橙花叔醇对脓毒症急性肺损伤大鼠炎症反应及AMPK/SIRT1通路的影响

Effect of Nerolidol on the Inflammatory Response and AMPK/SIRT1Pathway in Rats with Sepsis-induced Acute Lung Injury

【目的】探讨橙花叔醇对脓毒症急性肺损伤大鼠的治疗作用及机制。【方法】采用脂多糖(LPS)股静脉注射法构建脓毒症急性肺损伤大鼠模型,应用橙花叔醇进行预防性治疗。测试用力呼出25%~75%肺活量时的呼气流量(FEF25%-75%)和20毫秒用力呼气量/用力肺活量(FEV20/FVC)比值评估大鼠肺功能。收集大鼠支气管肺泡灌洗液(BALF)和肺组织,采用苏木素-伊红(HE)染色法观察肺组织病理形态,应用血细胞计数器计数BALF中总白细胞数和中性粒细胞数,采用酶联免疫吸附分析(ELISA)检测BALF中炎症因子白细胞介素(IL)-1、IL-10和肿瘤坏死因子(TNF)-α水平,采用Western Blot法检测肺组织单磷酸腺苷活化蛋白激酶(AMPK)/SIRT1通路相关蛋白表达。继而给予AMPK抑制剂Dorsomorphin进行干预,进一步观察橙花叔醇通过激活AMPK/SIRT1通路对LPS诱导的脓毒症急性肺损伤大鼠的影响。【结果】橙花叔醇可减轻LPS诱导的脓毒症急性肺损伤大鼠肺组织病理及肺功能损伤,降低BALF中总白细胞数、中性粒细胞数及炎症因子IL-1和TNF-α水平,提高BALF中IL-10水平,上调肺组织磷酸化AMPK(p-AMPK)、SIRT1蛋白表达水平。【结论】橙花叔醇可通过激活AMPK/SIRT1通路及减轻炎症反应改善脓毒症急性肺损伤大鼠。

Objective To investigate the therapeutic effect and mechanism of nerolidol on acute lung injury in rats with sepsis.Methods A rat model of acute lung injury in sepsis was constructed using lipopolysaccharide(LPS)femoral intravenous injection,and nerolidol was applied as prophylactic treatment.The forced expiratory flow at 25%to 75%of forced vital capacity(FEF25%-75%)and forced expiratory volume in 20 ms/forced vital capacity(FEV20/FVC)ratio were tested to assess the lung function of rats.The bronchoalveolar lavage fluid(BALF)and lung tissue were collected from rats,and hematoxylin-eosin(HE)staining was used to observe the histopathological morphology of lung.A blood cell counter was applied to perform the total leukocyte count and neutrophil count in BALF,and the levels of inflammatory factors interleukin(IL)-1,IL-10 and tumour necrosis factor(TNF)-αwere measured by enzyme-linked immunosorbent assay(ELISA),and lung tissue adenosine monophosphate-activated protein kinase(AMPK)/SIRT1 pathway-related protein expression was measured by Western Blot.Dorsomorphin,an AMPK inhibitor,was administered to further investigate the effect of nerolidol on LPS-induced acute lung injury in rats with sepsis through activation of the AMPK/SIRT1 pathway.Results Nerolidol attenuated lung histopathology and lung function impairment in rats with LPS-induced acute lung injury in sepsis,decreased total leukocyte count,neutrophil count and inflammatory factors IL-1 and TNF-αlevels in BALF,increased IL-10 level in BALF,and up-regulated lung tissue phosphorylated AMPK(p-AMPK)and SIRT1 protein expression levels.Conclusion Nerolidol improves acute lung injury in rats with sepsis by activating the AMPK/SIRT1 pathway and reducing the inflammatory response.