血管内皮生长因子对骨髓源间充质干细胞增殖的影响及其信号机制  

Effect of vascular endothelial growth factor on bone marrow-derived mesenchymal stem cell proliferation and the signaling mechanism

作　　者：张俊[1,2,3] 谢珊珊[1,2,3] 韩晓霞[1,2,3] 任金涛[1,2,3] 吕富冉[4] 唐俊明[1,3] 郑飞[1] 郭凌郧[1] 杨建业[1] 孔霞[1] 张蕾[1] 黄永章[1] 王家宁[1] 

机构地区：[1]湖北医药学院附属人民医院临床医学研究所,山东临沂276000 [2]湖北医药学院生理学教研室,山东临沂276000 [3]湖北医药学院国际药护学院,湖北十堰442000 [4]湖北医药学院临沂市煤炭疗养院,山东临沂276000

出　　处：《南方医科大学学报》2011年第10期1697-1700,1713,共5页Journal of Southern Medical University

基　　金：国家自然科学基金(81170095;30700306);湖北省卫生厅基金(JX5B24);湖北省教育基金(T2011012);郧阳医学院大学生科研项目~~

摘　　要：目的探索血管内皮生长因子(VEGF)对间充质干细胞(MSC)增殖的影响及其信号机制。方法采用经典的全骨髓贴壁法培养MSC,通过成骨、成脂肪等多向诱导分化以及流式细胞仪分析其表面标记(CD45、CD34、CD90、CD29)等鉴定MSC特征;以P3MSC为实验材料,采用MTT法分析VEGF对MSC增殖的影响。随后以50 nmol/L Wortmannin、50μmol/LPD98059、30μmol/L SB203580、10μmol/L H89、20μmol/L Y27632、1μmol/L Rapamycin、10μmol/L Straurosporine、6nmol/L Gǒ6976、50μmol/L Pseudo Z等分别处理P3MSC,观察VEGF影响MSC增殖的信号机制。结果培养的P3MSC呈现出CD90、CD29强阳性,具有成骨、成脂肪等多向分化能力;VEGF促进MSC增殖,Y27632、PD98059、SB203580、Gǒ6976、Straurosporine可抑制VEGF促进MSC增殖的效应。结论 VEGF引起的MSC增殖效应与AKT-mTOR-PKC途径密切相关,且PKCα处于中间环节。Objective To observe the effect of vascular endothelial growth factor（VEGF） on bone marrow-derived mesenchymal stem cell（MSC） proliferation and explore the signaling mechanism involved.Methods MSC culture was performed following the classical whole bone marrow adhering method.The characteristics of MSC were identified by induction of multi-lineage differentiation and flow cytometry for surface marker analysis（CD34,CD45,CD29,and CD90）.Following the addition of 50 nmol/L wortmannin,50 μmol/L PD98059,30 μmol/L SB203580,10 μmol/L H89,20 μmol/L Y27632,1 μmol/L rapamycin,10 μmol/L straurosporine,6 nmol/L Gǒ6976,or 50 μmol/L Pseudo Z inhibitors in the cell culture,the MSC were treated with 20 ng/ml VEGF and the changes of the cell proliferation rate was measured with MTT assay.Results Cultured MSC were capable of multi-linage differentiation and did not express VEGF-R,CD29 or CD90.Treatment with 20 ng/ml VEGF obviously promoted MSC proliferation,and this effect was inhibited partially by p38 mitogen-activated protein kinase（MAPK） inhibitor rapamycin,PD98059,SB203580,Gǒ6976,and straurosporine.Conclusion VEGF promotes MSC proliferation in close relation to the AKT-PKC pathway,in which PKC signal pathway may play the central role.

关 键 词：粒细胞集落刺激因子 间充质干细胞 增殖 蛋白激酶C 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]