miR-21-5p在H_(2)O_(2)诱导的MC3T3-E1成骨细胞凋亡中的作用  

Role of miR-21-5p on H_(2)0_(2)-induced Apoptosis of MC3T3-E1 Osteoblasts

作　　者：戴峰 王青[2] 王江 范志豪 蒋正宇 DAI Feng;WANG Qing;WANG Jiang;FAN Zhihao;JIANG Zhengyu(Department of Orthopedics,Liyang Branch,Jiangsu Province People's Hospital,Liyang,Jiangshu,213300,China;Department of Joint Surgery,Jiangsu Province People’s Hospital,Nanjing,210029,China)

机构地区：[1]江苏省人民医院溧阳分院骨科,江苏省溧阳市213300 [2]江苏省人民医院关节外科,南京市210029

出　　处：《医学分子生物学杂志》2020年第6期416-421,共6页Journal of Medical Molecular Biology

基　　金：江苏省卫生厅科研基金(No.2016D1472)。

摘　　要：目的探讨microRNA(miR)-21-5p通过核因子E2相关因子2(Nrf2)对H202诱导的MC3T3-E1成骨细胞凋亡的影响。方法实验分为4组:对照组、H_(2)0_(2) 组、H_(2)0_(2) +mimic组和 H_(2)0_(2) +inhibitor组。MTT检测细胞活力,Hoechst染色和流式细胞术检测细胞凋亡,ELISA检测SOD、LDH、MDA的含量,Western蛋内印迹检测Bd-2、Bax、Osx、Col1 A1、Nrf2、p-Nrf2和HO-1。结果与对照组比较,H202组细胞活力显著降低(P<0.05);凋亡显著增加(p<0.05);氧化应激显著增加(P<0.05);成骨细胞特异性转录因子Osx、Col1 A 1和氧化应激通路蛋白Nrf2、P-Nrf2和H0-1显著下调(P<0.05)。与H202组比较,H202+mimic逆转上述结果,而加入inhibitor逆转了H202+mimic组的结果(P<0.05)。结论miR-21-5p通过活化N rf2抑制H202诱导M C3T3-E1成骨细胞凋亡。Objective To investigate the effect of MicroRNA(miR)-21-5p on H202-induced apoptosis of MC3T3-E1 osteoblasts through nuclear factor erythroid 2-related factor 2(Nrf2).Methods The cells were divided into control group,H202 group,H202+mimic group and H202+inhibitor group.Cell viability was detected by MTT.Cell apoptosis was detected by Hoechst staining and flow cytometry.SOD,LDH and MDA contents were detected by ELISA.Bcl-2,B ax,Osx,Col1 A1,Nrf2,P-NRf2 and Ho-1 were detected by Western blotting.Results Compared with the control group,the cell activity of H202 group was significantly decreased(P<0.05);apoptosis was significantly increased(P<0.05);oxidative stress was significantly increased(P<0.05);osteoblasts specific transcription factors Osx,Coll A 1 and oxidative stress pathway proteins Nrf2,p-Nrf2 and HO-1 were significantly down-regulated(P<0.05).H202+mimic reversed the effects of H202,which was reversed with miRNA inhibitor(P<0.05).Conclusion miR-21-5p inhibits apoptosis of H202-induced MC3T3-E1 osteoblasts by activating Nrf2 pathway.

关 键 词：miR-21-5p Nrf2通路 成骨细胞 凋亡 

分 类 号：R363.21[医药卫生—病理学]