miR-181a-5p在皮肤成纤维样细胞衰老中的作用和机制  

The Role and Mechanism of miR-181a-5p in the Senescence of Skin Fibroblast-like Cells

作　　者：黄燕 杨艳清 万睿 周进飞 张层层 刘睿 许其军 张名 李慧娟 Huang Yan;Yang Yanqing;Wan Rui(Department of Plastic Surgery,Wuhan Third Hospital,Wuhan 430060,China)

机构地区：[1]武汉市第三医院整形外科,武汉430060 [2]襄阳市伊莱美医疗美容门诊部美容皮肤科,襄阳441099 [3]潜江市皮肤病防治院医学美容科,潜江433199 [4]武汉市第三医院皮肤科,武汉430060

出　　处：《华中科技大学学报（医学版）》2022年第6期830-834,889,共6页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：武汉市医学科研项目(No.WX20Q03)。

摘　　要：目的观察miR-181a-5p在H 2 O 2诱导大鼠皮肤成纤维样细胞衰老过程中的表达变化,并探讨其表达水平对细胞自噬和凋亡的影响。方法采用200μmol/L H_(2)O_(2)诱导大鼠皮肤成纤维样RS1细胞6 h,构建细胞衰老模型。将细胞分为5组:H 2 O 2处理组、mimic-NC组、mimic组、inhibitor-NC组和inhibitor组,采用脂质体转染技术将miR-181a-5p mimic、inhibitor等转染入相应的细胞中。MTT法检测细胞增殖活性;qPCR检测细胞miR-181a-5p表达水平。利用流式细胞术检测细胞凋亡;利用吖啶橙染色观察细胞自噬水平;采用Western blot检测细胞LC3B、p62、Beclin-1、自噬相关基因5(autophagy related 5,ATG5)蛋白表达水平。结果与正常RS1细胞比较,H 2 O 2诱导后的RS1细胞增殖活性显著降低(P<0.05),而miR-181a-5p表达水平显著升高(P<0.05)。与H 2 O 2处理组比较,mimic组RS1细胞排列紊乱,细胞轮廓不清晰,碎片较多,凋亡率显著升高(P<0.05),自噬水平降低,Beclin-1和ATG5表达显著降低(均P<0.05);inhibitor组RS1细胞轮廓清晰,过度伸展、空泡化的细胞减少,凋亡率显著降低(P<0.05),自噬水平升高,Beclin-1和ATG5表达显著升高(均P<0.05)。结论H 2 O 2诱导大鼠皮肤成纤维样细胞miR-181a-5p表达增加,且上调miR-181a-5p表达,可抑制细胞自噬、促进细胞凋亡。Objective To observe the expression of miR-181a-5p in rat skin fibroblast like cells induced by H_(2)O_(2),and to explore the effect of miR-181a-5p on autophagy and apoptosis.Methods Rat skin fibroblast-like RS1 cells were induced by 200μmol/L H_(2)O_(2) for 6 h to establish cell aging model.The cells were divided into 5 groups: H_(2)O_(2) treatment,mimic-NC,mimic,inhibitor-NC and inhibitor groups.miR-181a-5p mimic and inhibitor were transfected into the corresponding cells by liposome transfection technology.The transfection efficiency was detected by qPCR.MTT assay was used to detect cell proliferation.The expression level of miR-181a-5p was detected by qPCR.Apoptosis was detected by flow cytometry.Autophagy level was observed by acridine orange staining.The protein expression levels of LC3B,P62,Beclin-1 and autophagy related 5(ATG5)were detected by Western blotting.Results Compared with normal RS1 cells,the proliferation activity of RS1 cells induced by H_(2)O_(2) was significantly decreased(P<0.05),while the expression level of miR-181a-5p was significantly increased(P<0.05).Compared with the H_(2)O_(2) treatment group,RS1 cells in mimic group were disorderly arranged,with unclear cell outline and more fragments,the apoptosis rate was significantly increased(P<0.05),the autophagy level was decreased,the expression of Beclin-1 and ATG5 was significantly decreased(P<0.05);the outline of RS1 cells in inhibitor group was clear,the number of over extended and vacuolated cells was decreased,the apoptosis rate was significantly decreased(P<0.05),the autophagy level was increased,the expression of Beclin-1 and ATG5 was significantly increased(P<0.05).Conclusion After rat skin fibroblast-like cells are induced by H_(2)O_(2),the expression of miR-181a-5p was increased.And the up-regulated expression of miR-181a-5p can inhibit autophagy and promote apoptosis.

关 键 词：细胞衰老 H_(2)O_(2) miR-181a-5p 自噬 凋亡 

分 类 号：R751[医药卫生—皮肤病学与性病学]