摘要
目的:探讨白藜芦醇(RES)调控miR-125b-5p/PRDM1轴在改善儿童哮喘气道炎症反应中的作用及其分子机制。方法:采用卵清蛋白(OVA)诱导哮喘大鼠模型。将大鼠随机分为假手术组(sham)、哮喘组(model)、10μmol/L RES(RES-L)组、50μmol/L RES(RES-H)组、50μmol/L miR-125b-5p antagomir(miR-125b-5p antagomir)组和miR-125b-5p antagomir+50μmol/L RES(miR-125b-5p antagomir+RES)组。检测大鼠支气管肺泡灌洗液(BALF)中炎症细胞数量。通过HE染色及Masson三色染色观察肺组织的病理改变。通过ELISA检测肺匀浆中炎症因子水平。并采集哮喘儿童和健康儿童的血清进行分析。采用RT-qPCR检测哮喘和健康儿童以及不同组大鼠肺组织中miR-125b-5p、PRDM1的表达。双荧光素酶实验验证miR-125b-5p与PRDM1的靶向结合关系。结果:HE染色结果表明,哮喘组大鼠气道上皮发生了多种炎症细胞浸润,而RES组的病理改变有所减轻。Masson染色表明,与哮喘组相比,RES组气道胶原沉积面积减少。哮喘组大鼠BALF中炎症细胞数量高于RES组。RES治疗显著降低了气道壁和平滑肌的厚度。ELISA结果显示,哮喘组肺组织中IL-1β、IL-10和TNF-α水平升高,IL-12水平降低,而RES组则表现出相反的趋势。哮喘组大鼠肺组织中miR-125b-5p水平显著升高、PRDM1水平显著降低,RES治疗可逆转这一结果。此外,miR-125b-5p antagomir治疗可减轻哮喘大鼠的气道炎症和气道重塑。双荧光素酶实验证实miR-125b-5p与PRDM1存在靶向结合关系。与健康儿童相比,哮喘儿童血清IL-1β、IL-10和TNF-α水平升高,IL-12水平降低。结论:RES通过HMGB1/TLR4/NF-κB途径抑制炎症细胞因子释放,减轻了哮喘诱导的气道炎症和气道重塑。
Objective:To investigate the role of resveratrol(RES)in improving airway inflammation in children with asthma by regulating miR-125b-5p/PRDM1 axis and its molecular mechanism.Methods:Rat model of asthma was induced by ovalbumin(OVA).The rats were randomly divided into sham group(sham),asthma group(model),10μmol/L RES(RES-L)group,50μmol/L RES(RES-H)group,50μmol/L miR-125b-5p antagomir(miR-125b-5p antagomir)group and miR-125b-5p antagomir+50μmol/L RES(miR-125b-5p antagomir+RES)group.The number of inflammatory cells in bronchoalveolar lavage fluid(BALF)was detected.HE staining and Masson three-color staining were used to observe pathological changes of lung tissue.Levels of inflammatory factors in lung homogenates were detected by ELISA.Serum from asthmatic children and healthy children was collected for analysis.Expressions of miR-125b-5p and PRDM1 in lung tissues of asthmatic and healthy children and different groups of rats were detected by RT-qPCR.Targeted binding relationship between miR-125b-5p and PRDM1 was detected by double luciferase assay.Results:HE staining showed that the airway epithelium in asthma group had a variety of inflammatory cell infiltration,while pathological changes in RES group were reduced.Masson staining showed that the area of airway collagen deposition was reduced in RES group compared with asth-ma group.The number of inflammatory cells in BALF in asthma group was higher than that in RES group.RES treatment significantly reduced airway wall and smooth muscle thickness.ELISA results showed that lung tissue in the asthma group had increased levels of IL-1β,IL-10 and TNF-α,while decreased level of IL-12,the RES group showed the opposite trend.Level of miR-125b-5p in lung tissue of asthmatic rats was significantly increased,while level of PRDM1 was decreased,and RES treatment could reverse this result.In addi-tion,treatment with miR-125b-5p antagomir alleviated airway inflammation and airway remodeling in asthmic rats.Dual luciferase assay confirmed the targeting binding relationship between miR-125b-5p and PRDM1.Compared with healthy children,serum levels of IL-1β,IL-10 and TNF-αwere increased,while IL-12 level was decreased in children with asthma.Conclusion:RES inhibits the re-lease of inflammatory cytokines through HMGB1/TLR4/NF-κB pathway,alleviating asthma-induced airway inflammation and airway remodeling.
作者
苏丹
刘亚楠
张琳
SU Dan;LIU Yanan;ZHANG Lin(Hengshui People's Hospital,Hengshui 053000,China;Hengshui Maternal and Child Health Hospital,Hengshui 053000,China)
出处
《中国免疫学杂志》
北大核心
2025年第8期1901-1907,共7页
Chinese Journal of Immunology
基金
河北省中医药类科学研究课题计划项目(2023462)。
