锌指蛋白3调节骨形态发生蛋白2诱导C3H10T1/2细胞的成骨分化  

Schnurri3 regulates osteogenic differentiation of C3H10T1/2 cells induced by bone morphogenetic protein 2

作　　者：谢映春 许文娟 李豫皖 Xie Yingchun;Xu Wenjuan;Li Yuwan(Department of Blood Transfusion,Sichuan Academy of Medical Sciences·Sichuan Provincial People’s Hospital,Chengdu 610072,Sichuan Province,China;Chongqing Emergency Medical Center,Chongqing 400014,China;Department of Sports Medicine,Third Hospital of Peking University,Institute of Sports Medicine of Peking University,Beijing Key Laboratory of Sports Injuries,Beijing 100191,China)

机构地区：[1]四川省医学科学院·四川省人民医院输血科,四川省成都市610072 [2]重庆市急救医疗中心,重庆市400014 [3]北京大学第三医院运动医学科,北京大学运动医学研究所,运动医学关节伤病北京市重点实验室,北京市100191

出　　处：《中国组织工程研究》2023年第33期5270-5276,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:骨形态发生蛋白2是参与骨骼生长发育和修复的重要因子,近年来的研究发现锌指蛋白3参与调节转化生长因子β信号的转导,该研究以此为基础开展研究。目的:探讨抑制锌指蛋白3对骨形态发生蛋白2诱导C3H10T1/2细胞成骨分化的影响。方法:以小鼠间充质干细胞系C3H10T1/2细胞为研究对象,设置绿色荧光蛋白、骨形态发生蛋白2、锌指蛋白3、骨形态发生蛋白2+锌指蛋白3重组腺病毒转染组,增强骨形态发生蛋白2表达,抑制锌指蛋白3表达。采用碱性磷酸酶染色检测碱性磷酸酶的表达;RT-qPCR检测骨形态发生蛋白2、锌指蛋白3及成骨、成血管标志物mRNA转录水平;免疫组织化学染色检测Ⅰ型胶原、血管内皮生长因子及内皮黏蛋白表达水平;茜素红染色及半定量分析检测钙盐沉积水平;裸鼠皮下成骨实验检测异位骨块形成情况。结果与结论:(1)骨形态发生蛋白2重组腺病毒能诱导C3H10T1/2细胞的成骨分化,与骨形态发生蛋白重组腺病毒组比较,骨形态发生蛋白2+锌指蛋白3重组腺病毒组成骨标志物碱性磷酸酶、Ⅰ型胶原、成骨细胞特异性转录因子、骨钙素、Runt相关转录因子2及成血管标志物神经轴突导向因子、血管内皮生长因子、血管性血友病因子、促血管生成素及内皮黏蛋白mRNA转录水平显著升高(P<0.001),同时显著增强Ⅰ型胶原、血管内皮生长因子、内皮黏蛋白在蛋白水平的表达(P<0.05)和钙盐沉积水平(P<0.05);(2)骨形态发生蛋白2及锌指蛋白3单独作用C3H10T1/2细胞均能在裸鼠皮下形成异位骨块,骨形态发生蛋白2联合锌指蛋白3能形成更显著的异位骨块;(3)结果表明,抑制锌指蛋白3可显著增强骨形态发生蛋白2诱导C3H10T1/2细胞成骨分化;抑制锌指蛋白3可通过促进成血管因子mRNA及蛋白的表达调控血管生成。BACKGROUND:This study is based on the evidence that bone morphogenetic protein 2 plays an important role in osteogenesis and bone repair,and Schnurri3 can regulate the transforming growth factor-βsignal transduction.OBJECTIVE:To investigate the effect of inhibiting Schnurri3 on bone morphogenetic protein 2-induced osteogenic differentiation of C3H10T1/2 cells.METHODS:Mouse mesenchymal stem cell line C3H10T1/2 cells were used as research materials.Four groups were set in this study:adenovirus-green fluorescent protein,adenovirus-bone morphogenetic protein 2,adenovirus-Simshn3 and adenovirus-bone morphogenetic protein 2+adenovirus-Simshn3.The expression of bone morphogenetic protein 2 was enhanced and the expression of Schnurri3 was inhibited.Alkaline phosphatase staining was used to detect the expression of alkaline phosphatase.The mRNA transcriptional levels of bone morphogenetic protein 2,Schnurri3,osteogenic and angiogenic markers were determined by RT-qPCR.Immunohistochemical staining was used to detect the expression of type I collagen,vascular endothelial growth factor and endothelial mucin.Alizarin red staining and semiquantitative analysis were used to detect calcium salt deposition levels.Subcutaneous stem cell implantation in nude mice was used to measure ectopic bone.RESULTS AND CONCLUSION:(1)Recombinant adenovirus-bone morphogenetic protein 2 could induce osteogenic differentiation of C3H10T1/2 cells.Compared with the adenovirus-bone morphogenetic protein 2 group,mRNA transcription levels of osteogenic markers alkaline phosphatase,type I collagen,osteoblast-specific transcription factor,osteocalcin,Runt related transcription factor 2 and angiogenic markers neurite guidance factor,vascular endothelial growth factor,von Willebrand factor,angiopoietin and endothelial mucin were significantly increased in the adenovirus-bone morphogenetic protein 2+adenovirus-Simshn3 group(P<0.001).Simultaneously,it significantly enhanced the expression of type I collagen,vascular endothelial growth factor and endothelial

关 键 词：间充质干细胞 骨形态发生蛋白2 锌指蛋白3 成骨分化 成血管因子 

分 类 号：R459.9[医药卫生—治疗学] R318.5[医药卫生—临床医学] R681