miR-100-5p通过抑制大鼠骨髓间充质干细胞的迁移和成骨分化参与非创伤性股骨头坏死  

miR-100-5p is involved in non-traumatic osteonecrosis of the femoral head by inhibiting the migration and osteogenic differentiation of BMSCs

作　　者：杨武 杨韵霏 郭民康 朱卫文[1] 赵安平 白浩波 张健[1] YANG Wu;YANG Yunfei;GUO Minkang;ZHU Weiwen;ZHAO Anping;BAI Haobo;ZHANG Jian(Department of Orthopedics,the First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学附属第一医院骨科,重庆400016

出　　处：《细胞与分子免疫学杂志》2022年第2期159-164,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：重庆市技术创新与应用发展专项(cstc2019jscx-msxmX2045)。

摘　　要：目的探讨miR-100-5p在非创伤性股骨头坏死(NONFH)发病机制中的作用。方法实时定量PCR检测NONFH患者的miRNA表达,确定NONFH患者股骨头组织miR-100-5p高表达。而后培养大鼠骨髓间充质干细胞(rBMSC)分为空白对照组,地塞米松处理组(地塞米松处理3 d),微小RNA阴性对照组(转染miR-NC),agomiR-100-5p组(miR-100-5p过表达),antagomiR-100-5p组(转染miR-100-5p拮抗剂)。使用实时定量PCR检测股骨头骨组织miR-100-5p、碱性磷酸酶(ALP)、Runt相关转录因子2(RUNX2)和Ⅰ型胶原蛋白(Col1)的mRNA表达。采用Western blot法检测股骨头骨组织ALP、RUNX2、Col1和骨形态发生蛋白受体2(BMPR2)蛋白的表达。采用划痕愈合实验评估miR-100-5p对rBMSC迁移能力的影响。通过茜素红染色观察miR-100-5p对rBMSC成骨分化能力的影响。结果与正常的股骨头骨组织相比,NONFH患者股骨头骨组织中miR-100-5p显著上调;与正常rBMSC相比,20μmol/L地塞米松处理后的rBMSC中miR-100-5p表达上调;rBMSC中miR-100-5p的上调降低ALP、RUNX2、Col1和BMPR2的表达,抑制rBMSC的成骨分化能力和迁移能力。结论miR-100-5p在NONFH患者的骨组织和20μmol/L地塞米松处理后的rBMSC中表达升高。上调的miR-100-5p可能通过抑制rBMSC的迁移和成骨分化参与NONFH的发病机制。Objective To investigate the role of miR-100-5p in the pathogenesis of non-traumatic osteonecrosis of the femoral head(NONFH). Methods The miRNA expression in patients with NONFH was detected by real-time quantitative PCR, the high expression of miR-100-5p in femoral head tissues of the patients determined. Rat bone marrow mesenchymal stem cells(rBMSCs) were cultured and divided into 5 groups: blank control group, dexamethasone treatment group(treated with dexamethasone for 3 days), miR-NC group(transfected with miR-NC), agomiR-100-5p group(overexpression of miR-100-5p), and antagomiR-100-5p group(transfected with miR-100-5p antagonist). The mRNA expression levels of miR-100-5p, alkaline phosphatase(ALP), Runt-associated transcription factor 2(RUNX2), and collagen type Ⅰ(Col1) were detected by real-time quantitative PCR. The protein expressions of ALP, RUNX2, Col1, and bone morphogenetic protein receptor 2(BMPR2) were detected by Western blotting. The effect of miR-100-5p on the migration ability of rBMSCs was evaluated by scratch healing assay. And the effect of miR-100-5p on osteogenic differentiation ability of rBMSCs was investigated by alizarin red staining. Results miR-100-5p was significantly upregulated in the femoral head bone tissue of NONFH patients compared with normal femoral head bone tissue. Compared with those in the normal rBMSCs, the expression of miR-100-5p in rBMSCs treated with 20 μmol/L of dexamethasone was up-regulated. The upregulation of miR-100-5p in rBMSCs reduced the expressions of ALP, RUNX2, Col1, and BMPR2, and inhibited the osteogenic differentiation and migration abilities of rBMSCs. Conclusion The expression of miR-100-5p is elevated in bone tissues of NONFH patients and in rBMSCs treatedwith 20 μmol/L of dexamethasone. The up-regulated miR-100-5p may be involved in the pathogenesis of NONFH by inhibiting the migration and osteogenic differentiation of rBMSCs.

关 键 词：大鼠骨髓间充质干细胞(rBMSC) 非创伤性股骨头坏死(NONFH) miR-100-5p 细胞迁移 成骨分化 

分 类 号：R392.12[医药卫生—免疫学] Q255[医药卫生—基础医学]