摘要
目的:探究环状核糖核酸(RNA)zeste基因增强子同源物2(circEZH2)调控骨形态发生蛋白2(BMP2)对骨髓间充质干细胞(BMSCs)成骨分化的影响。方法:使用生长培养基培养骨髓间充质干细胞,将骨髓间充质干细胞随机分为对照组、si-NC组、si-circEZH2组、si-circEZH2+si-NC组和si-circEZH2+si-BMP2组;qRT-PCR法检测各组骨髓间充质干细胞中circEZH2及BMP2 mRNA表达水平,CCK-8法检测各组骨髓间充质干细胞增殖能力,流式细胞仪检测各组骨髓间充质干细胞凋亡情况,碱性磷酸酶(ALP)活性检测试剂盒检测ALP活性,蛋白免疫印迹法(Western Blotting)检测各组骨髓间充质干细胞中BMP2、成骨相关蛋白骨钙素(OC)、RUNT相关转录因子2(Runx2)和骨桥蛋白(OPN)表达水平。结果:相较于对照组和si-NC组,si-circEZH2组骨髓间充质干细胞中circEZH2的表达水平、细胞增殖抑制率、凋亡率显著降低(P<0.05),BMP2 mRNA表达水平、ALP活性、BMP2、OC、Runx2及OPN表达显著升高(P<0.05);相较于si-circEZH2组,si-circEZH2+si-NC组及si-circEZH2+si-BMP2组骨髓间充质干细胞中circEZH2 mRNA的表达水平、细胞增殖抑制率、凋亡率显著升高(P<0.05),BMP2 mRNA表达水平、ALP活性、BMP2、OC、Runx2、OPN表达显著降低(P<0.05)。结论:circEZH2低表达可能通过促进BMP2过表达来增强骨髓间充质干细胞成骨分化,从而改善骨质疏松症。
Objective:To explore the efficacy of circular RNA zeste gene enhancer homolog 2(circEZH2)regulating bone morphogenetic protein 2(BMP2)on the osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods:BMSCs cells were cultured with growth medium,and BMSCs cells were randomly divided into control group,si-NC group,si-circEZH2 group,si-circEZH2+si-NC group and si-circEZH2+si-BMP2 group.qRT-PCR method was used to detect the expression levels of circEZH2 and BMP2 mRNA in BMSCs cells in each group,CCK-8 method was used to detect cell proliferation ability of BMSCs in each group,flow cytometry was used to detect the apoptosis of BMSCs in each group,alkaline phosphatase(ALP)activity detection kit was used to detect ALP activity,Western Blotting was used to detect the expression levels of BMP2,osteogenic proteins osteocalcin(OC),RUNT related transcription factor2(Runx2)and osteopontin(OPN)in BMSCs cells of each group.Results:Compared with the control group and the si-NC group,the expression level of circEZH2,the cell proliferation inhibition rate and the apoptosis rate in the BMSCs of the si-circEZH2 group were significantly reduced(P<0.05),and the expression level of BMP2 mRNA,ALP activity,the expression of BMP2,OC,Runx2,OPN proteins were significantly increased(P<0.05).Compared with si-circEZH2 group and si-circEZH2+si-NC group,the expression level of circEZH2,the cell proliferation inhibition rate and the apoptosis rate in the BMSCs of the si-circEZH2+si-BMP2 group were significantly increased(P<0.05),and the expression level of BMP2 mRNA,ALP activity,the expression of BMP2,OC,Runx2,OPN proteins were significantly reduced(P<0.05).Conclusion:The low expression of circEZH2 may enhance the osteogenic differentiation of BMSCs by promoting the overexpression of BMP2 to improve osteoporosis.
作者
阮锋
李贺伟
龚艳琳
刘佳丽
刘平
RUAN Feng;LI Hewei;GONG Yanlin;LIU Jiali;LIU Ping(Department of Osteology,Liyuan Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430077,China;Department of Endocrinology,Wuhan First Hospital,Wuhan 430030,China)
出处
《中国中医骨伤科杂志》
CAS
2022年第10期7-12,共6页
Chinese Journal of Traditional Medical Traumatology & Orthopedics
基金
武汉市卫生健康委员会资助项目(武卫通[2019]87号)。
关键词
骨质疏松
环状RNA
zeste基因增强子同源物2
骨形态发生蛋白2
骨髓间充质干细胞
成骨分化
增殖
凋亡
碱性磷酸酶
osteoporosis
circular RNA zeste gene enhancer homolog 2
bone morphogenetic protein 2
bone marrow mesenchymal stem cells
osteogenic differentiation
proliferation
apoptosis
alkaline phosphatase(ALP)
