细胞外信号调节激酶1/2在慢性肺疾病新生大鼠肺成纤维细胞中的研究

摘要

Objective To investigate dynamic changes of extracellular signal regulated protein kinase (ERK) 1/2 in lung fibroblast of newborn rats with chronic lung disease (CLD) caused by hyperoxia.Methods Full-term newborn rats were randomly divided into two groups:air-exposed group and hyperoxia - exposed with 90％ oxygen group.Rats were sacrificed separately 3 d,7 d and 14 days after exposure to air or 90％ oxygen. Then lung fibroblasts of rats were isolated and primarily cultured. By using Immunocystochemistry,Western-blot and RT-PCR methods,the levels of ERK1/2 protein and expressions of ERK1/2 mRNA were measured. Results The levels of p-ERK1/2 protein in lung fibroblast in the hyperoxia group were significant higher on the 7th day and 14th day after exposure to 90％ oxygen compared with those in the air-exposed group (P ＜0.01 ).And the levels of total ERK1/2 protein and expressions of ERK1/2 mRNA did not change noticeably and were not significantly different between two groups (P ＞0.05 ).Conclusions The activation of phosphorated ERK1/2 may lead to lung fibrosis caused by hyperoxia in newborn rats.%目的 探讨持续吸入高氧致慢性肺疾病新生大鼠肺成纤维细胞中细胞外信号调节激酶(ERK) 1/2蛋白信号表达的动态变化.方法 足月新生鼠生后12 h内分别持续吸人体积分数90％的高氧和空气,于3、7、14 d随机处死动物后,进行肺成纤维细胞的原代培养,应用免疫组化、Western-blot及real-time PCR方法检测ERK1/2蛋白及mRNA表达.结果 免疫组化及Western blot结果显示,高氧7、14 d时高氧组肺成纤维细胞p-ERK1/2蛋白的表达均明显高于同时间点对照组(P＜0.01).Western blot结果同时显示各组间ERK1/2总蛋白的表达差异无统计学意义.real-time PCR结果表明各组间ERK1、ERK2 mRNA水平差异无统计学意义(P＞0.05).结论 ERK1/2蛋白水平磷酸化活化参与了高氧致新生大鼠慢性肺疾病中肺纤维化的发生.