Osteonectin促进骨髓来源间充质干细胞成骨分化的机制及其与青少年特发性脊柱侧凸的相关性研究  

Mechanism of Osteonectin Promoting Osteogenic Differentiation of Bone Marrow-derived Mesenchymal Stem Cells and Its Correlation with Adolescent Idiopathic Scoliosis

作　　者：赵志蓉[1] 陈涛 陈焕雄 ZHAO Zhirong;CHEN Tao;CHEN Huanxiong(Department of Spinal Surgery,the First Affiliated Hospital of Hainan Medical University,Haikou 570100,China)

机构地区：[1]海南医学院第一附属医院脊柱外科,海口570100

出　　处：《转化医学杂志》2024年第2期177-182,共6页Translational Medicine Journal

基　　金：国家自然科学基金项目(82160435)。

摘　　要：目的 探究Osteonectin在促进骨髓间充质干细胞(BMSCs)成骨分化的分子机制,以及Osteonectin在青少年特发性脊柱侧凸(AIS)中的作用。方法 将2020年5月—2022年5月就诊的20例AIS及健康体检20例分别作为AIS组和健康组。使用qRT-PCR技术测定2组BMSCs中Osteonectin、BMP2以及Wnt/β-catenin信号通路相关基因(Tcf7、Lef1)和成骨分化相关基因(Runx2、Osteocalcin)的表达水平。通过X线检测AIS患者的Cobb角度,并分析上述基因表达水平与Cobb角度的相关性。在BMSCs中,加入Osteonectin和BMP2抑制剂Noggin,并分为对照组、Osteonectin组和Osteonectin+Noggin组。通过qRT-PCR检测BMSCs中BMP2、Tcf7、Lef1、Runx2和Osteocalcin mRNA表达水平。采用ChIP-qPCR检测β-catenin在Runx2和Osteocalcin基因转录起始位(TSS)的富集水平。结果 与健康组比较,AIS组BMSCs中Osteonectin、BMP2、Tcf7、Lef1、Runx2和Osteocalcin mRNA表达水平均降低(P<0.05);Osteonectin、BMP2、Tcf7、Lef1、Runx2和Osteocalcin mRNA表达水平与AIS患者Cobb角度呈负相关(r=-0.466 3、-0.369 1、-0.272 7、-0.543 4、-0.606 5、-0.343 3,P<0.05,P<0.01)。与对照组比较,Osteonectin组BMSCs中BMP2、Tcf7、Lef1、Runx2和Osteocalcin mRNA表达水平升高,且β-catenin在Runx2和Osteocalcin mRNA TSS富集水平增加(P<0.05)。相比Osteonectin组,Osteonectin+Noggin组BMSCs中Tcf7、Lef1、Runx2和Osteocalcin mRNA表达水平降低,β-catenin在Runx2和Osteocalcin mRNA TSS富集水平降低(P<0.05)。结论 在AIS患者中,BMSCs中Osteonectin、BMP2-Wnt/β-catenin的表达以及成骨分化水平与AIS疾病的发展呈负相关。Osteonectin通过激活BMP2-Wnt/β-catenin信号通路,促使β-catenin转录激活成骨分化相关基因,从而促进BMSCs的成骨分化。Objective To explore the molecular mechanism of Osteonectin(ON)in promoting osteogenic differentiation of bone marrow-derived mesenchymal stem cells(BMSCs)and the role of ON in the development of adolescent idiopathic scoliosis(AIS).Methods Twenty AIS patients and 20 healthy subjects who underwent physical examination from May 2020 to May 2022 were enrolled as AIS group and healthy groups respectively.The expression levels of ON,BMP2 and Wnt/β-catenin signaling pathway-related genes(Tcf7,Lef1)and osteogenic differentiation-related genes(Runx2,Osteocalcin)in BMSCs of the two groups were determined by qRT-PCR.The Cobb angle of AIS patients was detected by X-ray,and the correlation between the above gene expression levels and the Cobb angle was analyzed.BMSCs were added with ON and BMP2 inhibitor Noggin,and divided into control group,ON group,and ON+Noggin group.The expression levels of BMP2,Tcf7,Lef1,Runx2 and Osteocalcin mRNA in BMSCs were detected by qRT-PCR.The enrichment level ofβ-catenin in Runx2 and Osteocalcin gene transcription start sites(TSS)was detected by ChIP-qPCR.Results Compared with healthy group,the expression levels of ON,BMP2,Tcf7,Lef1,Runx2 and Osteocalcin mRNA in BMSCs in AIS group were decreased(P<0.05).The expression levels of ON,BMP2,Tcf7,Lef1,Runx2 and Osteocalcin were negatively correlated with the Cobb angle in AIS patients(r=-0.4663,-0.3691,-0.2727,-0.5434,-0.6065,-0.3433,P<0.05,P<0.01).Compared with the control group,the expression levels of BMP2,Tcf7,Lef1,Runx2 and Osteocalcin mRNA in BMSCs in ON group were increased,and the TSS enrichment levels of Runx2 and Osteocalcin mRNA inβ-catenin group were increased(P<0.05).Compared with the ON group,the expression levels of Tcf7,Lef1,Runx2 and Osteocalcin genes in BMSCs in the ON+Noggin group were decreased,and the TSS enrichment level ofβ-catenin in Runx2 and Osteocalcin genes was also decreased(P<0.05).Conclusion In AIS patients,the expression of ON,BMP2-Wnt/β-catenin in BMSCs,and the level of osteogenic differentiation are negatively

关 键 词：青少年特发性脊柱侧凸 骨结合素 骨粘连蛋白 骨髓来源间充质干细胞 染色质免疫共沉淀 成骨分化 COBB角 BMP2-Wnt/β-catenin信号通路 

分 类 号：R682.3[医药卫生—骨科学]