摘要
[目的]探讨槲皮素(quercetin,QUE)对脂多糖(Lipopolyaccharide,LPS)诱导的小鼠急性肺损伤(ALI)的影响及其机制。[方法]将60只雄性C57BL/6小鼠随机分为4组(每组15只):正常对照组(CON组)、QUE单独处理组(QUE组)、LPS模型组(LPS组)和QUE治疗组(LPS+QUE组)。采用腹腔注射LPS(10 mg/kg)构建小鼠ALI模型;CON组及QUE组给予等量生理盐水。QUE组只预先接受灌胃法给予QUE(25 mg/kg);LPS+QUE组在给予LPS之前2h预先灌胃QUE 25 mg/kg;CON组和LPS组灌胃等量二甲基亚碸(dimethylsulfoxide,DMSO)。采用HE染色检测肺组织病理学改变,蛋白质免疫印迹检测肺组织双链RNA依赖的蛋白激酶ER激酶(PERK)和细胞凋亡途径关键蛋白的表达。通过检测肺组织丙二醛(MDA)/谷胱甘肽(GSH)改变反映氧化应激水平,计数支气管肺泡灌洗液(BALF)中炎症细胞、炎症因子[肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和IL-1β]及肺组织中髓过氧化物(MPO)活性反映炎症程度。[结果]QUE单独处理(QUE组)对小鼠肺组织的组织病理学和炎症指标无影响。腹腔注射LPS可诱导小鼠肺组织病理损伤,增加BALF中细胞总数和中性粒细胞的数量,升高MPO活性,同时增加BALF中TNF-α、IL-6和IL-1β等炎症因子含量。而给予QUE预处理(LPS+QUE组),可显著性减轻上述改变;WB检测结果发现,QUE预处理可抑制内质网应激PERK信号途径相关蛋白的表达,减轻细胞凋亡途径关键蛋白caspase3的表达。[结论]槲皮素可通过抑制内质网应激、氧化应激及炎症反应,减轻LPS诱导的小鼠ALI,其机制与抑制内质网应激PERK信号途径有关。
[Objective]To investigate the effect of quercetin(QUE)on acute lung injury(ALI)induced by li-popolysaccharide(LPS)in mice and study its mechanism.[Methods]Sixty male C57b1/6 mice were randomly divided into 4 groups(15 in each group):normal group(CON group),LPS group and QUE treatment group.The ALI mice were constructed by intraperitoneal injection of LPS(10 mg/kg).The CON group was given the same a-mount of normal saline»while the QUE treatment group were given QUE 25 mg/kg 2 hours before the administration of LPS.The mice in both CON group and The LPS group were given the same amount of DMSO.HE staining was used to detect lung histopathological changes.Western blotting was used to detect the expression of PERK and key proteins in the apoptosis pathway.Lung tissue oxidative stress levels were detected with malondial-dehyde(MDA).Counted Inflammatory cells in bronchoalveolar lavage fluid(BLAF)and myeloperoxidase(MPO)activity in lung tissue were used to detect with inflammatory response.[Results]Quercetin alone had no effect on the morphology and inflammation of lung tissue in mice.Intraperitoneal injection of LPS induced pathological damage of mouse lung tissue which increased the number of total cells and neutrophils in BALF,increased MPO activity,and increased the levels of TNF-α and IL-1β in lung tissue.However,the QUE prevention group and the QUE treatment group significantly alleviated the above changes.Western blotting results showed that QUE pretreatment inhibited the expression of proteins related to the endoplasmic reticulum stress PERK(double-stranded RNA-dependent protein kinase ER kinase)signaling pathway and reduced the expression of caspase-3,a key protein in the apoptosis pathway.[Conclusion]Quercetin can reduce ALI in mice induced by LPS by inhibiting endoplasmic reticulum stress(ERS)and reducing oxidative stress and inflammation.The mechanism is related to the inhibition of PERK signaling pathway of ERS.
作者
马丽
肖敏
务军
冯鹏玖
董良
黄一丹
MA Li;XIAO Min;WU Jun(Department of Anesthesiology,Liuzhou Municipal People's Hospital Liuzhou Guangxi 545006)
出处
《医学临床研究》
CAS
2021年第5期647-652,共6页
Journal of Clinical Research
基金
国家自然科学基金资助项目(编号81760019)
广西自然科学基金资助项目(任务书编号:2020GXNSFAA297101)。