CAJ | 학술논문

目的探讨环磷酸腺苷-蛋白激酶A (cAMP-PKA)信号通路对大鼠深低温缺血再灌注后肺组织水通道蛋白-5(AQP5)的调控及其与肺损伤的关系.方法将28只Wistar大鼠随机分为假手术组(sham组)、深低温缺血再灌注损伤组(I/R组)、H89组和forskolin组,每组7只.I/R组大鼠体表降温至深低温后阻断左下肺门30 min后开放、复温；sham组只开胸但不降温阻断左下肺门；H89组和forskolin组分别于实验前2d腹腔注射H89和forskolin,其余操作同I/R组；测量左下肺组织湿重/干重比值(W/D比值)同时观察组织学变化,实时定量聚合酶链反应(Real-time PCR)以及Westem blot法检测左下肺组织中AQP5 mRNA和蛋白表达.结果 I/R组与sham组比较,肺组织AQP5 mRNA表达减少(0.67±0.15,P＜0.01)、AQP5蛋白表达减少(0.47±0.09,P＜0.01),W/D比值增高(5.27±0.92,P＜0.01),肺组织形态及结构明显受损；forskolin干预组与I/R组比较,肺组织AQP5 mRNA表达增加(0.83 ±0.30,P＜0.01)、AQP5蛋白表达增加(0.89±0.07,P＜0.01)、W/D比值减低(3.98±0.45,P＜0.01),肺组织病理形态学改变轻于I/R组,与sham组比较,AQP5表达减少,但差异无统计学意义；H89干预组与I/R组和sham组比较,肺组织AQP5 mRNA表达减少(0.53±0.11,P＜0.01)、AQP5蛋白表达减少(0.31±0.03,P＜0.01),W/D比值增高(6.13 ±0.78,P＜0.01),肺组织形态及结构受损最为明显.结论深低温缺血再灌注肺组织中AQP5表达下调且与肺损伤程度相关,cAMP-PKA信号通路可能参与AQP5的调控.
목적 탐토배린산선감-단백격매A (cAMP-PKA)신호통로대대서심저온결혈재관주후폐조직수통도단백-5(AQP5)적조공급기여폐손상적관계.방법 장28지Wistar대서수궤분위가수술조(sham조)、심저온결혈재관주손상조(I/R조)、H89조화forskolin조,매조7지.I/R조대서체표강온지심저온후조단좌하폐문30 min후개방、복온；sham조지개흉단불강온조단좌하폐문；H89조화forskolin조분별우실험전2d복강주사H89화forskolin,기여조작동I/R조；측량좌하폐조직습중/간중비치(W/D비치)동시관찰조직학변화,실시정량취합매련반응(Real-time PCR)이급Westem blot법검측좌하폐조직중AQP5 mRNA화단백표체.결과 I/R조여sham조비교,폐조직AQP5 mRNA표체감소(0.67±0.15,P＜0.01)、AQP5단백표체감소(0.47±0.09,P＜0.01),W/D비치증고(5.27±0.92,P＜0.01),폐조직형태급결구명현수손；forskolin간예조여I/R조비교,폐조직AQP5 mRNA표체증가(0.83 ±0.30,P＜0.01)、AQP5단백표체증가(0.89±0.07,P＜0.01)、W/D비치감저(3.98±0.45,P＜0.01),폐조직병리형태학개변경우I/R조,여sham조비교,AQP5표체감소,단차이무통계학의의；H89간예조여I/R조화sham조비교,폐조직AQP5 mRNA표체감소(0.53±0.11,P＜0.01)、AQP5단백표체감소(0.31±0.03,P＜0.01),W/D비치증고(6.13 ±0.78,P＜0.01),폐조직형태급결구수손최위명현.결론 심저온결혈재관주폐조직중AQP5표체하조차여폐손상정도상관,cAMP-PKA신호통로가능삼여AQP5적조공.
Objective To discuss the regulatory effect of cyclic adenosine monophosphate-protein kinase A (cAMP-PKA) signaling pathway on the apuaporin 5 (AQP5) expression in a rat model of lung ischemia reperfusion after deep hypothermia and its relation to lung injury.Methods Twenty-eight Wistar rats were randomly divided into sham group,ischemia-reperfusion group (I/R group),H89 group,and Forskolin group.The left lower hilus pulmonis of L/R group was blocked for 30 min after its body temperature cooled to deep hypothermia and soon opened and rewarmed.The rats in the sham group only undergone thoracotomy without cooling and blocking.The rats in the H89 group and Forskolin group were intraperitoneally injected with H89 and Forskolin 2 days before experiment,respectively.The rest procedures were the same to those in the I/R group.The lung specimens were pathologically observed and wet/dry weight ratio was measured.Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting methods were applied to detect the expression of AQP5 in all groups.Results As compared with sham group,the expression of AQP5 mRNA was reduced (0.67 ± 0.15,P ＜ 0.01),the expression of AQP5 protein declined (0.47 ± 0.09,P ＜ 0.01),W/D ratio increased (5.27 ± 0.92,P ＜ 0.01),the morphology and structure of lung tissue was significandy impaired in the I/R group.As compared with I/R group,the expression of AQP5 mRNA was increased (0.83 ± 0.30,P ＜0.01),the expression of AQP5 protein increased (0.89 ± 0.07,P ＜ 0.01),W/D ratio was reduced (3.98 ± 0.45,P ＜ 0.01),the injury of lung tissue was significantly alleviated in the Forskolin group.There was no statistically significant difference between sham group and Forskolin group.As compared with I/R group and sham group,the AQP5 mRNA expression was decreased (0.53-0.11,P ＜ 0.01),the AQP5 protein expression decreased (0.31 ±0.03,P ＜ 0.0 1),W/D ratio increased (6.13 ±0.78,P ＜ 0.01),and injury of lung tissue was significantly aggravated in H89 group.Conclusion AQP5 was down-regulated in acute lung injury by deep hypothermia ischemia-reperfusion.cAMP-PKA signaling pathway may participate in the regulation of AQP5.