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

目的：探讨离体培养的心脏成纤维细胞低氧预处理后血红紊氧化酶1（HO-1）mRNA的表达及其经时变化。方法：选用新生Wistar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，30min时达到高峰，此后逐渐下降，24h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12h内维持较高水平，24h后基本恢复至基线水平。

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

Objective: To investigate the expression of heme 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.