骨髓间充质干细胞分泌基质细胞衍生因子1保护心肌细胞  

Stromal cell-derived factor-1 from bone marrow mesenchymal stem cells protects myocardial cells

作　　者：毛洪波 邹赛 唐俊明[2,3] 杨建业[2] 王家宁[2] 孔霞[2] 郭凌郧[2] 郑飞[2] 张蕾[2] 黄永章[2] 

机构地区：[1]湖北省竹溪县人民医院内科,湖北省竹溪县442300 [2]湖北医药学院附属人民医院临床医学研究所,湖北省十堰市442000 [3]湖北医药学院生理学教研室,湖北省十堰市442000

出　　处：《中国组织工程研究》2013年第6期963-968,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81170095;30700306);湖北省卫生厅项目(JX5B24);湖北省教育基金(T201112);郧阳医学院大学生科研项目~~

摘　　要：背景:有研究显示表达CXCR4的干细胞能够沿着基质细胞衍生因子1的浓度梯度迁移到心肌梗死部位再生心肌和血管而改善心脏的功能。目的:探索间充质干细胞通过其分泌的基质细胞衍生因子1对心肌细胞的保护作用。方法:收集培养2d的间充质干细胞条件培养基。在缺氧条件,利用基质细胞衍生因子1受体CXCR4阻断剂AMD3100或PI3-K/Akt途径阻断剂LY294002预处理H9C2细胞后,利用AnnexinV/PI双标法流式细胞术分析间充质干细胞条件培养基作用下H9C2细胞凋亡的变化;Westernblotting分析H9C2细胞磷酸化Akt蛋白的表达;RT-PCR分析间充质干细胞基质细胞衍生因子1的表达。结果与结论:RT-PCR结果显示间充质干细胞表达基质细胞衍生因子1,Westernblotting结果显示间充质干细胞条件培养基增加了H9C2细胞磷酸化Akt蛋白的水平。AnnexinV/PI分析发现间充质干细胞条件培养基明显降低了H9C2细胞缺氧复氧后的凋亡,且这种抗凋亡作用能被CXCR4阻断剂AMD3100或PI3-K/Akt途径阻断剂LY294002所阻断。说明间充质干细胞通过其分泌的基质细胞衍生因子1通过激活PI3-K/Akt途径保护H9C2细胞,增加H9C2细胞的幸存能力。BACKGROUND： Several studies have demonstrated that stem cells expressing CXCR4 can migrate into myocardial infarction region to improve the heart function by regenerating myocardial tissue and vessels under the gradient concentrations of stromal cell-derived factor-1. OBJECTIVE： To investigate the protective effect of stromal cell-derived factor-1 from bone marrow mesenchymal stem cells on myocardial cells. METHODS： Conditioned medium of bone marrow mesenchymal stem cells cultured for 2 days was collected. Under hypoxic condition, H9C2 cells were pretreated with CXCR4 inhibitor AMD3100 （5 mg/mL） or PI3-KJAkt inhibitor LY294002 （10 pM） for 1 hour. After treated with bone marrow mesenchymal stem cell conditioned medium, H9C2 cell apoptosis was analyzed by Annexin V/PI double staining methods. Expression of Akt and pAkt in H9C2 cells was analyzed by Western blotting. Expression of stromal cell-derived factor-1 was analyzed by RT-PCR. RESULTS AND CONCLUSION： RT-PCR results showed that bone marrow mesenchymal stem cells expressed stromal call-derived factor-l. Western blotting results showed that bone marrow mesenchymal stem cell conditioned medium increased pAkt protein level in H9C2 cells. Annexin V/PI analysis showed that bone marrow mesenchymal stem cell conditioned medium significantly decreased apoptosis of H9C2 cells induced by hypoxia/reoxygenation and this anti-apoptotic effect could be blocked by CXCR4 inhibitor AMD3100 or PI3-K/Akt inhibitor LY294002. Stromal cell-derived factor-1 from mesenchymal stem cells plays an important role in the protection of cardiomyocytes through PI3-K/Akt signal pathway.

关 键 词：干细胞 骨髓干细胞 间充质干细胞 H9 C2细胞 心肌细胞 心肌梗死 基质细胞源衍生因子1 凋亡 PI3-K/AKT 国家自然科学基金 干细胞图片文章 

分 类 号：R394.2[医药卫生—医学遗传学]