内皮型一氧化氮合酶磷酸化在硫化氢调节原位左肺移植后白细胞介素-1β和白细胞介素-10水平中的作用

Exogenous hydrogen sulfide regulates the levels of interleukin-1βand interleukin-10 after orthotopic left lung transplantation via regulating phosphorylated endothelial nitric oxide synthase

目的探讨硫化氢(H2S)通过调节内皮型一氧化氮合酶(eNOS)磷酸化对原位左肺移植引起的肺缺血再灌注损伤模型大鼠移植后IL-1β和IL-10水平的调节作用。方法将24只Sprague-Dawley大鼠随机分入移植组、硫氢化钠(NaHS)组、炔丙基甘氨酸(PPG)组和假手术组,每组6只。前3组大鼠行大鼠左肺原位移植术,冷缺血3h后开放肺门再灌注24h,于再灌注前15min,分别予3组大鼠腹腔内注射0.9%氯化钠溶液、NaHS(14μmol/kg)、PPG(37.5mg/kg);假手术组大鼠仅作开、关胸处理。再灌注24h后,处死大鼠取材。检测eNOS在Ser1117位点的磷酸化水平及其总蛋白水平,以及促炎细胞因子IL-1β和抗炎细胞因子IL-10的水平。结果移植组eNOS(Ser1117)磷酸化水平显著低于NaHS组和假手术组(P值均<0.05),显著高于PPG组(P<0.01)。各组间eNOS总蛋白水平的差异无统计学意义(P值均>0.05)。移植组IL-1β和IL-10水平均显著高于假手术组和NaHS组(P值均<0.01),IL-1β水平显著低于PPG组(P<0.01),IL-10水平显著高于PPG组(P<0.01)。结论 H2S可能通过调节eNOS(Ser1117)磷酸化来调节原位左肺移植后肺组织IL-1β和IL-10的水平。

Objective To investigate the effect of exogenous hydrogen sulfide on IL-1β and IL-10 levels in rats with orthotopic left lung transplantation induced ischemia reperfusion injury （IRI） via regulating phosphorylated endothelial nitric oxide synthase （eNOS）. Methods Twenty-four Sprague-Dawley rats were randomly divided into 4 groups., lung transplantation （LT） group, sodium hydrogen sulfide （NariS） group, propargylglycine （PPG） group and sham operation group （n = 6）. The rats in LT, Naris and PPG groups received orthotropic left lung transplantation with 3 h of cold ischemic period and 24 h of reperfusion period after opening the left pulmonary hilum. Normal saline （NS）, NariS （14 μmol/kg） or PPG （37.5 mg/kg） were injected 15 min before reperfusion in LT, NariS and PPG groups, respectively. The rats in sham operation group underwent sham surgery., the chest was opened and then closed. After 24 h reperfusion, all rats were sacrificed. The phosphorylation of eNOS at its activating site Serl 117 and its total protein levels as well as the levels of IL-1β and IL-10 were tested. Results The level of eNOS （Ser1117） phosphrylation in LT group was significantly lower than those in NariS and sham operation groups （both P〈0.05）, but significantly higher than that of PPG group （P〈0.01）. There were no significant difference in the level of total eNOS protein between groups （all P〈0.05）. The levels of IL-1β and IL-10 in LT group were significantly higher than those in sham operation group and Naris group （all P〈0.01）. The level of IL-1β in LT group was significantly lower than that in PPG group （P〈0.01）, but the level of IL-10 in LT group was significantly higher than that in PPG group （P〈0.01）. Conclusion The exogenous hydrogen sulfide may regulate the levels of IL-1β and IL-10 after orthotopic left lung transplantation via the regulation of the level of phosphorylated eNOS （Ser1117）.