间充质干细胞分泌的血管内皮生长因子对心肌细胞的保护作用

Effect of vascular endothelial growth factor from mesenchymal stem cells on anti-apoptosis of H9C2 cells through PI3-K/Akt signal pathway

目的:探索间充质干细胞通过其分泌的血管内皮生长因子对H9C2细胞的保护作用及机制。方法:收集骨髓源间充质干细胞的条件培养基,分别处理培养,将其置于缺氧小室缺氧处理21 h,复氧处理6 h。阻断实验利用VEGFR阻断剂SU5416与间充质干细胞的条件培养基共处理。随后,利用流式细胞术ANNEXIN/PI双标法分析其凋亡变化。Western blotting分析H9C2细胞pAkt的表达。结果:RT-PCR结果显示间充质干细胞表达血管内皮生长因子,Western blotting结果显示间充质干细胞条件培养基增加了H9C2细胞pAkt的水平。AnnexinV/P1分析发现间充质干细胞条件培养基明显降低了H9C2细胞缺氧复氧的凋亡,且这种抗凋亡作用能被VEGFR阻断剂SU5416或PI3-K/Akt途径阻断剂LY294002所阻断。结论:间充质干细胞通过其分泌的血管内皮生长因子通过激活PI3-K/Akt途径保护H9C2细胞。

AIM： To explore the effect of vascular endothelial growth factor （VEGF） from mesenchymal stem cells （MSCs） on anti-apoptosis of H9C2 cells. METHODS： In vitro, condition medium （CM） from MSCs was collected after MSCs were cultured for 2 days. After H9C2 cells were pretreated with VEGFR inhibitor SU5416 （5 p.mol/L） or PI3-K/Akt inhibitor LY294002 （10 panol/L） for 1 h, apoptosis of the ceils was observed under hypoxia with Annexin V and PI double staining by flow eytometry. The expressions of Akt and pAkt in H9c2 cells were detected by Western blotting and expressions of VEGF in MSCs were detected by RT-PCR. RESULTS： RT-PCR showed the expression of VEGF on MSCs, and Western blotting showed increased levels of pAkt protein in H9C2 cells after treatment with CM. CM decreased H9C2 cell apoptosis but the anti-apoptotic effect of the CM was blocked by VEGFR inhibitor SU5416 or PI3-K/Akt inhibitor LY294002 （10 μmol/L）, respectively. CONCLUSION： VEGF from MSCs plays an important role in the protection of H9C2 cells through PI3-K/Akt signal pathway.