大鼠神经干细胞中PI3-K/AKT信号转导通路的研究  

Effects of PI3-K/AKT signal transduction pathway in rat neural stem cells

作　　者：赵宇[1] 谢鹏[1] 朱晓峰[2] 

机构地区：[1]重庆医科大学附属第一医院神经内科,重庆400016 [2]佳木斯大学神经科学研究所,佳木斯154007

出　　处：《上海交通大学学报（医学版）》2009年第10期1191-1195,共5页Journal of Shanghai Jiao tong University:Medical Science

基　　金：高等学校博士学科点专项科研基金(20050631012)~~

摘　　要：目的应用PI3-K特异性阻滞剂LY294002孵育大鼠神经干细胞（NSCs）,探讨PI3-K/AKT信号转导通路在NSCs中的作用。方法体外分离、培养E15-16胎鼠NSCs,应用不同浓度（0-40μmol/L）的LY294002孵育NSCs后进行细胞存活（WST-8检测）、增殖（BrdU免疫组织化学鉴定）、分化（βTubulin-Ⅲ免疫组织化学鉴定）和AKT磷酸化蛋白水平（Western Blotting）的检测。结果LY294002对NSCs的存活、增殖和分化的影响呈剂量依赖性。在LY294002较高浓度（25、30、35、40μmol/L）时,NSCs的存活率明显下降（P〈0.05）;LY294002为30、35、40μmol/L时,BrdU阳性细胞数明显少于正常对照组（LY294002 0μmol/L）（P〈0.05）;LY294002为35、40μmol/L时,βTubulin-Ⅲ的阳性细胞数明显少于正常对照组（P〈0.05）。LY294002阻断AKT表达的作用呈剂量依赖性。结论PI3-K/AKT信号转导通路对大鼠NSCs的存活、增殖、分化起重要作用。Objective To incubate rat neural stem cells（NSCs） with selection-specific inhibitor（LY294002） of PI3-K,and explore the effects of PI3-K/AKT signal transduction pathway in NSCs. Methods NSCs derived from E15-16 rats were isolated and cultured.After incubation of NSCs with different concentrations of LY294002（0 to 40 μmol/L）,cell survival（WTS-8 assay）,proliferation（BrdU immunohistochemical identification）,differentiation（β Tubulin-Ⅲimmunohistochemical identification） and AKP phosphorylated protein（Western blotting） were detected. Results The effects of LY294002 on the survival,proliferation and differentiation of NSCs was concentration-dependent.The survival of the NSCs decreased significantly with a higher concentrations of LY294002（25,30,35,40 μmol/L）（P〈0.05）.The numbers of BrdU positive cells with LY294002 of 30,35 and 40 μmol/L were significantly smaller than that of the control（LY294002 0 μmol/L）（P〈0.05）.The numbers of β Tubulin-Ⅲ positive cells with LY294002 of 35 and 40 μmol/L were significantly smaller than that of the control（P〈0.05）.The expression of AKT was blocked by LY294002 in a concentration-dependent manner. Conclusion PI3-K/AKT signal transduction pathway plays a vital role in the survival,proliferation and differentiation of NSCs.

关 键 词：磷脂酰肌醇3激酶 蛋白激酶B 神经干细胞 大鼠 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学]