lncRNA MALAT1对肝星状细胞活化的调节作用及其机制  

Regulatory effect of lncRNA MALAT1 on activation of hepatic stellate cell and its mechanism

作　　者：徐菱遥 魏书堂[1] 董勇 孙正路 赵俊波 韩大正[1] XU Lingyao;WEI Shutang;DONG Yong;SUN Zhenglu;ZHAO Junbo;HAN Dazheng(Department of Gastroenterology,First Affiliated Hospital,Henan University,Kaifeng 475100,China)

机构地区：[1]河南大学第一附属医院消化病科,河南开封475100

出　　处：《吉林大学学报（医学版）》2023年第3期697-705,共9页Journal of Jilin University：Medicine Edition

基　　金：河南省卫健委医学科技攻关计划项目(LHGJ20190532)。

摘　　要：目的:探讨长链非编码RNA(lncRNA)肺腺癌转移相关转录本1(MALAT1)在肝纤维化进展过程中对肝星状细胞(HSC)活化的调节作用,并阐明其作用机制。方法:收集25例健康志愿者(健康组,n=25)和25例肝纤维化患者[轻度肝纤维化组(n=12)和重度肝纤维化组(n=13)]血清样本。小鼠HSC分为对照组、转化生长因子β1(TGF-β1)组、TGF-β1+si-NC组、TGF-β1+si-MALAT1组、TGF-β1+si-MALAT1+anti-miR-150-5p组和TGF-β1+si-MALAT1+CXCL14组。采用实时荧光定量PCR(RT-qPCR)法检测各组研究对象血清和各组HSC中MALAT1 mRNA、miR-150-5p和CXC趋化因子配体14(CXCL14)mRNA表达水平,Western blotting法检测各组研究对象血清中CXCL14蛋白表达水平和各组HSC中CXCL14、α-平滑肌肌动蛋白(α-SMA)和Ⅰ型胶原蛋白α1(COL1A1)蛋白表达水平,CCK-8法检测各组HSC增殖活性,免疫荧光法检测各组HSC中α-SMA和COL1A1蛋白表达量,双荧光素酶报告系统检测miR-150-5p与MALAT1和CXCL143’-UTR基因的靶向关系。结果:RT-qPCR法检测,与健康组比较,轻度和重度肝纤维化组患者血清中MALAT1 mRNA和CXCL14 mRNA表达水平升高(P<0.05),miR-150-5p表达水平降低(P<0.05);与轻度肝纤维化组比较,重度肝纤维化组患者血清中MALAT1 mRNA和CXCL14 mRNA表达水平升高(P<0.05),miR-150-5p表达水平降低(P<0.05);与对照组比较,TGF-β1组HSC中MALAT1 mRNA和CXCL14 mRNA表达水平均升高(P<0.05),miR-150-5p表达水平降低(P<0.05);与TGF-β1+si-NC组比较,TGF-β1+si-MALAT1组HSC中MALAT1mRNA和CXCL14 mRNA表达水平均降低(P<0.05),miR-150-5p表达水平升高(P<0.05);与TGF-β1+si-MALAT1组比较,TGF-β1+si-MALAT1+anti-miR-150-5p组HSC中miR-150-5p表达水平降低(P<0.05),CXCL14 mRNA表达水平升高(P<0.05);与TGF-β1+si-MALAT1组比较,TGF-β1+si-MALAT1+CXCL14组HSC中CXCL14 mRNA表达水平升高(P<0.05)。Western blotting法检测,与健康组比较,轻度和重度肝纤维化组患者血清中CXCL14蛋白表达水平升高(P<0.05);与轻度肝纤维化组比�Objective:To discuss the regulatory effect of long chain non-coding RNA(lncRNA)metastasis associated transcript 1(MALAT1)on the activation of the hepatic stellate cells(HSC)during the progression of liver fibrosis,and to clarify its mechanism.Methods:The serum samples were collected from 25 healthy volunteers(n=25,healthy group)and 25 liver fibrosis patients[mild liver fibrosis group(n=12)and severe liver fibrosis group(n=13)].The mouse HSC were divided into control group,transforming growth factor-β1(TGF-β1)group,TGF-β1+si-NC group,TGF-β1+si-MALAT1 group,TGF-β1+si-MALAT1+anti-miR-150-5p group,and TGF-β1 group+si-MALAT1+CXCL14 group.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of MALAT1 mRNA,microRNA-150-5p(miR-150-5p),and CXC chemokine ligand 14(CXCL14)mRNA in serum and HSC of the subjects in various groups;Western blotting method was used to detect the expression levels of CXCL14,α-smooth muscle actin(α-SMA),and typeⅠcollagenα1(COL1A1)proteins in serum of the subjects in various groups;CCK-8 method was used to detect the proliferation activities of the HSC in various groups;the expression levels ofα-SMA,COL1A1 proteins in the HSC in various groups were detected by immunofluorescence;the targeting relationship between miR-150-5p and MALAT1 and CXCL143′-UTR genes was detected by dual luciferase reporting system.Results.The RT-qPCR results showed that compared with healthy group,the serum expression levels of MALAT1 mRNA and CXCL14 mRNA of the subjects in mild and severe liver fibrosis groups were increased(P<0.05),while the expression levels of miR-150-5p were decreased(P<0.05);compared with mild liver fibrosis group,the serum expression levels of MALAT1 mRNA and CXCL14 mRNA of the subjects in severe liver fibrosis group were increased(P<0.05),while the expression level of miR-150-5p was decreased(P<0.05);compared with control group,the expression levels of MALAT1 mRNA and CXCL14 mRNA in the HSC in TGF-β1 group were increased(P<0.05),while the expr

关 键 词：肝纤维化 肝星状细胞 长链非编码RNA肺腺癌转移相关转录本1 微小RNA-150-5p CXC趋化因子配体14 

分 类 号：R575[医药卫生—消化系统]