BACKGROUND Degenerative disc disease(DDD)is characterized by the loss of nucleus pulposus cells(NPCs).Inducing differentiation of bone marrow mesenchymal stem cells(MSCs)into NPCs has emerged as a novel therapeutic st...BACKGROUND Degenerative disc disease(DDD)is characterized by the loss of nucleus pulposus cells(NPCs).Inducing differentiation of bone marrow mesenchymal stem cells(MSCs)into NPCs has emerged as a novel therapeutic strategy for DDD.However,the efficiency of MSC differentiation and the underlying mechanisms remain to be fully elucidated.AIM To investigate the role and mechanism of miR-365 in promoting the differentiation of MSCs into NPCs for DDD treatment.METHODS In vitro,the effects of miR-365 on MSC proliferation,apoptosis,and differentiation were assessed by cell counting kit-8 assay,flow cytometry,and quantitative realtime polymerase chain reaction(qRT-PCR).In vivo,the expression levels of miR-365,HIF-1α,Sox9,Kdm6a,and HOXA9 in the spinal cord of rats with spinal cord injury were determined by qRT-PCR and Western blot.RESULTS In vitro,miR-365 significantly promoted MSC proliferation and inhibited MSC apoptosis.The expression levels of glycosaminoglycans,proteoglycan,and type 2 collagen were significantly increased with miR-365 ectopic expression.In vivo,the expression levels of miR-365,HIF-1α,Sox9,and Kdm6a were significantly increased,whereas HOXA9 was remarkably decreased.Mechanically,miR-365 inhibited HOXA9 expression by directly binding to its 3’untranslated region.HOXA9 could inhibit HIF-1αexpression by binding to the Hif-1αpromoter,thereby affecting the expression levels of Sox9 and Kdm6a.Moreover,HOXA9 knockdown significantly reversed the differentiation of MSCs into NPCs induced by miR-365.CONCLUSION miR-365 promotes HOXA9-mediated differentiation of MSCs into NPCs by interacting with HIF-1αand may serve as a potential target for DDD treatment.展开更多
基金Supported by Academic Subject Boosting Plan in Shanghai Fourth People's Hospital Affiliated to Tongji University School of Medicine,No.SY-XKZT-2019-3002Academic Project from the Health Commission of Hongkou District,Shanghai,No.2202-25Clinical Key Specialty Construction Unit from The Health Commission of Hongkou District,Shanghai,No.HKLCZD2024B03.
文摘BACKGROUND Degenerative disc disease(DDD)is characterized by the loss of nucleus pulposus cells(NPCs).Inducing differentiation of bone marrow mesenchymal stem cells(MSCs)into NPCs has emerged as a novel therapeutic strategy for DDD.However,the efficiency of MSC differentiation and the underlying mechanisms remain to be fully elucidated.AIM To investigate the role and mechanism of miR-365 in promoting the differentiation of MSCs into NPCs for DDD treatment.METHODS In vitro,the effects of miR-365 on MSC proliferation,apoptosis,and differentiation were assessed by cell counting kit-8 assay,flow cytometry,and quantitative realtime polymerase chain reaction(qRT-PCR).In vivo,the expression levels of miR-365,HIF-1α,Sox9,Kdm6a,and HOXA9 in the spinal cord of rats with spinal cord injury were determined by qRT-PCR and Western blot.RESULTS In vitro,miR-365 significantly promoted MSC proliferation and inhibited MSC apoptosis.The expression levels of glycosaminoglycans,proteoglycan,and type 2 collagen were significantly increased with miR-365 ectopic expression.In vivo,the expression levels of miR-365,HIF-1α,Sox9,and Kdm6a were significantly increased,whereas HOXA9 was remarkably decreased.Mechanically,miR-365 inhibited HOXA9 expression by directly binding to its 3’untranslated region.HOXA9 could inhibit HIF-1αexpression by binding to the Hif-1αpromoter,thereby affecting the expression levels of Sox9 and Kdm6a.Moreover,HOXA9 knockdown significantly reversed the differentiation of MSCs into NPCs induced by miR-365.CONCLUSION miR-365 promotes HOXA9-mediated differentiation of MSCs into NPCs by interacting with HIF-1αand may serve as a potential target for DDD treatment.
