葛根素通过下调miR-23a促进小鼠前成骨细胞增殖和分化的研究  

Puerarin promotes the proliferation and differentiation of mouse osteoblasts by downregulating miR-23a

作　　者：蒋华[1] 李盼盼 张涛[1] 马良彧 邓烨 黎牧帆 马红兵[1] 罗婷[1] 颜宇[2] JIANG Hua;LI Pan-pan;ZHANG Tao;MA Liang-yu;DENG Ye;LI Mu-fan;MA Hong-bing;LUO Ting;YAN Yu(the Orthopedics Department of Chengdu Second People's Hospital,Chengdu 400016;the Department of Orthopedics,West China Hospital,Sichuan University,Chengdu 400016,China)

机构地区：[1]成都市第二人民医院骨科,四川成都400016 [2]四川大学华西医院骨科,四川成都400016

出　　处：《解剖科学进展》2023年第5期521-524,共4页Progress of Anatomical Sciences

基　　金：成都市卫健委课题(2020090)。

摘　　要：目的探讨葛根素是否通过下调miR-23a促进小鼠前成骨细胞增殖和分化。方法将小鼠前成骨细胞MC3T3-E1分为对照组、葛根素组、葛根素+miR-NC组和葛根素+miR-23a过表达组,RT-qPCR检测细胞中miR-23a的表达,CCK-8法检测细胞增殖活性,碱性磷酸酶活性检测葛根素对细胞活力的影响,Western blot检测细胞中Runx2蛋白的表达。应用生物信息学和双荧光素酶报告基因实验验证miR-23a和Runx2的靶向调控关系。结果与对照组相比,葛根素组MC3T3-E1细胞增殖活性增强,细胞中miR-23a表达降低,Runx2蛋白表达水平升高(P<0.05);与葛根素+miR-NC组相比,葛根素+miR-23a过表达组MC3T3-E1细胞增殖活性降低,细胞中Runx2蛋白表达降低(P<0.05);双荧光素酶报告基因实验证实miR-23a靶向负调控Runx2的表达。结论葛根素促进小鼠前成骨细胞增殖和分化,机制可能与下调miR-23a进一步调控Runx2表达有关。Objective To investigate whether puerarin promotes the proliferation and dfferentiation of mouse preosteoblasts by down-regulating miR-23a.Methods The mouse osteoblasts MC3T3-E1 was divided into control group,puerarin group,puerarin+miR-NC group and puerarin+miR-23a over expression group.The expression of miR-23a in cells was detected by RT-qPCR.The cell proliferation activity was detected by CCK8 method.The effect of puerarin on cell viability was detected by alkaline phosphatase activity,and the expression of Runx2 in cells was detected by Western blot.Bioinformatics and dual luciferase reporter gene experiments were used to verify the targeted regulatory relationship between miR-23a and Runx2.Results Compared with the control group,the proliferation activity of MC3T3-E1 cells in puerarin group was enhanced,the expression of miR-23a was decreased,and the expression of Runx2 was increased(P<0.05).Compared with puerarin+miR-NC group,the proliferation activity of MC3T3-E1 cells in the puerarin+miR-23a overexpression group was decreased,and the expression of Runx2 in the cells was decreased(P<0.05).Dual luciferase reporter gene assay confirmed the negative regulation of Runx2 expression by miR-23a targeting.Conclusion Puerarin promotes the proliferation and differentiation of mouse osteoblasts,and the mechanism may be related to down-regulation of miR-23a to further regulate the expression of Runx2.

关 键 词：葛根素 miR-23a 小鼠前成骨细胞 增殖 分化 

分 类 号：R336[医药卫生—人体生理学]