齐墩果酸通过抑制自噬调节变应性鼻炎中的Treg/Th17细胞失衡  

Oleanolic acid regulates Treg/Th17 cell imbalance in allergic rhinitis by inhibiting autophagy

作　　者：王丽[1] 黄永阳 张义 綦彬 万龙[1] 杜炎平 陈才盛[1] WANG Li;HUANG Yongyang;ZHANG Yi;QI Bin;WAN Long;DU Yanping;CHEN Caisheng(Department of Otolaryngology Head and Neck Surgery,Huanggang Central Hospital,Huanggang 438000,China)

机构地区：[1]黄冈市中心医院耳鼻喉头颈外科,黄冈438000

出　　处：《中国免疫学杂志》2024年第7期1399-1404,共6页Chinese Journal of Immunology

基　　金：湖北省卫生和计划生育委员会科研项目(WJ2017F087)。

摘　　要：目的:探讨齐墩果酸对变应性鼻炎(AR)的作用及潜在作用机制。方法:密度梯度离心分离外周血单个核细胞(PBMCs)。实时定量PCR检测鼻黏膜中Foxp3、IL-17A、Beclin1、RORγt mRNA表达;流式细胞术检测PBMCs中Treg和Th17细胞比例;Western blot检测鼻黏膜中RORγt、LC3-Ⅰ、LC3-Ⅱ、Beclin1、P62蛋白表达。将小鼠随机分为对照组、卵清蛋白(OVA)组、低剂量(L-)齐墩果酸组、高剂量(H-)齐墩果酸组、DSGC、3-MA和H-齐墩果酸+Rapa组。HE染色和高碘酸席夫(PAS)染色观察组织学变化。ELISA检测血清学变化、IL-17A、IL-10、嗜酸性阳离子蛋白(ECP)和抗OVA特异性抗体表达。结果:AR患者存在Th17/Treg失衡和自噬激活,而齐墩果酸在体外恢复Th17/Treg平衡并抑制自噬。体内实验表明,齐墩果酸可抑制OVA诱导的过敏性鼻部症状和自噬通路激活,与3-MA的调节作用相同,而Rapa可减弱高剂量齐墩果酸的作用。此外,齐墩果酸可减少鼻腔灌洗液中不同炎症细胞浸润,防止上皮细胞损伤,改善鼻黏膜厚度及黏液分泌。齐墩果酸可调节小鼠Th17/Treg相关细胞因子表达平衡。结论:齐墩果酸是治疗AR极具潜力的药物,通过抑制自噬调节Th17/Treg平衡发挥作用。Objective:To explore effect of oleanolic acid on allergic rhinitis(AR)and its potential mechanism.Methods:Peripheral blood mononuclear cells(PBMCs)were isolated by density gradient centrifugation.Real-time quantitative PCR was used to detect expressions of Foxp3,IL-17A,Beclin1,and RORγt mRNA in nasal mucosa;flow cytometry was employed to assess proportion of Treg and Th17 cells in PBMCs;Western blot was utilized to determine expressions of RORγt,LC3-Ⅰ,LC3-Ⅱ,Beclin1,and P62 proteins in nasal mucosal.Mice were randomly divided into Control,ovalbumin(OVA),L-oleanolic acid,H-oleanolic acid,DSGC,3-MA and H-oleanolic acid+Rapa groups.Histological changes were observed by HE staining and Periodate Schiff(PAS)staining.Serological changes,expressions of IL-17A,IL-10,eosinophilic cationic protein(ECP)and anti-OVA-specific antibodies were detected by ELISA.Results:AR patients had Th17/Treg imbalance and activated autophagy.Oleanolic acid restored Th17/Treg balance and inhibited autophagy in vitro.In vivo experiments demonstrated that oleanolic acid inhibited OVA-induced allergic nasal symptoms and activation of autophagy pathway,which was same as regulation of 3-MA,while Rapa could weaken effects of high dose of oleanolic acid.Moreover,oleanolic acid reduced infiltration of different inflammatory cells of nasal lavage fluid,prevented damages to epithelial cells,and improved nasal mucosal thickness and mucus secretion.Oleanolic acid regulated balance of mouse Th17/Tregassociated cytokines expressions.Conclusion:Oleanolic acid can be used to treat AR,whose effect is realized through inhibiting autophagy to regulate Th17/Treg balance.

关 键 词：齐墩果酸 变应性鼻炎 自噬 TH17/TREG 

分 类 号：R285[医药卫生—中药学]