miR-212-3p regulates senescence of bone marrow mesenchymal stem cells by targeting MAPK3
- VernacularTitle:miR-212-3p靶向MAPK3调控骨髓间充质干细胞的衰老
- Author:
Liying ZHONG
1
;
Shundong LI
1
;
Cong WANG
1
Author Information
- Publication Type:Journal Article
- Keywords: bone marrow mesenchymal stem cell; cell senescence; miR-212-3p; mitogen-activated protein kinase 3; β-galactosidase
- From: Chinese Journal of Tissue Engineering Research 2025;29(13):2690-2697
- CountryChina
- Language:Chinese
- Abstract: BACKGROUND:Bone marrow mesenchymal stem cells in patients with osteoporosis show significant senescence and decreased activity and osteogenic differentiation.miR-212-3p inhibits osteogenic differentiation of human bone marrow mesenchymal stem cells.However,its regulation of senescence of bone marrow mesenchymal stem cells and its mechanism remain unclear.OBJECTIVE:To investigate the effect of miR-212-3p on senescence of bone marrow mesenchymal stem cells by targeting mitogen-activated protein kinase 3 (MAPK3) and its mechanism.METHODS:Rat bone marrow mesenchymal stem cells were isolated and cultured in vitro,and the third generation was collected for the following experiments:(1) Cultured in two groups:The control group was added with complete culture medium,and the model group was added with complete culture medium containing H2O2.After 72 hours of culture,β-galactosidase activity,miR-212-3p and MAPK3 mRNA expression,as well as MAPK3,p16,and p21 protein expression were detected.(2) Cultured in three groups:control group,inhibitor control group,and miR-212-3p inhibitor group.After transfection for 24 hours,miR-212-3p,mRNA and protein expression of MAPK3 were detected.(3) Dual luciferase reporter gene combined with qRT-PCR and western blot assay were used to verify the targeting regulation of miR-212-3p and MAPK3.(4) Cultured in different groups:control inhibitor group,miR-212-3p inhibitor group,miR-212-3p inhibitor+interference control group,and miR-212-3p inhibitor+MAPK3 interference group.After transfection for 24 hours,MAPK protein and mRNA expression levels in cells were detected.They were divided into control group,H2O2 group,H2O2+control inhibitor group,H2O2+miR-212-3p inhibitor group,H2O2+miR-212-3p inhibitor+interference control group,and H2O2+miR-212-3p inhibitor+MAPK3 interference group.Cells were transfected for 24 hours and then cultured with H2O2 for 72 hours.Aging-related β-galactosidase activity and p16 and p21 protein expression were detected.RESULTS AND CONCLUSION:(1) Compared with the control group,β-galactosidase activity,miR-212-3p mRNA expression and p16,p21 protein expression were increased in the model group (P<0.05),while MAPK3 mRNA and protein expression levels were decreased (P<0.05).(2) Compared with the control group,the mRNA expression of miR-212-3p was decreased (P<0.05),and the mRNA and protein expression levels of MAPK3 were increased (P<0.05) in miR-212-3p inhibitor group.(3) Double luciferase reporter gene experiment confirmed that MAPK3 was the downstream target gene of miR-212-3p.(4) Compared with the control inhibitor group,the mRNA and protein expression levels of MAPK3 were increased in miR-212-3p inhibitor group (P<0.05).Compared with the miR-212-3p inhibitor group,the mRNA and protein expression levels of MAPK3 in the miR-212-3p inhibitor+MAPK3 interference group were decreased (P<0.05).Compared with H2O2+control inhibitor group,β-galactosidase activity in H2O2+miR-212-3p inhibitor group was decreased (P<0.05).Compared with H2O2+miR-212-3p inhibitor group,β-galactosidase activity in H2O2+miR-212-3p inhibitor+MAPK3 interference group was higher than that in H2O2+miR-212-3p inhibitor group (P<0.05).Compared with the H2O2+control inhibitor group,the protein expression levels of p16 and p21 in the H2O2+miR-212-3p inhibitor group were decreased (P<0.05).Compared with H2O2+miR-212-3p inhibitor group,the protein expression levels of p16 and p21 in H2O2+miR-212-3p inhibitor+MAPK3 interference group were increased (P<0.05).(5) To conclude,downregulation of miR-212-3p inhibits the senescence of rat bone marrow mesenchymal stem cells,and its mechanism of action may be achieved by targeting up-regulation of MAPK3 expression.
