miR-330-5p负性调控ADAM17基因抑制人心脏微血管内皮细胞的间质转化  

MiR-330-5p down-regulated ADAM17 expression in human cardiac microvascular endothelial cells and inhibited endothelial-to-mesenchymal transition

作　　者：鞠延玲[1] 朱国伟[1] 赵旭[1] 臧雪莲[1] 

机构地区：[1]锦州市中心医院,辽宁锦州121001

出　　处：《贵州医药》2016年第10期1011-1014,共4页Guizhou Medical Journal

摘　　要：目的探讨人心脏微血管内皮细胞(HCMEC)间质转化(EndMT)过程中miR-330-5p和ADAM17基因的表达,阐明miR-330-5p对EndMT的影响。方法培养HCMEC,应用TGF-β1诱导后,利用免疫荧光化学检测CD31和α-平滑肌肌动蛋白(α-SMA)在细胞中的表达。运用生物信息学方法对miR-330-5p和ADAM17基因的配对关系进行预测。脂质体2000转染miR-330-5p模拟物后,Real-time PCR检测miR-330-5p、ADAM17和α-SMA mRNA的表达,Western blotting检测ADAM17和α-SMA蛋白的表达。结果光镜下可见诱导后细胞由鹅卵石形态变为长梭形,免疫荧光化学显示诱导前有强烈的CD31表达,诱导后出现α-SMA的高表达。生物信息学软件TargetScan和miRanYda显示miR-330-5p和ADAM17基因二者靶向配对良好。Real-time PCR和Western blotting检测结果均表明过表达miR-330-5p能够降低ADAM17和α-SMA mRNA及蛋白的表达。结论 miR-330-5p通过下调人心脏微血管内皮细胞中ADAM17基因的表达,进而抑制TGF-β1诱导的EndMT。Objective To identify expressions of miR-330-5p and ADAM17 in human cardiac microvascular endothelial cells（HCMEC）and explore the role of miR-330-5p on endothelial-to-mesenchymal transition（EndMT）which may regulate ADAM17 expression.Methods HCMEC were purchased and then cultured in vitro.Cell morphology was observed with phase contrast microscope and expressions of CD31andα-SMA in HCMEC were determined by immunofluorescence cytochemistry.MiR-330-5p which may regulate ADAM17 expression was predicted by bioinformatics.After transfection of miR-330-5p mimics into cells,the expressions of miR-330-5p,ADAM17andα-SMA were determined by real-time PCR and western blotting（WB）.Results Before the induction of TGF-β1in HCMEC,CD31 expression was positive.However,α-SMA was found after the induction by immunofluorescence cytochemistry.Real-time PCR and western blotting was showed over-expression of miR-330-5p down-regulated ADAM17andα-SMA expressions.Conclusion MiR-330-5p down-regulated ADAM17 expression in HCMEC and then inhibit EndMT.

关 键 词：微小RNA 解整合素-金属蛋白酶17 内皮间质转化 人心脏微血管内皮细胞 

分 类 号：R331.3[医药卫生—人体生理学]