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,Hubei,438000,China)

机构地区：[1]黄冈市中心医院耳鼻咽喉头颈外科,湖北黄冈438000

出　　处：《中国耳鼻咽喉头颈外科》2023年第9期574-578,共5页Chinese Archives of Otolaryngology-Head and Neck Surgery

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)中调节性T细胞(regulatory T cells,Treg)/辅助性T细胞17(helper T cells17,Th17)的调节作用及潜在机制。方法 收集AR患者鼻黏膜组织及外周血单个核细胞(peripheralbloodmononuclear cell,PBMC)。饲养BALB/c小鼠。PBMC用于建立AR细胞模型,BALB/c小鼠用于建立AR动物模型。对于AR细胞使用150μg/ml的齐墩果酸进行治疗。对于AR小鼠使用低剂量(L)-齐墩果酸(50 mg/kg)、高剂量(H)-齐墩果酸(200 mg/kg)、自噬抑制剂3-MA(10 mg/kg)、H-齐墩果酸+自噬激活剂雷帕霉素(Rapa,1 mg/kg)分别进行治疗。qRT-PCR测人鼻黏膜组织IL-17A和Foxp3的表达以及小鼠各组鼻黏膜组织中RORγt和Foxp3的表达。流式细胞术从PBMC中分选Th17和Treg细胞,并检测PBMC中白细胞介素17A(IL-17A)细胞和叉头样转录因子3(Foxp3)细胞的比例。Western blot法检测Th17和Treg细胞中自噬相关蛋白Beclin1、LC3-Ⅱ、LC3-Ⅰ、p62的表达。酶联免疫吸附试验检测各组小鼠鼻腔灌洗液中IL-17A和IL-10的表达。结果 AR组患者鼻黏膜组织中IL-17A表达显著增加,但Foxp3的表达显著降低(P均<0.05)。齐墩果酸下调PBMC中IL-17A的表达,并上调Foxp3的表达。齐墩果酸抑制Th17细胞和Treg细胞中Beclin1、LC3-Ⅱ以及LC3-Ⅰ的表达(P均<0.05),促进p62的表达水平。齐墩果酸降低AR小鼠体内IL-17A和RORγt水平,并上调IL-10和Foxp3的水平(P均<0.05)。结论 齐墩果酸通过抑制自噬信号改善AR中Th17/Treg细胞的平衡。OBJECTIVE To investigate the regulatory effect of oleanolic acid on regulatory T cells(Treg)/T helper cells 17(Th17)in allergic rhinitis(AR)and its potential mechanism.METHODS Nasal mucosa tissue and peripheral blood mononuclear cell(PBMC)of AR patients were collected.BALB/c mice were raised.PBMC was used to establish AR cell models and BALB/c mice were used to establish AR animal models.AR cells were treated with oleanolic acid of 150μg/ml.AR mice were treated with low dose(L)-oleanolic acid(50 mg/kg),high dose(H)-oleanolic acid(200 mg/kg),autophagy inhibitor 3-MA(10 mg/kg),and H-oleanolic acid+autophagy activator Rapa(1 mg/kg),respectively.The expression of IL-17A and Foxp3 in human nasal mucosa and the expression of RORγt and Foxp3 in mouse nasal mucosa were measured by qRT-PCR.Th17 and Treg cells were separated from PBMC by flow cytometry,and the ratio of interleukin-17A(IL-17A)cells to fork-head transcription factor 3(Foxp3)cells in PBMC was determined.The expressions of autophagy related proteins Beclin1,LC3-II,LC3-I and p62 in Th17 and Treg cells were detected by Western blot.The expressions of IL-17A and IL-10 in nasal lavage solution were detected by enzymed-linked immunosorbent assay.RESULTS Oleanolic acid down-regulates the expression of IL-17A and up-regulates the expression of Foxp3 in PBMC(all P<0.05).Oleanolic acid inhibited the expression of Beclin1,LC3-II and LC3-I in Th17 cells and Treg cells(all P<0.05),and promoted the expression level of p62(P<0.05).Oleanolic acid decreased IL-17A and RORyt levels,and up-regulated IL-10 and Foxp3 levels in AR mice(all P<0.05).CONCLUSION Oleanolic acid can improve the balance of Th17/Treg cells in AR by inhibiting autophagy signaling.

关 键 词：齐墩果酸 鼻炎 过敏 自噬 T淋巴细胞 调节性 T淋巴细胞 辅助诱导 

分 类 号：R765.21[医药卫生—耳鼻咽喉科]