miR-186-5p通过抑制Twist1诱导人皮肤成纤维细胞衰老  

miR-186-5p Induces Senescence in Human Skin Fibroblasts by Inhibiting Twist1

作　　者：彭亚婷 熊一峰[2] 王淑华 刘志刚[1] 张志彬 张颖鹏[1] PENG Yating;XIONG Yifeng;WANG Shuhua;LIU Zhigang;ZHANG Zhibing;ZHANG Ying peng(Department of Dermatology,the Second Affiliated Hospital of Nanchang University,Nanchang 330006,China;Department of Pathology,the First Affiliated Hospital of Nanchang University,Nanchang 330006,China)

机构地区：[1]南昌大学第二附属医院皮肤科,江西南昌330006 [2]南昌大学第一附属医院病理科,江西南昌330006

出　　处：《中国皮肤性病学杂志》2021年第1期7-14,共8页The Chinese Journal of Dermatovenereology

基　　金：国家自然科学基金(81860548);江西省教育厅科学基金(GJJ170146)。

摘　　要：目的探讨miR-186-5p对人皮肤成纤维细胞衰老的影响及其机制。方法采用实时定量PCR方法检测miR-186-5p的表达,应用衰老相关β-半乳糖苷酶(SA-β-gal)染色、CCK-8和EdU等方法研究miR-186-5p对人真皮成纤维细胞(HDFs)衰老的作用。通过生物信息学分析、荧光素酶报告试验、实时定量PCR、Western blot和免疫组化法证实miR-186-5p的下游靶基因。结果首次发现miR-186-5p在传代衰老的HDFs和老年人皮肤真皮组织中表达上调。miR-186-5p的过表达抑制了HDFs细胞的增殖,同时增强了衰老相关指标p16、p21和SA-β-gal的表达。双荧光素酶报告分析显示Twist1是miR-186-5p的下游靶基因。结论miR-186-5p可通过抑制Twist1诱导HDFs衰老,其可能成为干预皮肤老化的潜在靶点。Objective To investigate the role of miR-186-5p on the aging of human dermal fibroblasts and its mechanism.Methods The expression of miR-186-5p was measured by quantitative real-time PCR(qRT-PCR).Senescence-associatedβ-galactosidase(SA-β-gal)staining,the cell counting Kit-8(CCK-8)and 5-ethynyl-2'-deoxyuridine(EdU)were used to investigate the effects of miR-186-5p on human dermal fibroblast(HDF)senescence.The target gene of miR-186-5p was confirmed by bioinformatics analysis,luciferase reporter assay,qRT-PCR,Western blot and immunohistochemical analysis.Results We first found that miR-186-5p was up-regulated in passage-aged HDFs(pHDFs)and skin dermal tissues from old donors(old skin).Overexpression of miR-186-5p inhibited HDFs proliferation while enhanced the expression of p16,p21 and the SA-β-gal-positive ratio.Dual-luciferase reporter assays showed that Twist1 was the downstream target gene of miR-186-5p.Conclusion miR-186-5p could induce HDF senescence by inhibiting Twist1.It could be a potential therapeutic target for preventing skin aging.

关 键 词：miRNA-186 MICRORNA TWIST 衰老 皮肤 

分 类 号：R751[医药卫生—皮肤病学与性病学]