锌对大鼠骨髓间充质干细胞增殖与向神经样细胞分化的影响  

Effect of zinc on the proliferation and differentiation of bone marrow mesenchymal stem cells into neural-like cells

作　　者：严珺[1] 王永杰[1] 孟春阳[2] 李雷[1] 赵嘉勋 郭贵君 彭川莉 尹飞[2] 郭丽[1] 

机构地区：[1]吉林大学公共卫生学院卫生毒理学教研室,吉林长春130021 [2]吉林大学中日联谊医院骨科,吉林长春130033

出　　处：《中国实验诊断学》2016年第7期1045-1048,共4页Chinese Journal of Laboratory Diagnosis

基　　金：国家自然科学基金(81573067);吉林省科技厅科技发展计划项目资助课题(20130413023GH);吉林省卫生计生科研计划项目(2014Z046);吉林省教育厅"十二五"科学技术研究项目(2015533));大学生创新训练项目(2014A72312)

摘　　要：目的探讨锌对骨髓间充质干细胞(bone marrow mesenchymal stem cells BMSCs)增殖及向神经样细胞诱导分化的作用。方法贴壁法体外分离培养BMSCs,MTT法检测不同浓度的锌处理BMSCs 24h、48h及72h后增殖活性。免疫细胞化学法检测不同浓度的锌处理BMSCs Ki67的表达。BMSCs分空白对照组、缺锌诱导组、正常诱导组、低浓度锌处理组和高浓度锌处理5组。除空白对照组外,其余4组在含有碱性成纤维细胞生长因子(bFGF)、表皮细胞生长因子(EGF)诱导分化培养基中培养5d,维甲酸继续培养14d,诱导BMSCs向神经样细胞分化。倒置显微镜观察细胞生长形态;免疫细胞化学法检测胶质纤维酸性蛋白(GFAP)和神经元特异性烯醇化酶(NSE)的表达。结果MTT结果显示,细胞暴露在锌浓度为200×10-7 mol/L时,不同时间点细胞活力均最强;免疫细胞化学法显示:缺锌组Ki67阳性表达显著低于空白对照组(P<0.05),低浓度锌处理组Ki67阳性表达显著高于空白对照组、缺锌组和高浓度锌处理组(P<0.05);缺锌组、正常诱导组和低浓度锌处理组GFAP和NSE阳性率显著高于空白对照组和高浓度锌处理组(P<0.05),正常诱导组和低浓度锌处理组的GFAP和NSE阳性率显著高于缺锌组(P<0.05),低浓度锌处理组的GFAP和NSE阳性率均显著高于正常诱导组(P<0.05)。结论锌在一定浓度范围内能够提高BMSCs增殖能力及促进BMSCs向神经样细胞分化。Objective To explore the effect of zinc on the proliferation and differentiation of bone marrow mesenchymal stem cells into neural-like cells.Methods BMSCs were isolated by adherence and cultured in vitro.MTT method was used to detect the proliferation activity of BMSCs treated with different concentration zinc in 24 h,48h,72 h.The Ki67 expression in BMSCs administrated by different concentration zinc were detected by immunocytochemical method.The third generation BMSCs were divided into five groups,including blank control group,zinc deficiency group,normal induction group,low concentration zinc treatment group and high concentration zinc treatment.Apart from the blank control group,BMSCs in the other four groups were cultured for 5days in differentiation induction culture medium containing basic fibroblast growth factor（bFGF）and epidermal growth factor（EGF）,then were induced to differentiate into neural-like cells in the culture medium with retinoic acid for 14 days.Cell growth morphology was observed by inverted microscope.The expression of glial fiber acidic protein（GFAP）and neuron specific enzyme（NSE）were detected by immunocytochemistry.Results MTT assay showed cell viability of BMSCs exposed to 200×10-7 mol/L zinc were highest at different time points;immunocytochemistry results showed that Ki67-positive cells percentage of BMSCs in zinc deficiency group was lower than that in blank control group（P〈0.05）,Ki67-positive cells percentage of BMSCs in low zinc treatment group was significantly higher than those in the blank control group,zinc deficiency group and high zinc treatment group（P〈0.05）;GFAP-positive and NSE-positive percentages of BMSCs in zinc deficiency group,the normal induced group and low zinc treatment group were significantly higher than those in blank control group and high zinc treatment group（P〈0.05）,and GFAP-positive and NSE- positive percentages of BMSCs in normal induced group and the low zinc treatment group were significantly higher than that in zinc de

关 键 词：骨髓间充质干细胞 锌 增殖 分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]