17β-雌二醇对骨髓间充质干细胞增殖及成肌分化作用的研究  

Effects of 17β-estradiol on the proliferation and myogenic differentiation of bone marrow mesenchymal stem cells

作　　者：罗新[1] 高绿芬[1] 陈翠平[1] 谢静燕[2] 蒋学风[1] 沈媛[1] 

机构地区：[1]广州暨南大学附属第一医院妇产科,510630 [2]南京医科大学附属南京第一医院妇产科

出　　处：《中国妇产科临床杂志》2010年第5期369-373,共5页Chinese Journal of Clinical Obstetrics and Gynecology

基　　金：广东省科技计划项目(2007B060401054);广东省医学科研基金立项资助课题(A2008364和A2007338);广东省自然科学基金博士科研启动项目(8451063201000290);暨南大学自然科学基金资助项目(200351204003);江苏省卫生厅科研基金项目(H200743)资助

摘　　要：目的探讨17β-雌二醇（17β-E2）对骨髓间充质干细胞（bone marrow mesenchymal stemcells,BMSCs）的增殖和成肌分化的影响。方法利用贴壁法体外分离纯化BMSCs,连续传代3次;MTT法检测在0~1nmol/L浓度范围内17β-E2对BMSCs增殖活性的影响;Hoechst33258染色测定BMSCsDNA含量;免疫荧光法检测添加不同浓度17β-E2诱导培养后BMSCs中骨骼肌分化标志物肌球蛋白重链（MHC）、myogenin和desmin的表达情况。结果 17β-E2可增强BMSCs的增殖活性,且呈浓度依赖关系,最佳促增殖浓度为1nmol/L;1nmol/L17β-E2诱导组与对照组BMSCs DNA含量随培养时间延长均呈增加趋势（P〈0.05）。而两组细胞活性随培养时间的延长均呈下降趋势,17β-E2干预组下降缓慢。17β-E2可增加BMSCs三种骨骼肌特异标志物myogenin、MHC和desmin的表达。结论 17β-E2对BMSCs的增殖具有促进作用,且能增强骨骼肌分化效率。Objective To investigate the effects of 17 beta-estradiol（17β-E2）on myogenic differentiation and proliferation of bone marrow mesenchymal stem cells（BMSCs）,thus to enhance the skeletal differentiation ratio of BMSCs by 5-aza cytidine（5-aza）induction.Methods Bone mesenchymal stem cells were isolated from Sprague-Dawley rats and purified by the ability to adhere to the cultural plastic in vitro.The proliferation effect of 17β-E2 on BMSCs was investigated by MTT method.Hoechst 33258 staining was used to measure the effect of the optinal concentration of 17β-E2 on BMSCs DNA content.Then,the effects of 17β-E2 on BMSCs 5-aza-induced skeletal muscle differentiation were determined by immunofluorescence staining（IF）TUNEL assay.Results 17β-E2 could promote BMSCs proliferation ability in a concentration-dependent manner.The optimal concentration of 17β-E2 was 1nmol/L.17β-E2 could increase BMSCs DNA content with time dependently.Also 17β-E2 could increase the expressions of myogemin,MHC,desmin.Conclusions 17β-E2 could promote BMSCs proliferation,enhance skeletal muscle differentiation induced by 5-aza.

关 键 词：骨髓间充质干细胞 雌激素 增殖 分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]