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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: Studies have shown that CXCR4-expressing stem cells improve cardiac function by migrating myocardial and blood vessels along the concentration gradient of stromal cell-derived factor 1 to sites of myocardial infarction. Objective: To explore the protective effects of mesenchymal stem cells derived from stromal cell derived factor 1 on cardiomyocytes. Methods: Mesenchymal stem cell conditioned medium was collected for 2 days. Hypoxic conditions, the use of stromal cell-derived factor 1 receptor CXCR4 blocker AMD3100 or PI3-K / Akt pathway blocker LY294002 pretreatment H9C2 cells, using Annexin V / PI double-labeled flow cytometry of mesenchyme The changes of H9C2 cell apoptosis under the condition of stem cell conditioned medium, the expression of phosphorylated Akt protein in H9C2 cells by Western blotting, and the expression of stromal cell derived factor-1 in mesenchymal stem cells by RT-PCR. RESULTS AND CONCLUSION: RT-PCR results showed that MSCs express stromal cell-derived factor 1. Western blotting showed that MSCs conditioned medium increased phosphorylated Akt protein in H9C2 cells. AnnexinV / PI analysis showed that MSCs conditioned medium significantly reduced the apoptosis of H9C2 cells after hypoxia-reoxygenation, and this anti-apoptotic effect could be blocked by the CXCR4 blocker, AMD3100 or PI3-K / Akt pathway blocker Blocked by LY294002. It indicates that mesenchymal stem cells can secrete HIF-1 through the PI3-K / Akt pathway to protect H9C2 cells and increase the survivability of H9C2 cells.