芒柄花黄素通过DRP1-NLRP3信号通路缓解过敏性哮喘的作用机制

Mechanism of action of formononetin in alleviating allergic asthma through DRP1-NLRP3 signaling pathway

目的本研究旨在探讨芒柄花黄素(formononetin,FN)通过干预ROS的生成抑制线粒体动力相关蛋白1(dynamic-related protein 1,DRP1)-NLRP3轴减轻过敏性气道炎症的有关机制。方法为建立过敏性哮喘小鼠模型,50只8周龄的BALB/c小鼠通过卵清蛋白(ovalbumin,OVA)诱导后分为对照组、模型组、FN治疗组及地塞米松组。采用HE和Masson染色检测气道炎症和胶原沉积,ELISA测定支气管肺泡灌洗液(BALF)中Th2型细胞因子和超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA)以及IgE水平,DCFH-DA染色评估BEAS-2B细胞中ROS,免疫组织化学和免疫荧光检测肺组织和BEAS-2B细胞中DRP1表达,免疫印迹分析DRP1-NLRP3途径。结果FN治疗可有效改善哮喘小鼠模型的症状,包括减少嗜酸性粒细胞聚集、气道胶原沉积,降低Th2细胞因子和IgE水平,减少ROS和MDA生成,提高SOD和CAT活性,并调节DRP1-NLRP3途径相关蛋白表达,从而缓解炎症。结论FN通过调节DRP1-NLRP3途径改善哮喘气道炎症。

Aim To investigate the mechanism by which formononetin(FN)inhibits mitochondrial dynamic-related protein 1(DRP1)-NLRP3 axis via intervening the generation of ROS to reduce allergic airway inflammation.Methods In order to establish allergic asthma mouse model,50 BALB/c mice aged 8 weeks were divided into the control group,model group,FN treatment group and dexamethasone group after ovalbumin(OVA)induction.Airway inflammation and collagen deposition were detected by HampE and Masson staining.Th2 cytokines and superoxide dismutase(SOD),catalase(CAT),malondialdehyde(MDA),and IgE levels in bronchoalveolar lavage fluid(BALF)were measured by ELISA,ROS in BEAS-2B cells was assessed by DCFH-DA staining,DRP1 expression in lung tissue and BEAS-2B cells was detected by immunohistochemistry and immunofluorescence,and the DRP1-NLRP3 pathway was analyzed by immunoblotting.Results FN treatment could effectively ameliorate the symptoms of asthmatic mouse model,including reducing eosinophil accumulation,airway collagen deposition,decreasing Th2 cytokine and IgE levels,reducing ROS and MDA production,increasing SOD and CAT activities,and regulating DRP1-NLRP3 pathway-related protein expression,thereby relieving inflammation.Conclusion FN ameliorates airway inflammation in asthma by regulating DRP1-NLRP3 pathway.