Bmp2影响脂肪干细胞成骨能力的研究  

Study on the effect of Bmp2 on the osteogenic ability of adipose stem cells

作　　者：赖文涛 赵志军[2,3] 张春阳 Lai Wentao;Zhao Zhijun;Zhang Chunyang(Baotou Medical College,Inner Mongolia University of Science and Technology,Baotou 014010;Dept of Neurosurgery,The First Affiliated Hospital of Baotou Medical College of Inner Mongolia University of Science and Technology,Baotou 014010;Engineering Technology Center for Bone Tissue Regeneration and Injury Repair in Inner Mongolia Autonomous Region,Baotou 014010)

机构地区：[1]内蒙古科技大学包头医学院,包头014010 [2]内蒙古科技大学包头医学院第一附属医院神经外科,包头014010 [3]内蒙古自治区骨组织再生与损伤修复工程技术中心,包头014010

出　　处：《安徽医科大学学报》2023年第6期963-969,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81660214、81960238);内蒙古自治区自然科学基金(编号:2020MS08013);内蒙古自治区高等学校科学研究项目(编号:NJZZ20168);内蒙古自治区科技成果转化项目(编号:NM2019BT052);包头市医学重点学科建设项目(编号:BWXK201910);包头医学院科学研究基金(编号:BYJJ-YF-2018004);国家临床重点专科建设项目(编号:Z155080000004)。

摘　　要：目的探究在大鼠脂肪干细胞内过表达骨形态发生蛋白2(Bmp2)后成骨基因的表达情况以及成骨能力的变化。方法通过电转染Bmp2至大鼠脂肪干细胞内,以空白组作为对照,分为空白对照组(Control组)、空载组(Vector组)、成骨诱导组(Inducement组)、过表达组(Bmp2组),采用Western blot、qRT-PCR及免疫荧光对比成骨基因表达情况,采用碱性磷酸酶染色与茜素红染色观察硫化钴沉淀及钙盐沉积情况。结果Western blot与qRT-PCR检测显示,与Control组相比,Bmp2组的Bmp2、Bmp4、骨桥蛋白(OPN)、骨钙蛋白(OCN)、矮小相关转录因子2(Runx2)、磷酸化大鼠信号转导分子1/5(P-Smad1/5)、远端缺失同源框2(Dlx2)、骨涎蛋白(BSP)表达显著增高(P<0.05),而Smad1/5的表达情况差异无统计学意义;免疫荧光检测显示,与Control组相比,Bmp2组的OPN、Runx2荧光强度显著增高(P<0.05);茜素红染色及碱性磷酸酶染色结果显示,与Control组相比,Bmp2组细胞周围钙盐沉积增多,硫化钴沉淀增多,碱性磷酸酶活性增强。结论过表达Bmp2增强了脂肪干细胞的成骨基因表达,提高了其成骨能力。Objective To investigate the changes of osteogenic genes and osteogenic capacity after overexpression of bone morphogenetic protein 2(Bmp2)in rat adipose stem cells.Methods Bmp2 was transfected into rat adipose stem cells by electrotransfection,and the blank group was used as control.The cells were divided into blank group(Control group),empty load group(Vector group),osteogenesis induction group(Inducement group),and overexpression group(Bmp2 group).Western blot,qRT-PCR and immunofluorescence were used to compare the expression of osteogenic genes,and alkaline phosphatase staining and alizarin red staining were used to observe cobalt sulfide precipitation and calcium salt deposition.Results Western blot and qRT-PCR showed that the expression of Bmp2,Bmp4,OPN,OCN,Runx2,P-Smad1/5,Dlx2,and BSP was significantly higher compared to the Control group(P<0.05),while the expression of Smad1/5 was not statistically different.Immunofluorescence showed that the fluorescence intensity of OPN and Runx2 was significantly higher compared to the Control group(P<0.05).The alizarin red staining and alkaline phosphatase staining showed that the Bmp2 group had increased pericellular calcium salt deposition,increased cobalt sulfide precipitation,and enhanced alkaline phosphatase activity compared to the Control group.Conclusion Overexpression of Bmp2 enhances osteogenic gene expression in adipose stem cells and improves their osteogenic ability.

关 键 词：脂肪干细胞 BMP2 成骨分化 

分 类 号：R651.15[医药卫生—外科学]