极度低氧条件下基质细胞衍生因子-1α过表达对间充质干细胞迁移的影响  

Effects of stromal cell-derived factor-1αoverexpression on migration of mesenchymal stem cells under extreme hypoxia

作　　者：王辉 徐亮亮 孙大明[2] 孟一 张成龙 许海甲 宁宇 李章华 WANG Hui;XU Liangliang;SUN Daming;MENG Yi;ZHANG Chenglong;XU Haijia;NING Yu;LI Zhanghua(Department of Orthopaedics,Tongren Hospital of Wuhan University,Hubei Province,Wuhan 430000,China;Graduate School,Wuhan Sports University,Hubei Province,Wuhan 430000,China;Department of Orthopaedics,Xiangyang Hospital of Traditional Chinese Medicine,Hubei Province,Xiangyang 441000,China)

机构地区：[1]武汉大学同仁医院骨科,湖北武汉430000 [2]武汉体育学院研究生院,湖北武汉430000 [3]湖北省襄阳市中医医院骨科,湖北襄阳441000

出　　处：《中国医药导报》2022年第16期5-10,共6页China Medical Herald

基　　金：国家自然科学基金资助项目(81472103);湖北省自然科学基金项目(2020CFB369);湖北省卫生健康委面上项目(WJ2021M010);湖北省武汉市医学科研项目(WX18 M01、WX20D91);湖北省武汉市科技局应用基础前沿项目(2019020701011471);湖北省襄阳市科技局医疗卫生领域科技计划项目(2020YL25)。

摘　　要：目的研究极度低氧条件下基质细胞衍生因子-1α(SDF-1α)过表达对间充质干细胞(MSCs)迁移的影响。方法将来源于两只普通级雄性日本大耳白兔[2周龄,体重(250±30)g]的MSCs分为空白组(不进行干预),AMD3100组[加入SDF-1α受体(CXCR4)特异性抑制剂AMD3100],SDF-1α腺病毒(Ad-SDF-1α)组(进行Ad-SDF-1α转染)和Ad-SDF-1α+AMD3100组(进行Ad-SDF-1α转染并添加AMD3100)。四组在极度低氧(1%)条件下培养48 h,通过Transwell实验检测其迁移能力,采用实时荧光定量PCR及Western blot检测迁移信号轴缺氧诱导因子-1(HIF-1α)、血管内皮生长因子(VEGF)、CXCR4的mRNA及蛋白表达情况。结果MSCs贴壁后为梭形、多角形,呈克隆式生长,Ad-SDF-1α转染后可观察到均一且明亮的绿色荧光。AMD3100组光密度(OD)值低于空白组,Ad-SDF-1α组OD值高于空白组,Ad-SDF-1α+AMD3100组OD值高于AMD3100组且低于Ad-SDF-1α组,差异均有统计学意义(P<0.05)。AMD3100组VEGF、CXCR4 mRNA表达低于空白组,Ad-SDF-1α组HIF-1α、VEGF、CXCR4 mRNA表达均高于空白组,差异有统计学意义(P<0.05或P<0.01)。Ad-SDF-1α+AMD3100组VEGF、CXCR4 mRNA表达均高于AMD3100组,HIF-1α、VEGF、CXCR4 mRNA表达均低于Ad-SDF-1α组,差异有统计学意义(P<0.05或P<0.01)。AMD3100组HIF-1α、VEGF、CXCR4蛋白表达低于空白组,Ad-SDF-1α组HIF-1α、VEGF、CXCR4蛋白表达均高于空白组,Ad-SDF-1α+AMD3100组HIF-1α、VEGF、CXCR4蛋白表达均高于AMD3100组且均低于Ad-SDF-1α组,差异有统计学意义(P<0.05或P<0.01)。结论极度低氧条件下SDF-1α过表达仍可促进MSCs迁移,并一定程度上可逆转AMD3100的抑制效应。Objective To investigate the effects of stromal cell-derived factor-1α(SDF-1α)overexpression on migration of mesenchymal stem cells(MSCs)under extreme hypoxia.Methods MSCs derived from two normal male Japanese white rabbits(two weeks of age,body weight[250±30]g)were divided into blank group(no intervention),AMD3100 group(added CXC subfamily receptor 4[CXCR4]specific inhibitor AMD3100),SDF-1αadenovirus(Ad-SDF-1α)group(Ad-SDF-1αtransfected),and Ad-SDF-1α+AMD3100 group(Ad-SDF-1αtransfected and added AMD3100).The four groups were cultured under extreme hypoxia(1%)for 48 h,and the changes of migration ability were detected by Transwell experiment.The mRNA and protein expressions of HIF-1α,vascular endothelial growth factor(VEGF),and CXCR4 were detected by real-time quantitative PCR and Western blot.Results MSCs adhered to the wall in spindle and polygonal shape,and showed clonal growth.Uniform and bright green fluorescence could be observed after transfection with Ad-SDF-1α.The optical density(OD)value of AMD3100 group was lower than that of blank group,OD value of Ad-SDF-1αgroup was higher than that of blank group,OD value of Ad-SDF-1α+AMD3100 group was higher than that of AMD3100 group and lower than that of Ad-SDF-1αgroup,the differences were statistically significant(P<0.05).The mRNA expressions of VEGF and CXCR4 in AMD3100 group were lower than those in blank group,while the mRNA expressions of HIF-1α,VEGF,and CXCR4 in Ad-SDF-1αgroup were higher than those in blank group,the differences were statistically significant(P<0.05 or P<0.01).The mRNA expressions of VEGF and CXCR4 in Ad-SDF-1α+AMD3100 group were higher than those in AMD3100 group,while the mRNA expressions of HIF-1α,VEGF,and CXCR4 were lower than those in Ad-SDF-1αgroup,the differences were statistically significant(P<0.05 or P<0.01).The protein expressions of HIF-1α,VEGF,and CXCR4 in AMD3100 group were lower than those in blank group,while the protein expressions of HIF-1α,VEGF,and CXCR4 in Ad-SDF-1αgroup were higher than those in b

关 键 词：极度低氧 间充质干细胞 基质细胞衍生因子-1Α 迁移 

分 类 号：R318[医药卫生—生物医学工程]