川续断皂苷乙通过调节细胞免疫平衡对哮喘小鼠气道炎症的减轻作用

Alleviating airway inflammation in asthmatic mice by regulating cellular immune balance

目的 旨在评估川续断皂苷乙(DB)对哮喘小鼠气道炎症的影响,并初步探讨潜在的机制。方法 采用卵清蛋白(OVA)构建哮喘小鼠模型。将小鼠按随机数字表法随机分为5组:正常对照组、模型(OVA)组、地塞米松(DEX)组[灌胃给予0.075 mg/(kg·d)地塞米松]、低剂量DB(L-DB)组[灌胃给予10 mg/(kg·d)川续断皂苷乙]和高剂量DB(H-DB)组[灌胃给予60 mg/(kg·d)川续断皂苷乙]。采用HE染色、Masson染色和过碘酸雪夫(PAS)染色进行组织病理学分析。通过酶联免疫吸附法(ELISA)检测支气管肺泡灌洗液(BALF)和血清中的炎症介质水平。通过流式细胞仪分析肺细胞中的T辅助细胞(Th)和调节性T细胞(Treg)的占比。结果 与正常对照组相比,OVA组小鼠血清IgE、肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)水平、BALF中干扰素-γ(IFN-γ)、IL-10水平显著降低,而IL-13、IL-4、IL-17、转化生长因子-β1(TGF-β1)水平、BALF中嗜酸性粒细胞(EOS)、淋巴细胞(LYMs)、中性粒细胞(NEU)和巨噬细胞(MAC)的数量、Th2细胞亚群和Th17细胞亚群占比显著升高,而白细胞介素-10(IL-10)水平、BALF中干扰素-γ(IFN-γ)、IL-10水平、肺组织中Th1细胞亚群和Treg细胞亚群占比显著降低(P均<0.01);与OVA组相比,DEX组和H-DB组小鼠血清IgE、TNF-α和IL-6水平、BALF中IL-13、IL-4、IL-17、TGF-β1水平、BALF中EOS、LYMs、NEU和MAC的数量、肺组织Th2细胞亚群和Th17细胞亚群占比显著降低,而IL-10水平、BALF中IFN-γ、IL-10水平、肺组织中Th1细胞亚群和Treg细胞亚群占比显著升高(P均<0.01)。结论 DB能有效改善哮喘小鼠的症状、肺组织损伤及气道炎症反应,这可能是通过调节Th1/Th2和Th17/Treg细胞免疫平衡来实现。

Objective The aim of this study was to evaluate the effects of Dipsacoside B(DB)on airway inflammation in asthmatic mice and to preliminarily explore the potential mechanisms.Methods An asthma mouse model was constructed using ovalbumin(OVA).The mice were randomly divided into five groups according to the random number table method:normal control group,model(OVA)group,dexamethasone(DEX)group(0.075 mg/(kg·d)dexamethasone was given by gavage,low-dose DB(L-DB)group(10 mg/(kg·d)Chuanxuijian saponin ethyl was given by gavage),and high dose DB(H-DB)group(60 mg/(kg·d))Chuanxuijian saponin B).Histopathological analyses were performed using hematoxylin eosin staining(HE),Masson staining and peroxynitrite Schiff(PAS)staining.Inflammatory mediator levels in bronchoalveolar lavage fluid(BALF)and serum were detected by enzyme-linked immunosorbent assay(ELISA).The percentage of T helper(Th)and regulatory T cells(Treg)in lung cells was analysed by flow cytometry.Results Serum levels of IgE,tumour necrosis factor-α(TNF-α)and interleukin-6(IL-6)were significantly high-er,while interleukin-10(IL-10)level was significantly lower in the OVA group of mice compared to that in the normal control group(P<0.05).Serum IgE,TNF-αand IL-6 levels were significantly lower,while IL-10 level was significantly higher(P<0.05)in DEX and H-DB groups of mice compared to OVA group.Compared with the normal control group,the levels of interferon-γ(IFN-γ)and IL-10 were significantly lower,while the levels of interleukin-13(IL-13),interleukin-4(IL-4),interleukin-17(IL-17),and transforming growth factor-β1(TGF-β1)were significantly higher in the BALF of mice in the OVA group,compared with that in the OVA group(P<0.05).The levels of IFN-γand IL-10 were significantly higher,while the levels of IL-13,IL-4,IL-17,and TGF-β1 were signifi-cantly lower in the BALF of mice in the DEX and H-DB groups(P<0.05).The numbers of eosinophils(EOS),lymphocytes(LYMs),neutrophils(NEU),and macrophages(MAC)were significantly increased in BALF of the model mice compared to that in the normal control group(P<0.05).The numbers of EOS,LYMs,NEU and MAC were significantly lower in the BALF of mice in the DEX and H-DB groups compared to that in the OVA group(P<0.05).Compared with the normal control group,the percentages of Th1 cell sub-population and Treg cell subpopulation in the lung tissues of mice in the OVA group were significantly lower,while the percentage of Th2 cell subpopulation and Th17 cell subpopulation was significantly higher(P<0.05).Compared with the OVA group,the percenta-ges of Th1 cell subpopulation and Treg cell subpopulation in the lung tissues of mice in the DEX group and the H-DB group were signif-icantly higher,while the percentages of Th2 cell subpopulation and Th17 cell subpopulation were significantly lower(P<0.05).Conclusion DB can effectively remit the symptoms,lung tissue damage and airway inflammatory response in asthmatic mice,which may be achieved by regulating the immune balance of Th1/Th2 and Th17/Treg cells.