医学分子生物学杂志2026，Vol.23Issue(5)：523-530,8.DOI:10.3870/j.issn.1672-8009.2026.05.006
慢病毒载体介导miR-195低表达调控BMMSCs成骨分化及血管生成
Effect of Lentiviral Vector-Mediated Knockdown of miR-195 on Osteogenic and Angiogenic Differentiation of BMMSCs
摘要
Abstract
Objective To investigate the regulatory effect and potential mechanism of lentiviral vector-mediated microRNA-195(miR-195)downregulation on osteogenic and angiogenic differentia-tion of bone marrow mesenchymal stem cells(BMMSCs).Methods Human BMMSCs were divided into three groups:blank control group,inhibitor-negative control(NC)group(transfected with len-tiviral NC sequence),and miR-195-inhibitor group(transfected with lentiviral miR-195 inhibitor sequence).The expression and localization of miR-195 were detected by RNA fluorescence in situ hybridization.Cell proliferation was measured by CCK-8 assay.Osteogenic differentiation was evaluated by alkaline phosphatase(ALP)staining,alizarin red staining,and Runt-related transcrip-tion factor 2(Runx2)immunofluorescence.Tube formation assay was performed for angiogenesis assessment.The mRNA levels of vascular endothelial growth factor(VEGF),ALP,Runx2,VEGF receptor 2(VEGFR2),angiopoietin-1(Ang-1),and Wnt/β-catenin pathway-related molecules(Wnt3a,Wnt10b,β-catenin,Osterix)were determined by qRT-PCR.The protein expression of the above pathway and osteogenesis-related molecules was detected by Western blotting.Results Compared with the control groups,the miR-195-inhibitor group showed significantly decreased miR-195 expression(all P<0.05),and markedly increased cell proliferation,ALP activity(4.15±0.52 vs 1.00±0.15),alizarin red positive rate(64.67%±7.78%vs 31.86%±2.24%),Runx2 fluo-rescence intensity(852.38±122.70 vs 121.31±12.16 RFU),and tubular structure length(218.60±24.75 vs 49.80±5.25 µm/field).The mRNA levels of VEGF,ALP,Runx2,VEGFR2,Ang-1,Wnt3a,Wnt10b,β-catenin,and Osterix,as well as the protein levels of Wnt3a,Wnt10b,β-catenin,and Osterix were all significantly upregulated(all P<0.05).There were no significant differences in the above indices between the inhibitor-NC group and the blank control group(P>0.05).Conclusion Lentiviral-mediated miR-195 downregulation synergistically promotes osteo-genic and angiogenic differentiation of BMMSCs via activating the Wnt/β-catenin signaling pathway.关键词
慢病毒载体/微小RNA-195/骨髓间充质干细胞/成骨分化/成血管分化Key words
lentivirus/microRNA-195/bone marrow mesenchymal stem cell/osteogenic differ-entiation/angiogenic differentiation分类
医药卫生引用本文复制引用
丁刘闯,裴培,姬晓炜..慢病毒载体介导miR-195低表达调控BMMSCs成骨分化及血管生成[J].医学分子生物学杂志,2026,23(5):523-530,8.基金项目
"青年科研起航"基金青年项目(No.2024YFY-QKQN-86) This work was supported by a grant from the"Youth Research Start-up"Fund for Young Scholars(No.2024YFY-QKQN-86). （No.2024YFY-QKQN-86）
