p38丝裂素活化蛋白激酶信号传导通路对低氧预处理后心脏成纤维细胞血红素氧化酶-1基因表达的影响

p38 MAPK signaling pathway involved in the expression of HO-1 mRNA following hypoxic preconditioning in neonatal cardiac fibroblasts

目的:探讨p38丝裂素活化蛋白激酶(MAPK)信号传导对低氧预处理(HPC)的细胞保护作用以及HPC后血红素氧化酶-1(HO-1)mRNA表达的影响。方法:选用新生Wistar大鼠心脏进行成纤维细胞培养,给予HPC及低氧/复氧处理(HR)。药物处理组在HPC和HR处理过程中向培养液中加入SB203580。测定细胞培养上清液中乳酸脱氢酶(LDH)浓度和细胞HO-1 mRNA表达量。结果:低氧/复氧后,HPC/SB组LDH浓度显著高于HPC/NSB组(P<0.05)。HR组低氧/复氧后各亚组LDH浓度均显著升高(P<0.01,P<0.01);低氧/复氧处理前HPC组中NSB亚组于低氧预处理后HO-1 mRNA表达显著上升(P<0.001),低氧/复氧处理后其表达有所下降;SB亚组其表达也显著上升(P<0.01)。HR组中各亚组于低氧/复氧处理后HO-1 mRNA表达也显著上升(P<0.01)。结论:对培养的心脏成纤维细胞给予低氧预处理能够减轻其后低氧/复氧对细胞的损伤,低氧预处理较低氧/复氧处理更能够显著诱导心脏成纤维细胞HO-1 mRNA的表达。

Objective：To investigate whether p38MAPK signaling pathway involved in the protection of hypoxic preconditioning（HPC） to cardiac fibroblasts from hypocia-reoxygenation（HR） injury and the expression of HO-1mRNA in cardiac fibroblasts following HPC.Methods：Cardiac fibroblasts were prepared from neonatal Wistar rats in the third passage at confluence.HPC and HR treatment were applied to them.Cell treatment with SB203580 were performed by adding it into the culture medium during HPC and HR.The released LDH-activity of the culture supernatant was measured.HO-1 mRNA levels were examined by real-time PCR.Results：LDH activity in HPC/SB group was significantly higher than NSB group after HPC/HR（P 〈 0.05）.In HR group,LDH activity after HR were significantly higher in both NSB and SB subgroup（P 〈 0.01,P 〈 0.01）,In HPC group,the transcription induction of HO-1 elevated most significantly in NSB subgroup after HPC（P 〈 0.001）,then,it decreased slightly after HPC/HR（P 〈 0.001）;expression of HO-1 also increased significantly（P 〈 0.01） in SB subgroup,In HR group,HO-1 mRNA increased significantly after HR（P 〈 0.01）.Conclusion：In summary,hypoxic preconditioning can reduce a subsequent hypoxic/reoxygenation-induced injury in cardiac fibroblasts.This protection can be inhibited by the administration of SB203580,a selective inhibitor of p38 MAPK.Elevation of HO-1mRNA induction following HPC was more significant than that following HR,this augmented expression could also be suppressed by SB203580.