丝裂原活化蛋白激酶信号对高氧性肺损伤水通道蛋白5的调控机制  

Regulation mechanism of MAP kinases on aquaporin 5 mRNA and protein expression in hyperoxic lung injury

作　　者：周璟[1] 谭利平[1] 许峰[2] 方芳[3] 

机构地区：[1]重庆医科大学附属儿童医院急诊科 [2]重庆医科大学附属儿童医院PICU [3]儿童发育疾病研究省部共建教育部重点实验室、重庆市儿童发育重大疾病诊治与预防国际科技合作基地、儿科学重庆市重点实验室,重庆400014

出　　处：《重庆医科大学学报》2014年第9期1231-1236,共6页Journal of Chongqing Medical University

基　　金：中华儿科杂志百利儿科科研基金资助项目;重庆医科大学校级课题资助项目

摘　　要：目的:探讨丝裂原活化蛋白激酶(mitogen activated protein kinases,MAPKs)信号通路三亚族ERK、p38、JNK对高氧性肺损伤异常表达水通道蛋白5(aquaporin 5,AQP5)的分子调控机制。方法:制备高氧肺损伤动物模型,通过不同信号途径抑制剂干预。随机将Wistar实验大鼠分为8组(n=6):空气对照组(A)、高氧组(H)、高氧+ERK抑制剂PD98059组(H+PD)、高氧+p38抑制剂SB203580组(H+SB)、高氧+JNK抑制剂SP600125组(H+SP)及空气+抑制剂对照组(A+PD、A+SB、A+SP),采用蛋白质免疫印迹法检测AQP5蛋白表达,real-time PCR检测AQP5 mRNA。结果:Western blot结果显示:A组AQP5蛋白呈较高表达,H组AQP5表达减弱(P=0.000),而H+SB、H+SP组较H组AQP5蛋白表达明显升高(P=0.000),二者中尤以H+SB组其蛋白上调更明显,H+PD组与H组比较无统计学性差异(P=0.737);real-time PCR结果显示:H组AQP5 mRNA表达较空气对照组降低(P=0.000),H+SB、H+SP与H组比较AQP5 mRNA表达升高(P=0.000),二者中尤以H+SB组其mRNA上调更明显,但二者与A组比较,差异仍有统计学意义(P=0.000),而H+PD干预组与H组比较,未见肺组织AQP5 mRNA表达的明显变化(P=0.796)。A组与A+抑制剂对照组(A+PD、A+SB、A+SP)在AQP5蛋白及mRNA表达上均无明显差异。结论:抑制剂SB203580及SP600125在高氧暴露后能增加AQP5蛋白和mRNA表达,因而推测MAPKs通路中p38、JNK可能参与了调控高氧肺损伤AQP5基因表达的下调过程。Objective：To investigate regulation mechanism of mitogen activated protein kinases（MAPKs）signaling pathway on Aquaporin 5（AQP5）expression disorder in hyperoxic lung injury（HLI）. Methods：Forty-eight Wistar rats aged three weeks old were randomly divided into 8 groups（n=6）：room-air group（A）,hyperoxia group（H）,hyperoxia ＋ ERK inhibitor PD98059（H＋PD）,hyperoxia＋ p38 inhibitor SB203580（H＋SB）,hyperoxia ＋ JNK inhibitor SP600125（H＋SP）,and the corresponding inhibitors control groups（A＋PD,A＋SB,A＋SP）. The rats in hyperoxia groups were kept in cabinet with high concentration of oxygen（≥95%）,and those in air groups were kept in room air. AQP5 protein expression in lung tissue was detected by Western blot. AQP5 mRNA expression was determined by real-time PCR. Results：Lung injury developed in hyperoxia exposure groups,as evidenced by severe alveolar edema,inflammatory cell infiltration,RBC leakage and alveolar structure destruction. The protein level of AQP5 detected by Western blot was down- regulated in H group and H＋ inhibitors（H＋PD/SB/SP）groups than in A group（P=0.000）,but the expression of AQP5 protein was markedly higher in H＋SB and H＋SP groups than in H group（P=0.000）,and this change was more significant in H＋SB group.There was no difference in AQP5 protein expression between H group and H＋PD group（P=0.737）. The mRNA level of AQP5 was decreased in H group but was increased in H ＋SB,H ＋SP and H groups compared with that of A group（P=0.000）. However,there was no change in AQP5 mRNA level between H＋PD and H group（P=0.796）. There was no significant change in AQP5 protein and mRNA expression between A group and A＋inhibitors（A＋PD/SB/SP）groups. Conclusion：Hyperioxia decreases AQP5 protein and mRNA expression while SB203580 and SP600125 up-regulates AQP5 gene expression. Inhibition of p38 and JNK pathway blocks the effect of hyperoxia on AQP5 expression. These data show that p38 and JNK MAP kinases may invol

关 键 词：急性肺损伤 高氧 水通道蛋白5 丝裂原活化蛋白激酶通路 

分 类 号：R563[医药卫生—呼吸系统]