摘要
目的探讨miRNAs和CTNNB1在激素性股骨头坏死患者中的表达水平,以及miR-124-3p对骨髓间充质干细胞成骨分化的影响。方法选择2020年9月至2022年5月就诊于新疆医科大学第五附属医院行髋关节置换术的患者30例,激素性股骨头坏死患者15例作为实验组,股骨颈骨折患者15例作为对照组。术中扩髓时收集患者的骨髓组织,提取总mRNA,应用qRT-PCR检测各组miR-322-5p、miR-124-3p、miR-125a-3p及CTNNB1的表达量,用Western Blot检测CTNNB1蛋白的表达量。在骨髓间充质干细胞(BMSCs)中转染miR-124-3p的模拟物和抑制剂来检测其对BMSCs分化的影响作用,并验证miR-124-3p与CTNNB1的靶标关系。结果在实验组BMSCs中miR-125a-3p、CTNNB1表达量高于对照组(P<0.05);miR-124-3p的表达量低于对照组(P<0.001),miR-322-5p的表达量在两组中无明显差异。转染miR-124-3p+mimics组的miR-124-3p表达量高于其他组,CTNNB1表达量低于其他组(P<0.001),转染miR-124-3p inhibitor组的结果则相反。分化能力评估发现过表达miR-124-3p可以促进BMSCs成骨分化能力,抑制表达则结果相反。双荧光素酶报告系统结果显示,miR-124-3p与CTNNB1有靶向结合关系。结论miR-124-3p可以靶向CTNNB1,并且miR-124-3p可以调控骨髓间充质干细胞并促进其成骨分化。
Objective To investigate the expression levels of miRNAs and CTNNB1 in patients with steroid-induced necrosis of the femoral head,and the effect of miR-124-3p on osteogenic differentiation of bone marrow mesenchymal stem cells.Methods Thirty patients who underwent total hip replacement,15 patients with steroid-induced necrosis of the femoral head were selected as the experimental group and 15 patients with femoral neck fracture were selected as the control group from September 2020 to May 2022 in the Fifth Affiliated Hospital of Xinjiang Medical University.Bone marrow tissues of patients were collected during intraoperative bone marrow aspiration and total mRNA were extracted.qRT-PCR was used to detect the expression levels of miR-322-5p,miR-124-3p,miR-125a-3p and CTNNB1 in each group,and Western Blot was used to detect the expression levels of CTNNB1.Mimics and inhibitors of miR-124-3p were transfected into bone marrow mesenchymal stem cells(BMSCs)to detect its effect on BMSCs differentiation and verify the relationship between miR-124-3p and CTNNB1 targets.Results The expression levels of miR-125a-3p and CTNNB1 in BMSCs in the experimental group were higher than those in the control group(P<0.05).The expression level of miR-124-3p was lower than that of the control group(P<0.001),and the expression level of miR-322-5p was not significantly different between the two groups.The expression of miR-124-3p and CTNNB1 in the miR-124-3p+mimics group was higher than that in other groups(P<0.001),but the results of the miR-124-3p inhibitor group were opposite.Differentiation ability evaluation showed that overexpression of the miR-124-3p could promote the osteogenic differentiation ability of BMSCs,while inhibition of expression showed the opposite result.The results of the dual luciferase reporting system showed that miR-124-3p had a targeted binding relationship with CTNNB1.Conclusion miR-124-3p can target CTNNB1,and miR-124-3p can regulate bone marrow mesenchymal stem cells and promote osteogenic differentiation.
作者
杨增强
郝飞虎
原天祎
周治衡
崔泳
Yang Zengqiang;Hao Feihu;Yuan Tianyi;Zhou Zhiheng;Cui Yong(Department of Orthopaedic Center,The Fifth Affiliated Hospital of Xinjiang Medical University,Urumqi Xinjiang,830011,China)
出处
《生物骨科材料与临床研究》
CAS
2024年第1期7-13,共7页
Orthopaedic Biomechanics Materials and Clinical Study
基金
国家自然科学基金(81960396)
新疆医科大学研究生创新项目(CXCY2022014)。
