基于HMGB1/TLR4/NF-κB通路研究鱼油延缓气道重塑治疗哮喘的作用机制  

Based on HMGB1/TLR4/NF-κB Signaling Pathway Study on the Mechanism of Fish Oil Delaying Airway Remodeling in the Treatment of Asthma

作　　者：曹娟[1] 赵辉[2] 牛忻 李素霞 于淼[2] 袁莉[2] CAO Juan;ZHAO Hui;NIU Xin(Department of Traditional Chinese Medicine,Heze Medical College,Heze City,Shandong Province 274000;不详)

机构地区：[1]山东省菏泽医学专科学校中医药系,山东省菏泽市274000 [2]山东省菏泽医学专科学校基础医学部,山东省菏泽市274000

出　　处：《医学理论与实践》2024年第10期1621-1626,共6页The Journal of Medical Theory and Practice

基　　金：山东省医药卫生科技发展计划项目(2019WS342)。

摘　　要：目的:探讨鱼油通过HMGB1/TLR4/NF-κB信号通路延缓气道重塑,治疗哮喘的潜在作用机制及其疗效,为哮喘的新型治疗策略提供理论依据和实验支持。方法:采用Ovalbumin(OVA)诱导的哮喘小鼠模型,随机分为对照组、模型组、鱼油组、鱼油+E5564组,每组10只,共40只;4组小鼠,通过肺功能测试仪对比气道阻力,酶联免疫吸附试验判断运用TLR4抑制剂前后的IL-1β、IL-6、TNF-α、IL-10炎症因子水平,苏木精—伊红染色观察肺组织病理变化,逆转录定量聚合酶链反应对比Bax、caspase-3、Bcl2的mRNA表达水平,免疫印迹分析观察Bax、caspase-3、Active-caspase3、HMGB1、TLR4、NF-κB、p-NF-κB p65的蛋白表达量,免疫组化法观察HMGB1、TLR4、NF-κB的蛋白表达水平。结果:气道阻力测试表明,鱼油组小鼠的气道阻力显著降低,肺组织病理损伤减轻,炎症细胞浸润明显减少;HE染色结果表明降低,鱼油组小鼠的气道损伤更小,炎症细胞浸润和上皮细胞排列紊乱更少;ELISA结果显示,鱼油组小鼠的血清炎症因子IL-1β、IL-6、TNF-α的表达降低,并提高了IL-10的表达,而鱼油+E5564组的血清炎症因子水平与模型组相近;RT-qPCR结果表明,鱼油组小鼠的Bax、caspase-3的mRNA表达量较低,Bcl2的mRNA表达量较高;WB结果表明,鱼油组小鼠的Bax、caspase-3、Active-caspase3、HMGB1、TLR4、NF-κB、p-NF-κB p65蛋白表达量更低,而鱼油+E5564组小鼠HMGB1、TLR4、NF-κB及p-NF-κB p65蛋白表达量与模型组小鼠相近;IHC结果表明,鱼油组小鼠HMGB1、TLR4、NF-κB蛋白表达量更低,与WB结果趋同。结论:鱼油通过影响HMGB1/TLR4/NF-κB信号通路,显著减轻哮喘小鼠模型的气道炎症和重塑,提高肺功能,显示出作为哮喘潜在治疗剂的可能性;该研究为鱼油在哮喘治疗领域的应用提供了实验依据,但需要进一步的临床研究来验证其疗效和安全性。Objective:To investigate the potential mechanisms and therapeutic efficacy of fish oil in delaying airway remodeling and treating asthma through the HMGB1/TLR4/NF-κB signaling pathway,providing theoretical and experimental support for novel asthma treatment strategies.Methods:An asthma mouse model was induced using Ovalbumin(OVA)and randomly divided into four groups:control,model,fish oil,and fish oil+E5564,with 10 mice in each group for a total of 40.The airway resistance was compared using a lung function tester,the levels of inflammatory cytokines IL-1β,IL-6,TNF-α,and IL-10 were measured using ELISA before and after the application of a TLR4 inhibitor,lung tissue pathological changes were observed through Hematoxylin and Eosin staining,mRNA expression levels of Bax,caspase-3,and Bcl2 were compared using RT-qPCR,and the protein expression levels of Bax,caspase-3,Active-caspase3,HMGB1,TLR4,NF-κB,and p-NF-κB p65 were observed through Western blot analysis.Immunohistochemistry was used to observe the protein expression levels of HMGB1,TLR4,and NF-κB.Results:Airway resistance tests showed that fish oil significantly reduced airway resistance in mice,decreased lung tissue damage,and lessened inflammatory cell infiltration.HE staining indicated that fish oil mice had less airway damage and reduced inflammatory cell infiltration and epithelial cell disarray.ELISA results showed reduced serum inflammatory cytokine levels of IL-1β,IL-6,TNF-α,and increased IL-10 expression in fish oil mice,while the fish oil+E5564 group had similar cytokine levels to the model group.RT-qPCR results indicated lower mRNA expression levels of Bax and caspase-3,and higher Bcl2 expression in fish oil mice.Western blot analysis showed lower protein expression levels of Bax,caspase-3,Active-caspase3,HMGB1,TLR4,NF-κB,and p-NF-κB p65 in fish oil mice,with the fish oil+E5564 group showing similar levels to the model group.Immunohistochemistry results were consistent with Western blot findings,showing lower expression levels of HMGB1,TLR4

关 键 词：哮喘 HMGB1/TLR4/NF-κB 信号通路 鱼油 动物实验 气道炎 

分 类 号：R-33[医药卫生]