis and activated hepatic stellate cells. MicroRNA-34a-5p may inhibit Wnt/β-catenin signaling pathway by targeting SIRT1, thereby exerts anti-fibrosis effect.

参考文献:

[1] HUANG Y,DENG X,LIANG J.Modulation of hepatic stellate cells and reversibility of hepatic fibrosis[J].Exp Cell Res,2017,352(2):420-426.
[2] BAE H J,JUNG K H,EUN J W,et al.MicroRNA-221 governs tumor suppressor HDAC6 to potentiate malignant progression of liver cancer[J].J Hepatol,2015,63(2):408-419.
[3] 党树伟,刘明,李国东,等.微RNA-34a在肝癌中的作用研究进展[J].中华肝胆外科杂志,2016,22(7):496-500.
[4] XUE M,LI Y,HU F,et al.High glucose up-regulates microRNA-34a-5p to aggravate fibrosis by targeting SIRT1 in HK-2 cells[J].Biochem Biophys Res Commun,2018,498(1):38-44.
[5] WAN Y,MENG F,WU N,et al.Substance P increases liver fibrosis by differential changes in senescence of cholangiocytes and hepatic stellate cells[J].Hepatology,2017,66(2):528-541.
[6] KIM H S,LEE K S,BAE H J,et al.MicroRNA-31 functions as a tumor suppressor by regulating cell cycle and epithelial-mesenchymal transition regulatory proteins in liver cancer[J].Oncotarget,2015,6(10):8089-8102.
[7] 王伟,方申存,张海涛,等.miRNA-21与器官损伤和纤维化的关系及研究进展[J].东南大学学报(医学版),2018,37(2):335-340.
[8] 石明,仇雪梅,樊红.血浆miRNA检测在肿瘤临床应用的研究进展[J].东南大学学报(医学版),2012,31(1):122-125.
[9] SHEHATA R H,ABDELMONEIM S S,OSMAN O A,et al.Deregulation of miR-34a and Its Chaperon Hsp70 in Hepatitis C virus-Induced Liver Cirrhosis and Hepatocellular Carcinoma Patients[J].Asian Pac J Cancer Prev,2017,18(9):2395-2401.
[10] XIANG Z L,ZHAO X M,ZHANG L,et al.MicroRNA-34a expression levels in serum and intratumoral tissue can predict bone metastasis in patients with hepatocellular carcinoma[J].Oncotarget,2016,7(52):87246-87256.
[11] WANG X P,ZHOU J,HAN M,et al.MicroRNA-34a regulates liver regeneration and the development of liver cancer in rats by targeting Notch signaling pathway[J].Oncotarget,2017,8(8):13264-13276.
[12] 李颖,卢忠心,刘水逸,等.微小RNA-34a在肝癌组织的表达及其临床意义[J].中华实验外科杂志,2016,33(12):2759-2760.
[13] WANG G,ZHAO F,YANG D,et al.Human amniotic epithelial cells regulate osteoblast differentiation through the secretion of TGFβ1 and microRNA-34a-5p[J].Int J Mol Med,2018,41(2):791-799.
[14] LI X Y,WEN J Y,JIA C C,et al.MicroRNA-34a-5p enhances sensitivity to chemotherapy by targeting AXL in hepatocellular carcinoma MHCC-97L cells[J].Oncol Lett,2015,10(5):2691-2698.
[15] 张正威,卢忠心,刘水逸,等.微小RNA-34a靶向X连锁凋亡抑制蛋白基因对肝癌细胞增殖的影响[J].中华实验外科杂志,2015,32(7):1622-1624.
[16] 姚志勇,张超,马鑫,等.microRNA-34a靶向抑制Notch-1基因对膀胱癌J82细胞增殖和迁移的影响[J].中华泌尿外科杂志,2012,33(3):210-214.
[17] ZHU J N,FU Y H,HU Z Q,et al.Activation of miR-34a-5p/Sirt1/p66shc pathway contributes to doxorubicin-induced cardiotoxicity[J].Sci Rep,2017,7(1):11879.
[18] 苏金玲,姜希娟,郭茂娟,等.β连环蛋白在大鼠酒精性肝纤维化中的表达及意义[J].中国老年学杂志,2013,33(13):3097-3098.
[19] ASHMAWY A M,ELGESHY K M,ABDEL-SALAM E T,et al.Crosstalk between liver-related microRNAs and Wnt/β-catenin pathway in hepatocellular carcinoma patients[J].Arab J Gastroenterol,2017,18(3):144-150.
[20] XIANG T,ZHANG S,CHENG N,et al.Oxidored-nitro domain-containing protein 1 promotes liver fibrosis by activating the Wnt/β-catenin signaling pathway in vitro[J].Mol Med Rep,2017,16(4):5050-5054.
[21] 张英,张洪,彭锐,等.青蒿琥酯抑制肝星状细胞microRNA-154/β-catenin治疗肝纤维化的机制研究[J].中国医药导报,2016,13(1):35-38.

服务与反馈:
文章下载】【发表评论】【查看评论】【加入收藏
提示:您还未登录,请登录!点此登录
您是第 405348 位访问者


copyright ©《东南大学学报(医学版)》编辑部
联系电话:025-83272481 83272483
电子邮件:
bjb@pub.seu.edu.cn

苏ICP备09058364