生长分化因子5诱导兔骨髓间充质干细胞的软骨分化  

Chondrogenic differentiation of rabbit bone marrow mesenchymal stem cells induced by growth differentiation factor 5

作　　者：李飞非 王布雨 杨治航 董晓宇 邓江 Li Feifei;Wang Buyu;Yang Zhihang;Dong Xiaoyu;Deng Jiang(Third Affiliated Hospital of Zunyi Medical University(First People’s Hospital of Zunyi),Zunyi 563000,Guizhou Province,China)

机构地区：[1]遵义医科大学第三附属医院(遵义市第一人民医院),贵州省遵义市563000

出　　处：《中国组织工程研究》2024年第13期1976-1982,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81660367),项目负责人:邓江;贵州省卫生健康委科学技术基金(gzwkj 2021-236),项目负责人:邓江~~。

摘　　要：背景:生长分化因子5属于转化生长因子超家族成员之一,也是关节发育的最早标志之一,对软骨修复具有重要作用。目的:探讨生长分化因子5诱导骨髓间充质干细胞向软骨分化的作用机制。方法:分离培养兔骨髓间充质干细胞,CCK-8法检测不同质量浓度生长分化因子5对骨髓间充质干细胞增殖活性的影响,RT-PCR检测不同质量浓度生长分化因子5诱导骨髓间充质干细胞成软骨分化相关基因的表达;为进一步探讨生长分化因子5诱导骨髓间充质干细胞成软骨分化的作用机制,加入Wnt/β-catenin信号通路抑制剂XAV-939和激活剂Laduviglusib诱导培养14 d,RT-PCR和Western blot检测软骨相关基因和Wnt/β-catenin信号通路中相关蛋白的表达。结果与结论:①CCK-8结果表明生长分化因子5对骨髓间充质干细胞增殖没有明显影响;②生长分化因子5促进了软骨相关基因Ⅱ型胶原、蛋白聚糖、Sox9的表达,其中以50 ng/mL组软骨相关基因上调明显;③加入Wnt/β-catenin信号通路激活剂Laduviglusib之后,Sox9、β-catenin和Ⅱ型胶原表达上调(P<0.05),加入Wnt/β-catenin信号通路抑制剂XAV939之后,Sox9、β-catenin和Ⅱ型胶原表达下调(P<0.05);④综上,生长分化因子5诱导骨髓间充质干细胞向软骨分化可能与激活Wnt/β-catenin信号通路有关。BACKGROUND:Growth differentiation factor 5 is a member of the transforming growth factor superfamily and one of the earliest markers of joint development.Growth differentiation factor 5 has an important role in cartilage repair.OBJECTIVE:To explore the action mechanism of growth differentiation factor 5-induced chondrogenic differentiation of bone marrow mesenchymal stem cells.METHODS:Rabbit bone marrow mesenchymal stem cells were isolated and cultured.CCK-8 assay was used to detect the effect of different mass concentrations of growth differentiation factor 5 on the proliferation activity of bone marrow mesenchymal stem cells.RT-PCR was utilized to detect the expression of genes related to chondrogenic differentiation of bone marrow mesenchymal stem cells induced by different mass concentrations of growth differentiation factor 5.To further investigate the action mechanism of growth differentiation factor 5-induced chondrogenic differentiation of bone marrow mesenchymal stem cells,we added inhibitor XAV-939 and activator Laduviglusib of Wnt/β-catenin signaling pathways to induce cell culture for 14 days.RT-PCR and western blot assay were performed to detect the expression of cartilage-related genes and Wnt/β-catenin signaling pathway proteins.RESULTS AND CONCLUSION:(1)CCK-8 results showed no significant effect of growth differentiation factor 5 on the proliferation of bone marrow mesenchymal stem cells.(2)Growth differentiation factor 5 promoted the expression of cartilage-related genes type II collagen,aggrecan and Sox9,among which growth differentiation factor 5 induced a significant upregulation of cartilage-related genes in the 50 ng/mL group.(3)Addition of Laduviglusib,an activator of Wnt/β-catenin signaling pathway,upregulated Sox9,β-catenin and type II collagen expression(P<0.05).Addition of XAV939,an inhibitor of Wnt/β-catenin signaling pathway,down-regulated Sox9,β-catenin and type II collagen expression(P<0.05).(4)Taken together,growth differentiation factor 5-induced chondrogenic differentiation

关 键 词：生长分化因子5 骨髓间充质干细胞 软骨分化 软骨细胞 WNT Β-CATENIN 机制 

分 类 号：R459.9[医药卫生—治疗学] R318.19[医药卫生—临床医学] R394.2