目的:观察血红素氧合酶-1(HO-1)体内外对缺血环境下骨髓间充质干细胞(MSCs)的保护作用及梗死后心脏血流动力学的影响。方法:取大鼠骨髓,体外分离扩增培养MSCs;HO-1腺病毒转染MSCs,荧光显微镜下观察绿色荧光蛋白(GFP)的表达;流式细胞仪检测无氧无血清条件培养的HO-1-MSCs、MSCs的凋亡率。建立心梗模型,分别将DAPI-HO-1-MSCs、DAPI-MSCs多点注射到大鼠心脏梗死区周边,对照组注射等量PBS,比较各组MSCs存活率;4周后测定血流动力学指标。结果:HO-1转染的MSCs可稳定高效地表达GFP;在无氧无血清条件下,HO-1-MSCs的凋亡率显著低于MSCs(P〈0.01);细胞移植到梗死后心肌,HO-1-MSCs组存活MSCs计数在第7天明显高于MSCs组(P〈0.05),且血流动力学各项指标较MSCs组明显改善(P〈0.05)。结论:HO-1可在体内、体外实验中提高缺血环境下MSCs的存活率,并改善心肌梗死后血流动力学和心功能,为优化干细胞治疗提供了新思路。
Objective:To investigate the effects of mesenchymal stem cells(MSCs) transfected with heme oxygenase-1(HO-1) gene on its survival under situtation of serum-free and hypoxia in acute myocardial infarction(AMI) heart.Method:MSCs were acquired from the bone marrow of adults rats.They were isolated,purified,cultured,and transfected with Adv-HO-1.The expression of GFP and cell apoptosis were detected by immunofluorescence;Cell apoptosis was detected by FACS respectively.The number of DAPI-positive cells was evaluated as the survival of MSCs.Results:HO-1-MSCs exhibited strong expression of GFP.A lower proportion of apoptosis was observed in HO-1-MSCs compared with MSCs under condition of serum-free and hypoxia(P0.01),and the number of MSCs in AMI heart treated with HO-1-MSCs was significantly higher than those treated with MSCs and PBS(P0.05).Conclusion:HO-1 improve the survival of MSCs treated with serum-free and hypoxia;and HO-1 enhances the survival of MSCs in AMI heart.