miR-218-5p通过靶向下调ANKRD1抑制地塞米松诱导的人成骨细胞HFOB1.19损伤  

miR-218-5p inhibits injury of dexamethasone-induced human osteoblasts HFOB1.19 by down-regulating ANKRD1

作　　者：陈晨[1] 郑润泉 李宗玉[1] CHEN Chen;ZHENG Runquan;LI Zongyu(Department of Orthopedics,960th Hospital of PLA Joint Logistics Support Force,Jinan 250031,China)

机构地区：[1]中国人民解放军联勤保障部队第960医院骨科,济南250031

出　　处：《中国免疫学杂志》2022年第19期2325-2329,2335,共6页Chinese Journal of Immunology

摘　　要：目的:探讨miR-218-5p对地塞米松诱导的人成骨细胞损伤的影响和可能机制。方法:体外培养人成骨细胞HFOB1.19,分别转染miR-218-5p模拟物、ANKRD1小干扰RNA或共转染miR-218-5p模拟物与ANKRD1过表达载体,然后采用10μmol/L地塞米松干预转染后的细胞24 h,RT-qPCR检测细胞中miR-218-5p和ANKRD1 mRNA表达水平,流式细胞术检测细胞凋亡,Western blot检测细胞中Bcl-2和Bax蛋白表达水平,ELISA检测细胞培养上清中IL-6、IL-1β和TNF-α水平。双荧光素酶报告基因实验验证ANKRD1和miR-218-5p的调控关系。结果:地塞米松抑制成骨细胞中miR-218-5p表达,促进ANKRD1表达(P<0.05)。过表达miR-218-5p或敲减ANKRD1降低了地塞米松诱导的成骨细胞凋亡率及Bax蛋白、IL-6、IL-1β和TNF-α表达(P<0.05),促进了Bcl-2蛋白表达(P<0.05)。过表达ANKRD1逆转了过表达miR-218-5p对地塞米松诱导的成骨细胞凋亡及炎症因子IL-6、IL-1β和TNF-α表达的抑制作用。双荧光素酶报告基因实验结果显示,miR-218-5p可靶向结合ANKRD1;过表达miR-218-5p抑制了成骨细胞凋中ANKRD1表达。结论:miR-218-5p可通过靶向抑制ANKRD1减少地塞米松诱导的成骨细胞凋亡及炎症因子表达,减轻成骨细胞损伤。Objective:To investigate the effect and possible mechanism of miR-218-5p on injury of dexamethasone-induced human osteoblasts.Methods:Human osteoblasts HFOB1.19 were cultured in vitro and transfected with miR-218-5p mimic,ANKRD1small interfering RNA or co-transfected with miR-218-5p mimic and ANKRD1 overexpression vector,and then used 10μmol/L dexamethasone interfered with transfected cells for 24 hours.Expression levels of miR-218-5p and ANKRD1 mRNA in cells were detected by RT-qPCR.Cells apoptosis was detected by flow cytometry.Protein expressions of Bcl-2 and Bax in cells were detected by Western blot.Expression levels of IL-6,IL-1βand TNF-αin cell culture supernatant were detected by ELISA.Dual luciferase reporter gene experiment verified the regulatory relationship between ANKRD1 and miR-218-5p.Results:Dexamethasone inhibited expression of miR-218-5p in osteoblasts(P<0.05),while promoted expression of ANKRD1(P<0.05).Overexpression of miR-218-5p or knockdown of ANKRD1 reduced apoptosis rate of dexamethasone-induced osteoblasts and expressions of Bax protein,IL-6,IL-1βand TNF-α(P<0.05),while promoted expression of Bcl-2 protein(P<0.05).Overexpression of ANKRD1 reversed the inhibitory effect of overexpression of miR-218-5p on dexamethasone-induced osteoblast apoptosis and expressions of inflammatory factors IL-6,IL-1βand TNF-α.Results of dual luciferase reporter gene experiments showed that miR-218-5p could target ANKRD1;overexpression of miR-218-5p inhibited expression of ANKRD1 in osteoblasts.Conclusion:miR-218-5p can inhibit dexamethasone-induced osteoblast apoptosis and expressions of inflammatory factors by down regulating ANKRD1,which reduces osteoblast injury.

关 键 词：人成骨细胞 miR-218-5p ANKRD1 细胞凋亡 炎症 

分 类 号：R681[医药卫生—骨科学]