GDNF在精原干细胞中对H19和A-myb表达的调控研究  

Role of GDNF in regulation of expression of H19 and A-myb in spermatogonial stem cells

作　　者：查巍巍[1] 曹静萍 张迪[1] 冯立新[1] 程金科[1] 

机构地区：[1]上海交通大学医学院干细胞研究室,上海200025 [2]中国科学院上海生命科学研究院/上海交通大学医学院健康科学研究所,上海200025

出　　处：《上海交通大学学报（医学版）》2011年第12期1715-1718,1723,共5页Journal of Shanghai Jiao tong University:Medical Science

摘　　要：目的通过研究胶质细胞神经源性营养因子(GDNF)对H19和A-myb表达的调控,探讨GDNF调节精原干细胞自我更新的分子机制。方法采用混合酶消化和差速贴壁法从出生后6 d的小鼠睾丸中分离、鉴定并培养精原干细胞。用GDNF分别刺激精原干细胞和经磷酸肌醇3激酶/蛋白激酶B(PI3K/Akt)通路抑制剂LY294002预处理的精原干细胞后,采用RT-PCR技术检测细胞H19和A-myb的表达。结果相关鉴定分析证实分离培养的细胞为精原干细胞。RT-PCR检测结果显示:GDNF刺激下的精原干细胞的H19和A-myb表达均显著上调,而经LY294002预处理的精原干细胞在GDNF刺激下其H19和A-myb的表达无明显变化。结论在体外培养的小鼠精原干细胞中,GDNF通过PI3K/Akt信号通路上调H19和A-myb的表达。Objective To investigate the regulation of expression of H19 and A-myb by glial cell line-derived neurotrophic factor（GDNF）,and explore the molecular mechanism of self-renewal of spermatogonial stem cells regulated by GDNF.MethodsSpermatogonial stem cells were isolated,identified and cultured from testes of 6-day-old mice with mixed enzyme digestion and differential plating procedure.Spermatogonial stem cells and spermatogonial stem cells pretreated with LY294002,inhibitor of phosphotylinosital 3 kinase/protein kinase B（PI3K/Akt）,were stimulated with GDNF,and the expression of H19 and A-myb in spermatogonial stem cells was detected by RT-PCR.ResultsSpermatogonial stem cells were successfully isolated and cultured.The expression of H19 and A-myb was upregulated in spermatogonial stem cells after stimulation with GDNF,while there was no significant change in the expression of H19 and A-myb in spermatogonial stem cells after pretreatment with LY294002.ConclusionGDNF can upregulate the expression of H19 and A-myb via PI3K/AKT signal pathway in cultured mouse spermatogonial stem cells.

关 键 词：胶质细胞源性神经营养因子 精原干细胞 H19 A-myb 磷酸肌醇3激酶/蛋白激酶B 

分 类 号：Q132.1[生物学—普通生物学]