目的探讨不同程度失血大鼠心肌细胞中缺氧诱导因子-1α(hypoxia-inducible factor—1α,HIF-1α)蛋白和mRNA的表达及意义。方法选用清洁级SD大鼠,建立不同程度的失血模型,随机分为假手术对照组(CON)、失血10%(10-BL)组、失血20%(20-BL)组和失血30%(30-BL)组,利用Powerlab八道生理记录仪监测心功能指标,采用RT—PCR检测HIF—1α mRNA在失血后30min、1h大鼠心肌组织中的表达情况,并用免疫组化染色检测HIF—1α在心肌细胞中的分布。结果不同程度失血后,心功能均降低,1h比30min降低明显(P〈0.05);RT—PCR半定量结果显示,与CON组相比,10-BL、20-BL、30-BL组HIF—1αmRNA表达逐渐增高(P〈0.05);免疫组化结果表明,HIF—1α主要分布在心肌细胞细胞质中,在细胞核内也有分布。结论随失血程度增加,时间延长,HIF—1α表达增加,HIF-1α可能参与失血导致的低氧状态下心肌代谢的适应性调节。
Objective This study was to investigate the protein and mRNA expression of hypoxia inducible factor - 1 α ( HIF - 1 α) in rat with different blood loss. Methods SD rats were used to build the animal model of different blood loss. The rats were randomly divided into normal control group, 10 -BL group,20 - BL group and 30 - BL group. Powerlab muhichannel electrophysiolograph was applied to detect the parameter of heart function. Immunohistochemistry, RT - PCR were used to investigate the expression of HIF -1α at 30 min and 1 h after blood loss, respectively. Results Heart function gradually degrade with the level of blood loss , 1 h was lower than 30 rain ( P 〈 0. 05 ). The expression of HIF - 1α could not be detected in the control group. The expression of HIF - 1α mRNA in blood loss group was higher than in control group and gradually increased from 10 -BL, reaching its maximum level on 30 - BL( P 〈 0.05 ). Immunohistochemistry showed that HIF - 1 α protein was mainly present in cytoplasm and/oreytoblast of cardiac muscle. Conclusion These results suggested that the expression of HIF- 1α increased gradually followed by blood loss level and time prolong. HIF- 1α expression may play a key role in the process of myocardial metabolism in hypoxia caused by blood loss in mice.