低氧预处理对心脏成纤维细胞血红素氧化酶1基因表达的影响

Expression of heme oxygenase-1 mRNA after hypoxic preconditioning in neonatal rat cardiac fibroblasts

目的:探讨离体培养的心脏成纤维细胞低氧预处理后血红素氧化酶1(HO-1)mRNA的表达及其经时变化。方法:选用新生W istar大鼠心脏进行成纤维细胞培养,给予低氧预处理(HPC)及低氧/复氧处理(H/R)。测定细胞培养上清液中乳酸脱氢酶(LDH)浓度。自处理后心脏成纤维细胞提取总RNA并进行逆转录扩增合成cDNA。应用即时定量PCR对HO-1mRNA进行定量分析。结果:HPC后HPC组LDH浓度未见显著升高;经过H/R后,H/R组LDH浓度显著高于HPC组(P<0.01)。HPC后HO-1mRNA表达迅速增加,30 m in时达到高峰,此后逐渐下降,24 h基本恢复到基线水平。结论:新生大鼠心脏成纤维细胞低氧预处理可减少低氧/复氧对细胞的损伤,低氧预处理后HO-1mRNA被显著诱导,并在12 h内维持较高水平,24 h后基本恢复至基线水平。

Objective： To investigate the expression of heine oxygenase-1 （ HO-1 ） mRNA and its duration in cardiac fibroblasts after hypoxic preconditioning. Methods： The third passage of cardiac fibroblasts from neonatal Wistar rats was prepared and treated with hypoxia preconditioning （HPC） and hypoxia/reoxygenation （H/R）. The level of lactate dehydrogenase （LDH） in the culture supernatant was measured with cytotoxicity Detection Kit （Roche）. The expression of HO-1 mRNA was determined by real-time polymerase chain reaction （PCR）. Results： The LDH level did not increase significantly after HPC, and after H/R the level was significantly higher than that after HPC （ P 〈 0. 01 ）. The expression of HO-1 mRNA increased after HPC, reached the peak 30 minutes later, declined gradually thereafter, and returned to the baseline level 24 hours later. Conclusion： Hypoxic preconditioning can protect cardiac fibroblasts from the injury induced by sustained hypoxia-reoxygenation. The expression of HO-1 mRNA increases after hypoxic preconditioning, remains high within 12 hours, and returns to the baseline level 24 hours later.