艾司氯胺酮对肝脏缺血-再灌注模型幼鼠肺损伤的保护作用

Protective effect of esketamine on lung injury induced by hepatic ischemia-reperfusion in young mice models

目的 探讨艾司氯胺酮对肝脏缺血-再灌注(I-R)模型幼鼠肺损伤的保护作用及其可能机制。方法 将18只C57幼鼠随机分为三组,每组6只。C组为假手术组;I-R组和S组采用肝脏缺血6 h建立肝脏I-R模型。I-R组为模型对照组;S组建模前经尾静脉注射艾司氯胺酮10 mg/kg。I-R组和S组于肝脏I-R 6 h后,C组于关腹6 h后取血清、支气管肺泡灌洗液(BALF)及肺组织。采用HE染色观察肺组织病理学改变,ELISA法检测血清高迁移率族蛋白B1(HMGB1)水平及BALF中TNF-α、IL-1β水平,硫代巴比妥酸法测定肺组织丙二醛(MDA)浓度,羟胺法测定肺组织超氧化物歧化酶(SOD)活性,免疫组织化学染色法检测肺组织HMGB1表达,Western blot法检测肺组织磷脂酰肌醇3-激酶(PI3K)、Akt、哺乳动物雷帕霉素靶蛋白(mTOR)的蛋白表达。结果 与C组相比,I-R组幼鼠在I-R 6 h后肺组织病理学损伤加重,肺组织细胞质中HMGB1表达增多,血清HMGB1及BALF中IL-1β、TNF-α水平升高,肺组织MDA浓度升高,SOD活性降低,肺组织PI3K、Akt和mTOR蛋白表达上调(P<0.05);与I-R组相比,S组上述各指标均改善(P<0.05)。结论 造模前经尾静脉注射艾司氯胺酮可明显降低肝脏I-R模型幼鼠血清和肺组织中HMGB1水平,下调PI3K、Akt和mTOR蛋白表达,抑制IL-1β和TNF-α的释放,减轻氧化应激反应,从而发挥肺保护作用。

Objective To investigate the protective effect and underlying mechanism of esketamine on the lung injury induced by hepatic ischemia-reperfusion(I-R) in young mice models.Methods Eighteen C57 young mice were randomly divided into three groups with 6 mice each.Group C was taken as sham operation.The hepatic I-R models were established by hepatic ischemia for 6 hours in groups of I-R and S.Group I-R was taken as the model control.Esketamine 10 mg/kg was injected via tail vein before hepatic I-R in group S.At the time of 6 hours after hepatic I-R in groups of I-R and S and 6 hours after abdominal clearance in group C,serum, bronchoalveolar lavage fluid(BALF) and lung tissues were collected.The pathological changes of lung tissues were observed by HE staining.The levels of serum high mobility group box-1 protein(HMGB1) and TNF-α and IL-1β in BALF were determined by ELISA.The concentration of malonaldehyde(MDA) and activity of superoxide dismutase(SOD) in the lung tissues were measured by thiobarbituric acid and hydroxylamine methods, respectively.The expression of HMGB1 in the lung tissues was analyzed by immunohistochemical staining.The protein expressions of PI3K,Akt and mammalian target of rapamycin(mTOR) in the lung tissues were dectected by Western blot.Results Compared with group C,the pathological injury of lung tissues was aggravated 6 hours after I-R,the expression of HMGB1 in the cytoplasm of lung tissues was increased, the levels of serum HMGB1 and TNF-α and IL-1β in BALF were elevated, the concentration of MDA was increased, the activity of SOD was decreased, and the protein expressions of PI3K,Akt and mTOR in the lung tissues were upregulated in group I-R(P<0.05).Compared with I-R group, all the above indicators in group S were improved(P<0.05).Conclusion The injection of esketamine before model establishment can obviously decrease the level of HMGB1 in the serum and lung tissues, downregulate the protein expressions of PI3K,Akt and mTOR,inhibit the release of IL-1β and TNF-α,alleviate oxidative stress reactions, which produces a protective effect on the lung injury in young mice I-R models.