安徽医科大学学报2025，Vol.60Issue(6)：1059-1068,10.DOI:10.19405/j.cnki.issn1000-1492.2025.06.012
lncRNA H19调控miR-149-5p/BMP2轴对BMSCs的成骨分化及足骨骨折小鼠骨再生的影响
The role of lncRNA H19 in regulating the miR-149-5p/BMP2 axis on osteogenic differentiation of BMSCs and bone regeneration in metatarsal fracture
摘要
Abstract
Objective To investigate the role and mechanism of long non-coding RNA(lncRNA)H19 in osteogen-ic differentiation of bone marrow mesenchymal stem cells(BMSCs)and bone regeneration in mice with foot frac-ture.Methods BMSCs were divided into microrNA-149-5p(miR-149-5p)-mimic negative control(miR-mimic-NC)group and miR-149-5p-mimic(miR-mimic)group and transfected with lncRNA H19-wt and lncRNA H19,respectively.Mut recombinant plasmid,or bone morphogenetic protein 2(BMP2)3'UTR-wt and BMP2 3'UTR-mut recombinant plasmid were transfected,respectively.Dual-luciferase reporter gene assay was used to detect the luciferase activity of each group.To study the regulatory effect and mechanism of lncRNA H19 on osteogenic differ-entiation in vitro,BMSCs were divided into control group and osteogenic induction group(osteogenic group).After osteogenic induction,BMSCs were transfected with corresponding plasmids and divided into osteogenic+si-NC group,osteogenic+si-H19 group,osteogenic+si-H19+miR-inhibitor-NC group,osteogenic+si-H19+miR-in-hibitor group,osteogenic+si-H19+pcDNA-NC group,and osteogenic group si-H19+pcDNA-BMP2 group.Os-teogenic differentiation was evaluated by alkaline phosphatase(ALP)activity assay and alizarin red staining.A mouse foot fracture model was established,and 36 mice were randomly divided into sham operation group,model group,model+pcD-null group,and model+pcD-H19 group,with 9 mice in each group.Osteogenic differentia-tion was assessed by ALP activity assay.Real-time fluorescent quantitative PCR(qPCR)was used to detect the ex-pression of lncRNA H19,miR-149-5p,and BMP2.Western blot was used to detect the expression of BMP2,OCN,OSX,RUNX2,and OPN.Results In cells transfected with lncRNA H19-wt,the luciferase activity in miR-mimic group was lower than that in miR-mimic-NC group(P＜0.05).The luciferase activity of miR-mimic group was lower than that of miR-mimic-NC group(P＜0.05).Compared with the control group,the alkaline phosphatase activity,the degree of cell mineralization and the expression of lncRNA H19,BMP2,OCN,OSX,RUNX2 and OPN increased,and the expression of miR-149-5p decreased in the osteogenic group(P＜0.05).Compared with the osteogenesis+si-NC group,the alkaline phosphatase activity,the degree of cell mineralization and the expression of lncRNA H19,BMP2,OCN,OSX,RUNX2 and OPN significantly decreased,and the ex-pression of miR-149-5p increased in the osteogenesis+si-H19 group(P＜0.05).Compared with the osteogenesis+si-H19+miR-inhibitor-NC group,the ALP activity,the degree of cell mineralization and the expression of ln-cRNA H19,BMP2,OCN,OSX,RUNX2 and OPN increased in the osteogenesis+si-H19+miR-inhibitor group.The expression of miR-149-5p significantly decreased(P＜0.05).Compared with the osteogenic+si-H19+pcD-NA-NC group,the alkaline phosphatase activity,the degree of cell mineralization and the expression of BMP2,OCN,OSX,RUNX2 and OPN significantly increased in the osteogenic+si-H19+pcDNA-BMP2 group(P＜0.05).Compared with the sham-operation group and the model group,the alkaline phosphatase activity and the expression of lncRNA H19,BMP2,OCN,OSX,RUNX2 and OPN decreased,and the expression of miR-149-5p increased(P＜0.05).Compared with the model+pcD-null group,the alkaline phosphatase activity and the ex-pression of lncRNA H19,BMP2,OCN,OSX,RUNX2 and OPN significantly increased,and the expression of miR-149-5p decreased in the model+pcD-H19 group(P＜0.05).Conclusion lncRNA H19 promotes osteogenic differentiation of BMSCs and bone regeneration in mice with foot fracture through miR-149-5p/BMP2 axis.关键词
lncRNA H19/成骨分化/足骨骨折/小鼠/骨形成蛋白2/碱性磷酸酶Key words
lncRNA H19/osteogenic differentiation/foot fracture/mice/bone morphogenetic protein 2/alkaline phosphatase分类
医药卫生引用本文复制引用
张怀贵,姑再阿依·买买提,唐卫东..lncRNA H19调控miR-149-5p/BMP2轴对BMSCs的成骨分化及足骨骨折小鼠骨再生的影响[J].安徽医科大学学报,2025,60(6):1059-1068,10.基金项目
新疆维吾尔自治区自然科学基金(编号:2023D01C318) Natural Science Foundation of Xinjiang Uygur Autonomous Region(No.2023D01C318) （编号:2023D01C318）
