摘要
目的探究miR-23b对气道平滑肌细胞(ASMCs)增殖的作用及其分子机制。方法组织块贴壁法分离培养BALB/c小鼠原代ASMCs,并进行传代。对ASMCs转染miR-23b mimic和inhibitor后,转化生长因子-β1(TGF-β1)以10μg/L的浓度干预处理各组转染细胞,评估miR-23b与TGF-β1对ASMCs增殖和凋亡的影响。采用生物信息学方法预测miR-23b靶基因,同时通过RT-PCR和Western blot分析检测miR-23b靶基因Smad3在ASMCs中的表达水平。结果 miR-23b的过表达能够显著抑制TGF-β1诱导的ASMCs增殖,促进细胞凋亡。RT-PCR和Western blot检测显示miR-23b负调控Smad3蛋白在ASMCs中的表达。结论 miR-23b可以通过Smad3信号通路抑制TGF-β1诱导的ASMCs增殖。
Objective To investigated the effects and mechanism of miR-23b on the proliferation of airway smooth muscle cells( ASMCs) proliferation.Methods Primary ASMCs from BALB/c mice were isolated and cultured by tissue-piece inoculation,and then passaged.After ASMCs were transfected with miR-23b mimics or miR-23b inhibitor,TGF-β1( 10 μg/L) were added to the transfected ASMCs,and the effects of miR-23b and TGF-β1 on ASMCs proliferation and apoptosis were evaluated through CCK8 and flow Cytometry,respectively.The target genes of miR-23b were predicted via bioinformatics software.Meanwhile,the expression of Smad3,which is the target gene of miR-23b,was determined using RT-PCR and Western blot.Results The overexpression of miR-23b significantly inhibited proliferation of ASMCs which induced by TGF-β1,and obviously promote the apoptosis.In addition,we also found that miR-23b negatively regulated the expression of Smad3.Conclusion miR-23b could suppress the proliferation of ASMCs induced by TGF-β1 via Smad3 signaling pathway.
作者
张炜
黄林洁
陈茗
ZHANG Wei;HUANG Lin-jie;CHEN ming(Department of Respiratory,Xili People's Hospital,Nanshan District,Shenzhen 518055,Guangdong,China)
出处
《广东医学》
CAS
2019年第1期80-86,共7页
Guangdong Medical Journal
基金
国家自然科学基金资助项目(编号:81500018)
广东省自然科学基金资助项目(编号:2016A030313332)
