胰岛素样生长因子结合蛋白-4对大鼠骨髓间充质干细胞增殖和神经分化的影响  

Impacts of IGFBP4 on proliferation and neural differentiation of bone marrow mesenchymal stem cells in rats

作　　者：李慧文[1] 于树夔[1] 郝飞[1] 牛含虚 徐群渊[1] 段德义[1] 

机构地区：[1]首都医科大学基础医学院神经生物学系北京市神经再生修复研究重点实验室,北京100069

出　　处：《基础医学与临床》2016年第4期513-519,共7页Basic and Clinical Medicine

摘　　要：目的探究胰岛素样生长因子结合蛋白4(IGFBP4)对骨髓间充质干细胞(BMSC)增殖和神经分化的影响。方法通过全骨髓贴壁法分离培养BMSCs,免疫荧光检测IGFBP4在P1、P4、P6、P8和P10 BMSCs的表达。取第4代BMSC细胞,分为单纯培养基组和添加IGFBP4组,用CCK8试剂盒检测单纯培养基组和添加IGFBP 4组吸光度值A450 nm。再将BMSC细胞分为3组:neurocult组、EGF+b FGF组、EGF+b FGF+IGFBP4组,每组连续测定7 d,每天6个平行孔,诱导培养基Neuro Cult培养再分别添加EGF+b FGF和EGF+b FGF+IGFBP4,诱导其向神经祖细胞分化,免疫荧光染色检测Nestin和Sox-2表达,并观察神经球数量和细胞增殖活性。结果成功获得原代大鼠BMSCs,可见CD105、CD44和CD90表达,IGFBP4的表达随培养代数而增加,外源IGFBP4能明显抑制BMSCs的增殖(P<0.05)。经诱导BMSCs形成的神经球样结构表达Nestin和Sox-2,细胞增殖活性升高(P<0.05)。结论 IGFBP4能抑制BMSCs的增殖,促进其向神经祖细胞分化。Objective To explore the effects of insulin-like growth factor binding protein-（ IGFBP4） on the proliferation and neural differentiation of bone marrow mesenchymal stem cell（ BMSC）. Methods BMSCs from SD male rats were isolated by the Whole Marrow Adherence Method and identified using immunofluorescence. IGFBP4 expression in the P1,P4,P6,P8 and P10 BMSCs was detected. P4 BMSCs were divided into two groups,and their proliferation activity was examined by CCK8 assay after treatment with or without exogenous IGFBP4.The P4 BMSCs were further divided into three groups to evaluate their potentials of neural differentiation after treatment with Neuro Cult medium plus EGF ＋ b FGF or EGF ＋ b FGF ＋ IGFBP4. Neural progenitor cells with neurosphere-like structures were identified by immunostaining of Nestin and Sox-2. The number of the induced neurosphere-like structure was counted and the proliferation of neural progenitor cells was assayed. Results BMSCs from SD rats were successfully cultured,and the cells were positive for CD44,CD90 and CD105. The intensity of IGFBP4 expression was increased after passaging,and IGFBP4 could significantly inhibit BMSCproliferation（ P〈0. 05）. BMSCs could be induced to form neural progenitor cells which were immunopositive for Nestin andSox-2. IGFBP4 promoted the differentiation of BMSCs into neural progenitor stem cells,as manifested by an increased size of neurospheres and increased activity of proliferation（ P〈0. 05）. Conclusions IGFBP4 significantly inhibit the proliferation of BMSCs and promote the differentiation of BMSCs into neural progenitor stem cells.

关 键 词：胰岛素样生长因子结合蛋白-4 骨髓间充质干细胞 神经分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]