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目的探讨GATA-4基因对骨髓间充质干细胞(MSCs)在低氧环境下存活能力的影响。方法将GATA-4基因转染大鼠MSCs,以转染空质粒的MSCs做对照,通过蛋白印迹法检测GATA-4转染效果。模拟体内心肌梗死氧化应激损伤,在低氧环境下培养48 h,观察MSCs形态变化。通过MTT细胞增殖测定、膜联蛋白V-PE凋亡检测和乳酸脱氢酶(LDH)释放评价MSCs存活能力。结果在低氧环境下,GATA-4基因修饰组与转染空质粒的对照组相比,MTT摄取增加、膜联蛋白V阳性细胞数降低、LDH释放减少,差异有统计学意义(P<0.05)。结论转录因子GATA-4可以增加MSCs抗缺氧能力,增加细胞存活;为MSCs移植治疗缺血性心肌病、增加细胞存活提供新的方法。
Objective To investigate the effect of GATA-4 gene on the viability of bone marrow mesenchymal stem cells (MSCs) in hypoxic environment. METHODS: GATA-4 gene was transfected into rat MSCs and transfected into empty MSCs as control. The effect of GATA-4 transfection was detected by Western blotting. The injury of oxidative stress was simulated in vivo and cultured in hypoxic environment for 48 h. Morphological changes of MSCs were observed. The viability of MSCs was evaluated by MTT cell proliferation assay, Annexin V-PE apoptosis assay and lactate dehydrogenase (LDH) release. Results Compared with the control group transfected with empty plasmid, the GATA-4 gene modified group had higher MTT uptake, lower annexin V-positive cell number and lower LDH release in hypoxia environment, with significant difference (P <0.05 ). Conclusion The transcription factor GATA-4 can increase the anti-hypoxia ability of MSCs and increase cell survival. It provides a new method for transplanting MSCs to treat ischemic cardiomyopathy and increase cell survival.