摘要目的:探究丝裂原活化蛋白激酶（mitogen activated protein kinase，MAPK）通路中ASK1-P38α/JNK在除草醚诱导的先天性膈疝动物模型肺中的表达情况。方法:选取健康成年SPF级SD大鼠，怀孕后通过数字随机法分为空白对照组、膈疝组和膈疝+西地那非干预组3组，通过HE染色检测胎肺发育情况、免疫组织化学定位各组胎肺中的凋亡信号调节激酶1（apoptosis signalregulating kinase 1，ASK1）、丝裂原活化蛋白激酶14（mitogen activated protein kinase 14，P38α）、c-Jun氨基末端激酶（c-Jun N-terminal kinase，JNK）蛋白和实时定量聚合酶链反应（quantificational real time polymerase chain reaction，qRT-PCR）检测各组胎肺的死亡结构域相关蛋白（Death domain associated protein，DAXX）、ASK1、MAPK激酶3（MAP kinase kinase 3，MKK3）、P38α、MAPK激酶4（MAP kinase kinase 4，MKK4）及JNK的mRNA相对表达量。结果:成功获得对照组4只孕鼠50只活胎、膈疝组6只孕鼠84只活胎，干预组4只孕鼠37只活胎。HE染色后与正常组相比，膈疝组的肺明显发育不良及血管重构，干预组明显改善。免疫组织化学显示，ASK1、P38α、JNK在各个组中均有表达，定位在肺叶边缘、气管、气管软骨、周围结缔组织、发育不良全肺叶及部分血管。qRT-PCR显示，ASK1与上游DAXX、下游P38α和JNK均呈正相关（ r=0.778、 P<0.001， r=0.816、 P<0.001和 r=0.284、 P=0.044），ASK1的mRNA表达量升高导致了下游P38α和JNK升高。DAXX中，膈疝组和干预组均较对照组升高（1.28±0.41、1.31±0.30比1.00±0.20， P<0.05）；ASK1中，膈疝组较对照组显著升高（1.51±0.70比1.00±0.23， P<0.05）；MKK3中，干预组较对照组和膈疝组升高（1.21±0.32比1.00±0.18、0.97±0.35， P<0.05）；MKK4中，干预组较对照组和膈疝组显著升高（1.52±0.40比1.00±0.25、1.19±0.38， P<0.05）。 结论:在除草醚诱导的先天性膈疝动物模型肺中，肺发育不良可能与MAPK通路中的ASK1-P38α/JNK上调有关。

abstractsObjective:To explore the expression of ASK1-P38α/JNK in mitogen activated protein kinase (MAPK) pathway in lung of nitrofen-induced congenital diaphragmatic hernia animal model.Methods:After pregnancy, healthy adult specific pathogen free (SPF) Sprague-Dawley (SD) rats were randomly divided into three groups of blank control, diaphragmatic hernia and diaphragmatic hernia plus sildenafil intervention. Hematoxylin-eosin (HE) staining was utilized for detecting the development of fetal lung, immunohistochemical localization of fetal lung protein of Apoptosis signal regulating kinase 1 (ASK1), mitogen activated protein kinase 14 (P38α) and c-Jun N-terminal kinase (JNK). And quantitative real-time polymerase chain reaction (qRT-PCR) were employed for detecting the relative expression of mRNA of death domain associated protein (DAXX), ASK1, MAP kinase kinase 3 (MKK3), P38α, MAP kinase kinase 4 (MKK4) and JNK in fetal lung of three groups.Results:There were 50 live fetuses of 4 pregnant rats in control group, 84 live fetuses of 6 pregnant rats in diaphragmatic hernia group and 37 live fetuses of 4 pregnant rats were successfully obtained in intervention group. HE staining revealed that, as compared with control group, pulmonary dysplasia and vascular remodeling were evident in diaphragmatic hernia group and fetal lung significantly improved in intervention group. Immunohistochemistry indicated that ASK1, P38α and JNK were expressed in all groups. It was located at the edge of lung lobe, trachea, tracheal cartilage, surrounding connective tissue, hypoplastic whole lung lobe and some blood vessels. And qRT-PCR hinted that ASK1 was positively correlated with upstream DAXX, downstream p38 and JNK ( r=0.778, P<0.001, r=0.816, P<0.001, r=0.284, P=0.044). An elevation of ASK1 resulted in the spikes of downstream p38/JNK. In DAXX, diaphragmatic hernia and intervention groups were higher than control group (1.28±0.41, 1.31±0.30 vs. 1.00±0.20, P<0.05); ASK1 was significantly higher in diaphragmatic hernia group than that in control group (1.51±0.70 vs. 1.00±0.23, P<0.05); In MKK3, intervention group was higher than control and diaphragmatic hernia groups (1.21±0.32 vs. 1.00±0.18, 0.97±0.35, P<0.05); In MKK4, intervention group was significantly higher than control and diaphragmatic hernia groups (1.52±0.40 vs. 1.00±0.25, 1.19±0.38, P<0.05). Conclusion:In animal model of nitrofen induced congenital diaphragmatic hernia, pulmonary dysplasia may be correlated with an up-regulation of ASK1-P38α/JNK of MAPK pathway.