MicroRNA-195/497对香烟烟雾提取物诱导的人支气管上皮细胞中TGF-β表达的影响  

Effects of microRNA-195/497 on the expression of TGF-βin human bronchial epithelial cells exposed to CSE

作　　者：迪拉拉·吐尔逊 祖力皮喀尔·阿卜杜热合曼 钟雪梅[1] 弓慧 茹克亚木·色麦提 李黎[1] Dilala Tuerxun;Zulipikaer Abudureheman;Zhong Xue-mei;Gong Hui;Rukeyamu Semaiti;Li Li(Department of Pulmonary and Critical Care Medicine,The First People's Hospital of Kashgar Prefecture,Kashgar,Xinjiang 844000,China;Xinjiang Clinical Research Center for Infectious Diseases(Tuberculosis),Kashgar,Xinjiang 844000,China)

机构地区：[1]喀什地区第一人民医院呼吸与危重症医学科,新疆喀什844000 [2]新疆感染性疾病(结核)临床医学研究中心,新疆喀什844000

出　　处：《中国现代医学杂志》2024年第14期1-8,共8页China Journal of Modern Medicine

基　　金：新疆维吾尔自治区自然科学基金(No:2021D01F12);喀什地区应用技术研究与开发计划(No:KS2021057)。

摘　　要：目的探讨microRNA-195/497(miR-195/497)在香烟烟雾提取物(CSE)诱导的人支气管上皮细胞中影响转化生长因子-β(TGF-β)表达的作用机制。方法用3%CSE诱导BEAS-2B细胞模拟慢性阻塞性肺疾病(COPD)的发病机制,实时荧光定量聚合酶链反应(qRT-PCR)检测miR-195、miR-497、SMAD7及TGF-β基因表达。双荧光素酶报告实验验证miR-195、miR-497与SMAD7的靶向关系,将miR-195mimic和miR-497 mimic转染至3%CSE处理的细胞中,qRT-PCR检测SMAD7的表达。将miR-195 mimic、miR-497 mimic和si-SMAD7转染至3%CSE处理的细胞中,qRT-PCR检测miR-195、miR-497、SMAD7、TGF-β的基因表达,Western blotting检测SMAD7、TGF-β的蛋白表达。结果3%CSE组miR-195、miR-497 mRNA相对表达量低于正常细胞组(P<0.05),SMAD7、TGF-βmRNA相对表达量高于正常细胞组(P<0.05)。miR-195 mimic组Wt-SMAD7 mRNA相对表达量低于mimic NC组(P<0.05)。miR-195 mimic组与mimic NC组Wt-SMAD7 mRNA相对表达量比较,差异无统计学意义(P>0.05)。miR-497 mimic组WtSMAD7 mRNA相对表达量低于mimic NC组(P<0.05)。miR-497 mimic组与mimic NC组Mut-SMAD7mRNA相对表达量比较,差异无统计学意义(P>0.05)。miR-195 mimic组SMAD7 mRNA相对表达量低于mimic NC组(P<0.05)。miR-497 mimic组SMAD7 mRNA相对表达量低于mimic NC组(P<0.05)。3%CSE组细胞miR-195和miR-497相对表达量较Control组下降(P<0.05),SMAD7及TGF-β蛋白相对表达量升高(P<0.05);miR-195 mimic+3%CSE组细胞miR-195相对表达量较3%CSE组升高(P<0.05),SMAD7、TGF-β下降(P<0.05),而miR-497相对表达量无明显差异(P>0.05);miR-497 mimic+3%CSE组细胞miR-497相对表达量较3%CSE组升高(P<0.05),SMAD7、TGF-β下降(P<0.05),而miR-195相对表达量无明显差异(P>0.05);si-SMAD7+3%CSE组SMAD7、TGF-β蛋白相对表达量较3%CSE组下降(P<0.05),而miR-195、miR-497无明显差异(P>0.05)。3%CSE组SMAD7、TGF-β蛋白相对表达量较Control组升高(P<0.05),miR-195 mimic+3%CSE组、miR-497 mimic+3%CSE组、si-SMAD7+3%CSE组较3%CSE组下�Objective To explore the mechanism underlying the effects of miR-195/497 on the expression of TGF-βin human bronchial epithelial cells exposed to cigarette smoke extract(CSE).Methods BEAS-2B cells were exposed to 3%CSE to simulate the pathogenesis of chronic obstructive pulmonary disease(COPD),and the expression levels of miR-195,miR-497,SMAD7 and TGF-βgenes were detected by quantitative real-time polymerase chain reaction(qRT-PCR).The targeting relationship of miR-195 and miR-497 with SMAD7 was validated via the dual luciferase reporter assay.After transfection of miR-195 mimic and miR-497 mimic into cells treated with 3%CSE,the expression of SMAD7 was detected by qRT-PCR.Following transfection of miR-195 mimic,miR-497 mimic and si-SMAD7 into cells treated with 3%CSE,the gene expressions of miR-195,miR-497,SMAD7 and TGF-βwere detected by qRT-PCR,and the protein expressions of SMAD7 and TGF-βwere measured via Western blotting.Results Compared with the normal cells,the relative expression levels of miR-195 and miR-497 were lower(P<0.01),and the relative expression levels of SMAD7 and TGF-βwere higher in cells treated with 3%CSE(P<0.05).The relative expression of Wt-SMAD7 in the miR-195 mimic group was lower than that in the mimic NC group(P<0.05).There was no difference in the relative expression of Mut-SMAD7 between the miR-195 mimic group and the mimic NC group(P>0.05).The relative expression of Wt-SMAD7 in the miR-497 mimic group was lower than that in the mimic NC group(P<0.05).There was no difference in the relative expression of Mut-SMAD7 between the miR-497 mimic group and the mimic NC group(P>0.05).The relative mRNA expression of SMAD7 in the miR-195 mimic group was lower than that in the mimic NC group(P<0.05).The mRNA relative expression of SMAD7 in the miR-497 mimic group was lower than that in the mimic NC group(P<0.05).The expressions of miR-195 and miR-497 were lower(P<0.05),and the expressions of SMAD7 and TGF-βwere higher in cells treated with 3%CSE than those in cells of the control group(P<

关 键 词：慢性阻塞性肺病 microRNA-195 microRNA-497 SMAD7 转化生长因子-Β 

分 类 号：R563.9[医药卫生—呼吸系统]