低氧对培养心肌细胞表达和分泌血管内皮生长因子的影响

目的　探讨低氧对心肌细胞血管内皮生长因子 (VEGF)表达的影响。方法　采用Kuwabara等并加以改进的方法对心肌细胞进行低氧培养 ,并于低氧培养的即刻、2 4、4 8h分别测定培养液中的氧分压。使用逆转录聚合酶链反应 (RT PCR)、免疫组织化学和酶联免疫吸附测定 (ELISA)法分别检测低氧培养的不同时间点VEGFmRNA及VEGF蛋白的表达和分泌。结果　与常规培养相比 ,简易低氧培养培养液氧分压降至 5 8mmHg ,并于 3个时间点保持这一水平 (P >0 .0 5 )。从mRNA水平和蛋白水平我们检测到VEGF表达的增加 ,于低氧培养 2 4h达峰值 ,培养液中的峰值浓度为 (72 4 .6 7± 5 6 .87)pg ml。结论　低氧促进心肌细胞表达VEGFmRNA、VEGF蛋白和分泌该蛋白

The effect of hypoxia on the expression and secretion of vascular endothelial growth factor in cultured cardiomyocytes

Objective To study the effect of hypoxia on the vascular endothelial growth factor (VEGF)expression in cardiomyocytes.Methods Cardiomyocytes were cultured in hypoxic condition.PO 2 of cultured medium was tested at 0,24 and 48 hours after hypoxia.VEGFmRNA and its protein were examined respectively by RT PCR,immunohistochemistry and ELISA at different time points of hypoxic culture.Results PO 2 was about 58 mmHg in hypoxic culture and continued 48 hours(P>0.05).The expression of VEGF was increased at the levels of mRNA and protein,and reached a peak at 24 hour of hypoxic culture.The peak level of VEGF in culture medium was (724.67±56.87) pg/ml.Conclusion Hypoxia promotes the expression and secretion of VEGF in cultured cardiomyocytes.