【摘要】 目的:观察缺氧诱导因子-1对心肌细胞缺氧复氧损伤的保护作用。方法:取SD新生大鼠(1～3 d)分离培养成心肌细胞;在培养的第3 d将心肌细胞随机分为3组(每组重复6次):正常对照组、H/R组、DMOG+H/R组;建立缺氧/复氧损伤模型,以脯氨酸羟化酶的抑制剂甲基异二酰基甘氨酸进行干预;采用酶免疫法检测各组心肌细胞培养上清液中的肌钙蛋白I含量,采用RT-PCR法检测各组缺氧诱导因子-1α及下游因子血红素氧合酶-1、血管内皮生长因子、葡萄糖转运蛋白1 mRNA水平。结果:培养4～6 h的乳鼠心肌细胞开始贴壁生长,12～24 h明显增殖,3～4 d后细胞融合成片;心肌细胞由圆形变为梭形、星形、多角形,并出现自发性节律性搏动;与正常对照组相比,H/R组、DMOG+H/R组的肌钙蛋白I含量明显增高(P<0.01),且缺氧诱导因子-1α、下游因子血红素氧合酶-1、血管内皮生长因子、葡萄糖转运蛋白1水平明显增高(P<0.01,P<0.05);与H/R组相比,DMOG+H/R组心肌细胞缺氧诱导因子-1α、下游因子血红素氧合酶-1、血管内皮生长因子、葡萄糖转运蛋白1水平明显增高而肌钙蛋白I含量明显降低(P<0.05)。结论:本研究从细胞水平证实缺氧诱导因子-1的稳定表达对心肌细胞缺氧/复氧损伤具有保护作用。更多还原

【Abstract】 Objective To investigate the protective role of hypoxia inducible factor-1 in cultured cardiocytes with hypoxia/reoxygenation.Methods Cultured cardiocytes of neonatal SD rats(1～3 d) were randomly divided into control group,H/R group and DMOG+H/R group.Models of cardiocytes in vivo were interfered with Prolyl 4-hydroxylase inhibitor dimethyloxaloxalyl glycine.The expression levels of hypoxia inducible factor-1α,hemeoxygenase-1,vascular endothelial growth factor,glucose transporter 1mRNA and the influence of them on the levels of cadiac troponin Ⅰ were determined with ELISA and RT-PCR methods.Results Cardiocytes began to adhere and grow after 4～6 hours,to prolife obviously after 12～24 hours,and to converge together after 3～4 days.The cardiocytes showed sphericalm,fusiform and polygon shape in the visual field of inverted microscope.All the cells stretched out parapodium and beat spontaneously and rhythmically.The expression levels of hypoxia inducible factor-1α,hemeoxygenase-1,vascular endothelial growth factor,glucose transporter 1 mRNA and the contents of cardiac troponin Ⅰ increased in the cardiocytes from H/R injury and DMOG+H/R(P<0.05).The expression levels of hypoxia inducible factor-1α,hemeoxygenase-1,vascular endothelial growth factor and glucose transporter 1 mRNA increased in the cardiocytes pretreated with dimethyloxaloxalyl glycine as compared with the cardiocytes from hypoxi-reoxygenation injury(P<0.01,P<0.05),while the levels of cardiac troponin Ⅰ decreased significantly(P<0.05).Conclusion It is suggested that the upregulated expression of hypoxia inducible factor-1α may be involved in the process of protection of cultured cardiocytes in hypoxia/reoxygenation injury.更多还原