骨粘连蛋白促骨髓来源间充质干细胞成骨分化在青少年特发性脊柱侧凸中的作用  

Role of Osteonectin in promoting osteogenic differentiation of bone marrow-derived mesenchymal stem cells in adolescent idiopathic scoliosis

作　　者：赵志蓉[1] 陈涛 陈焕雄 ZHAO Zhi-rong;CHEN Tao;CHEN Huan-xiong(Department of Spinal Surgery,First Affiliated Hospital of Hainan Medical University,Haikou Hainan 570100,China)

机构地区：[1]海南医学院第一附属医院脊柱外科,海南海口570100

出　　处：《局解手术学杂志》2024年第7期561-566,共6页Journal of Regional Anatomy and Operative Surgery

基　　金：国家自然科学基金(82160435)。

摘　　要：目的探究骨骼肌分泌的骨粘连蛋白(Osteonectin)促骨髓来源间充质干细胞(BMSC)成骨分化在青少年特发性脊柱侧凸(AIS)中的作用。方法纳入30例AIS患者,根据X射线片测量的Cobb角,将其分为轻度AIS组和重度AIS组。比较2组患者腰椎和骨骼肌骨密度(BMD)。收集2组患者BMSC,通过流式细胞术检测CD73和CD90蛋白表达,qRT-PCR检测Runx2、Osterix和Osteocalcin mRNAs表达水平,ELISA检测碱性磷酸酶(ALP)活性。收集2组患者骨骼肌,检测Osteonectin mRNA和蛋白表达水平。ELISA检测2组患者血清中Osteonectin水平。体外培养BMSC,分为BMSC组(无特殊处理)和BMSC+Osteonectin组(添加Osteonectin),检测Runx2、Osterix和Osteocalcin mRNAs表达水平及ALP活性。结果与轻度AIS组比较,重度AIS组患者腰椎和骨骼肌BMD降低(P<0.05),BMSC中Runx2、Osterix和Osteocalcin mRNAs表达水平降低(P<0.05),ALP活性降低(P<0.05);骨骼肌中Osteonectin mRNA和蛋白表达水平降低(P<0.05);血清中Osteonectin水平降低(P<0.05)。与BMSC组比较,BMSC+Osteonectin组细胞Runx2、Osterix和Osteocalcin mRNAs表达水平升高(P<0.05),ALP活性升高(P<0.05)。结论AIS患者骨骼肌分泌Osteonectin减少,BMSC的成骨分化能力降低。外源性补充Osteonectin可改善BMSC的成骨分化能力,可能有助于AIS病情的恢复。Objective To investigate the role of Osteonectin secreted by skeletal muscle in promoting the osteogenic differentiation of bone marrow-derived mesenchymal stem cells(BMSC)in adolescent idiopathic scoliosis(AIS).Methods Thirty patients with AIS were included and divided into the mild AIS group and the severe AIS group based on the Cobb angle measured on the X-ray films.The bone mineral density(BMD)of the lumbar spine and skeletal muscles was compared between the two groups.BMSC were collected from patients in the both groups,and the protein expression of CD73 and CD90 were detected by flow cytometry,the mRNA expression levels of Runx2,Osterix,and Osteocalcin were detected by qRT-PCR,and the activity of alkaline phosphatase(ALP)was detected by ELISA.The skeletal muscle from both groups was collected to detect the mRNA and protein expression levels of Osteonectin,and the serum Osteonectin levels in both groups were detected by ELISA.The BMSC cultured in vitro were divided into the BMSC group(no special treatment)and the BMSC+Osteonectin group(addition of Osteonectin),and the mRNA expression levels of Runx2,Osterix and Osteocalcin,as well as ALP activity were detected.Results Compared with the mild AIS group,the severe AIS group had significantly lower BMD of the lumbar spine and skeletal muscle(P<0.05),lower mRNA expression levels of Runx2,Osterix and Osteocalcin(P<0.05)and lower ALP activity in BMSC(P<0.05),lower mRNA and protein expression levels of Osteonectin in skeletal muscle(P<0.05)and lower serum Osteonectin levels(P<0.05).Compared with the BMSC group,the mRNA expression levels of Runx2,Osterix and Osteocalcin,as well as ALP activity in the BMSC+Osteonectin group were significantly increased(P<0.05).Conclusion In patients with AIS,the secretion of Osteonectin by skeletal muscle is reduced,and the osteogenic differentiation ability of BMSC is decreased.Exogenous addition of Osteonectin can improve the osteogenic differentiation of BMSC,which may contribute to the recovery of AIS.

关 键 词：骨髓来源间充质干细胞 成骨分化 青少年特发性脊柱侧凸 骨骼肌 骨粘连蛋白 

分 类 号：R682[医药卫生—骨科学]