长链非编码RNA H19抑制地塞米松致成骨细胞凋亡的作用  

Effects of long non-coding RNA H19 on apoptosis of osteoblasts induced by dexamethasone

作　　者：杨慧霞 白志刚 迟宏扬[1,2] 马天龙 马胜超 姜怡邓 Yang Huixia;Bai Zhigang;Chi Hongyang;Ma Tianlong;Ma Shengchao;Jiang Yideng(Clinical School of Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;National Health Commission Key Laboratory of Metabolic Cardiovascular Disease Research,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Peoples Hospital of Ningxia Hui Autonomous Region,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Basic School of Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China)

机构地区：[1]宁夏医科大学临床医学院,宁夏回族自治区银川市750004 [2]国家卫生健康委代谢性心血管疾病研究重点实验室,宁夏回族自治区银川市750004 [3]宁夏回族自治区人民医院,宁夏回族自治区银川市750004 [4]宁夏医科大学基础医学院,宁夏回族自治区银川市750004

出　　处：《中国组织工程研究》2023年第28期4513-4518,共6页Chinese Journal of Tissue Engineering Research

基　　金：宁夏回族自治区重点研发计划项目(2020BFH02001),项目负责人:白志刚;国家自然科学基金(82060412),项目负责人:白志刚;中国医学科学院中央级公益性科研院所基本科研业务费项目(2019PT330002),项目负责人:姜怡邓。

摘　　要：背景:激素性股骨头坏死的发病机制尚不清楚,成骨细胞凋亡与其关系密切。目的:探讨长链非编码RNA H19(long Non-Coding RNA H19,lncRNA H19)在地塞米松促进成骨细胞凋亡中的作用。方法:培养小鼠成骨细胞MC3T3-E1,分为对照组(不处理)和地塞米松组(1μmol/L地塞米松处理24 h),采用Western blot检测各组细胞Bax、Bcl-2和细胞外调节蛋白激酶(p-ERK1/2/ERK1/2)蛋白的表达,流式细胞术和细胞活力染色检测细胞凋亡情况,qRT-PCR检测lncRNAH19的表达。转染lncRNA H19阴性对照(ad-NC)和lncRNA H19过表达质粒(ad-lncRNA H19)并给予地塞米松干预24 h后,采用细胞活力染色、Western blot和流式细胞术检测MC3T3-E1细胞凋亡情况;使用MAPK-ERK通路抑制剂PD98059处理MC3T3-E1细胞24 h,Western blot检测各组细胞Bax和Bcl-2蛋白的表达。结果与结论:(1)与对照组相比,地塞米松组MC3T3-E1细胞凋亡率升高,lncRNA H19相对表达降低(P<0.01),MAPK/ERK信号通路被抑制;(2)上调MC3T3-E1细胞lncRNA H19后,细胞凋亡率降低(P<0.01),MAPK/ERK信号通路被激活;(3)与地塞米松+ad-lncRNA H19组相比,地塞米松+ad-lncRNA H19+PD98059组细胞Bax蛋白表达增加(P<0.01),Bcl-2蛋白表达降低(P<0.01);(4)提示过表达lncRNA H19通过激活MAPK-ERK信号通路抑制地塞米松诱导的MC3T3-E1细胞凋亡。BACKGROUND:The pathogenesis of steroid-induced osteonecrosis of femoral head is still unclear,which is closely related to osteoblast apoptosis.OBJECTIVE:To investigate the role of long non-coding RNA H19(lncRNA H19)in dexamethasone-induced osteoblast apoptosis.METHODS:Mouse osteoblasts(MC3T3-E1)were cultured and divided into control group(no treatment)and dexamethasone group(treated with 1μmol/L dexamethasone for 24 hours).Western blot was used to detect the protein expression levels of Bax,Bcl-2,and p-ERK1/2/ERK1/2.Flow cytometry and cell viability staining were used to detected cell apoptosis.qRT-PCR was performed to detect the expression of lncRNA H19.lncRNA H19 negative control plasmid(ad-NC)and lncRNA H19 overexpression plasmid(ad-lncRNA H19)were transfected into MC3T3-E1 cells,followed by 24 hours of dexamethasone intervention.Cell apoptosis of MC3T3-E1 was detected using cell viability staining,western blot assay,and flow cytometry.MC3T3-E1 cells were treated with MAPK-ERK pathway inhibitor(PD98059)for 24 hours and western blot was used to detect the protein expression levels of Bax and Bcl-2.RESULTS AND CONCLUSION:Compared with the control group,the apoptotic rate of MC3T3-E1 cells was increased in the dexamethasone group(P<0.01),while the expression of lncRNA H19 was decreased(P<0.01)and the MAPK/ERK signaling pathway was inhibited.U p-regulation of lncRNA H19 in MC3T3-E1cells reduced the apoptosis of cells(P<0.01)and activated the MAPK/ERK signaling pathway.MC3T3-E1 cells treated with dexamethason e+ad-lncRNA H19+PD98059 showed an increase in the expression of Bax(P<0.01)and a decrease in the expression of Bcl-2(P<0.01)compared with the cells treated with dexamethasone+ad-lncRNA H19.These findings indicate that overexpression of lncRNA H19 can inhibit dexamethasone-induced apoptosis in MC3T3-E1 cells by activating the MAPK-ERK signaling pathway.

关 键 词：lncRNA H19 MAPK/ERK信号通路 地塞米松 MC3T3-E1细胞 凋亡 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R681