白藜芦醇通过ERK1/2和NF-κB通路调节NADPH氧化酶表达改善急性肺损伤

Resveratrol regulates NADPH oxidase expression through ERK1/2 and NF-κB pathways to improve acute lung injury

目的探讨白藜芦醇对急性肺损伤大鼠NADPH氧化酶的影响,并分析相关信号通路机制。方法将50只雄性SPF级SD大鼠随机分为对照组、模型组、地塞米松组、低剂量白藜芦醇组和高剂量白藜芦醇组,每组10只。对照组和模型组给予生理盐水干预,地塞米松组、低剂量白藜芦醇组和高剂量白藜芦醇组分别给予2 mg/kg地塞米松、100 mg/kg白藜芦醇、500 mg/kg白藜芦醇干预,1次/d,连续干预14 d。干预结束后,除对照组外,其余组别大鼠构建急性肺损伤模型。比较各组大鼠肺组织病理状态、肺湿重/干重比(W/D)、肺损伤评分,采用ELISA法检测肺泡灌洗液白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)水平;试剂盒检测丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)水平;采用RT-PCR法检测各组NADPH氧化酶亚基p47^(phox)、p22^(phox),及ERK、NF-κB mRNA水平;采用Western bloting法检测各组p47^(phox)、p22^(phox)、p-ERK1/2、ERK1/2、p-NF-κB、NF-κB蛋白水平。结果模型组、地塞米松组、低剂量白藜芦醇组和高剂量白藜芦醇组W/D及肺损伤评分、MDA水平、p22^(phox)、p47^(phox)、ERK1/2、NF-κB mRNA、p47^(phox)蛋白、p-ERK/ERK、p-NF-κB/NF-κB、IL-6、IL-8、TNF-α水平高于对照组,而SOD、GSH-Px水平低于对照组(P<0.05)。结论白藜芦醇可改善LPS诱导的急性肺损伤大鼠NADPH氧化酶及氧化应激状态,其作用机制可能与ERK1/2-NF-κB通路有关。

Objective To investigate the effect of resveratrol on NADPH oxidase in rats with acute lung injury and to analyze the related signaling pathway mechanism.Methods Fifty male SD rats were randomly divided into control group,model group,dexamethasone group,low-dose group and high-dose group,with 10 rats in each group.The control group and model group were given normal saline intervention,and the dexamethasone group,low-dose resveratrol group and high-dose resveratrol group were respectively given 2 mg/kg dexamethasone,100 mg/kg resveratrol and 500 mg/kg resveratrol intervention once a day for 14 days.After the intervention was completed,acute lung injury models were constructed for the rats of the remaining groups,except the control group.The pathological status of lung tissue,the ratio of lung wet/dry weight(W/D)and lung injury score were compared in each group.The levels of interleukin-6(IL-6),interleukin-8(IL-8)and tumor necrosis factorα(TNF-α)in alveolar lavage fluid were detected by ELISA.The levels of malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)were detected with the kit.The mRNA levels of NADPH oxidase subunits p47^(phox),p 22phox,ERK and NF-κB were detected by RT-PCR.The protein levels of p47^(phox),p22^(phox),p-ERK1/2,ERK1/2,p-NF-κB and NF-κB in each group were detected by Western blot.Results W/D,lung injury score,MDA level,P22^(phox),P47^(phox),ERK1/2,NF-κB mRNA,p22^(phox),p47^(phox) protein,p-ERK/ERK,p-NF-κB/NF-κB,IL-6,IL-8,TNF-αlevels in model group,dexamethasone group,low-dose resveratrol group and high-dose resveratrol group were higher than those in control group,while the SOD and GSH-Px levels were lower than control group(P<0.05).Conclusion Resveratrol can improve NADPH oxidase and oxidative stress in LPS-induced acute lung injury rats,the mechanism of which may be related to ERK1/2-NF-κB pathway.