白藜芦醇对中性粒细胞性哮喘小鼠肺组织炎症的抑制作用及其机制

Inhibitory effect of resveratrol on pulmonary inflammation in mice with neutrophilic asthma and its mechanism

目的:探讨白芦藜醇对中性粒细胞性哮喘小鼠肺组织炎症的抑制作用,并阐明其作用机制。方法:选取18只C57BL/6J雌性小鼠,采用脂多糖(LPS)+卵白蛋白(OVA)联合致敏方法建立中性粒细胞性哮喘小鼠模型,随机分为模型组(LPS+OVA组)、白藜芦醇组(Res组,于激发前2h灌胃30mg·kg^(-1)白藜芦醇)和N-乙酰半胱氨酸组(NAC组,于激发前2h灌胃3mmol·kg^(-1) N-乙酰半胱氨酸);同时选取6只同周龄小鼠作为正常对照组(PBS组)。于乙酰甲胆碱雾化期间观察各组小鼠的行为学变化,采用肺功能仪分析小鼠气道高反应(AHR),光学显微镜观察支气管肺泡灌洗液(BALF)中细胞总数和炎症细胞,HE染色观察各组小鼠肺组织病理形态表现,酶联免疫法(ELISA)检测各组小鼠BALF上清中白细胞介素17(IL^(-1)7)水平,特异性荧光探针DCF-DA染色分析肺组织活性氧(ROS)水平,丙二醛(MDA)试剂盒检测肺组织匀浆中MDA水平。结果:与PBS组比较,LPS+OVA组小鼠Penh值、细胞总数、炎症细胞、气道炎症及评分均明显升高(P<0.05或P<0.01),BALF上清中IL^(-1)7水平明显升高(P<0.01),肺组织内ROS荧光强度明显升高,肺匀浆中MDA水平明显升高(P<0.01)。与LPS+OVA组比较,Res组和NAC组小鼠Penh值、细胞总数、炎症细胞、气道炎症及评分明显降低(P<0.05或P<0.01),BALF上清中IL^(-1)7水平均明显降低(P<0.05),肺组织内ROS荧光强度降低,肺匀浆中MDA水平明显降低(P<0.01)。结论:白藜芦醇能够有效抑制中性粒细胞性哮喘小鼠肺组织中的炎症反应,其机制可能与白藜芦醇抑制体内发生的氧化应激反应有关。

Objective:To investigate the inhibitory effect of resveratrol on the pulmonary inflammation in the mice with neutrophilic asthma,and to elucidate its mechanism.Methods:A total of 18 female C57BL/6J mice were selected.The mouse models of neutrophilic asthma were established by sensitization with lipopolysaccharide(LPS)and ovalbumin(OVA).The model mice were randomly divided into model group(LPS+OVA group),resveratrol group(Res group,intragastric administration of 30 mg·kg-1 resveratrol at 2 h before challenge)and N-acetylcysteine group(NAC group,intragastric administration of 3 mmol·kg-1 N-acetylcysteine at 2 h before challenge).Other 6 age-matched female C57BL/6J mice were choosen as normal control group(PBS group).The behavioral changes of mice in various groups were observed during the atomization with methacholine;the airway hyperresponsiveness(AHR)of mice was analyzed with a lung function instrument,and the total number of cells and inflammatory cells in the bronchoalveolar lavage fluid(BALF)were observed with light microscope;the pathological changes of lung tissue were observe by HE staining;the levels of interleukin-17(IL-17)in the supernatant of BALF of the mice in various groups were detected by enzyme-linked immunosorbent assay(ELISA);the level of reactive oxygen species(ROS)in lung tissue was analyzed by specific fluorescent probe DCF-DA staining;the level of malondialdehyde(MDA)in lung homogenate was measured by MDA kit.Results:Compared with PBS group,the Penh value,total number of cells,inflammatory cells,airway inflammation and scores of the mice in LPS+OVA group were significantly increased(P<0.05 or P<0.01);the IL-17 level in BALF supernatant was significantly increased(P<0.01);the ROS fluorescence intensity in lung tissue was significantly increased,and the MDA level in lung homogenates was significantly increased(P<0.01).Compared with LPS+OVA group,the Penh value,total number of cells,inflammatory cells,airway inflammation and scores in Res group and NAC group were significantly decreased(P<0.05 or P<0.01);the IL-17 levels in BALF supernatant were significantly decreased(P<0.05 or P<0.01);the ROS fluorescence intensities in lung tissue were decreased,and the MDA levels in lung homogenates were significantly decreased(P<0.01).Conclusion:Resveratrol can effectively inhibit the inflammatory response of lung tissue in the mice with neutrophilic asthma,and its mechanism may be related to inhibiting the oxidative stress reaction.