小檗碱通过抑制Th1恢复IFN-γ/IL-4平衡减轻迟发型超敏反应

Berberine Attenuates Delayed Type Hypersensitivity by Inhibiting Th1 and Restoring IFN-γ/IL-4 Balance

目的:观察小檗碱对卵清蛋白(ovalbumin,OVA)诱导的迟发型超敏反应(delayed type hypersensitivity,DTH)模型小鼠的作用并探讨其机制。方法:32只小鼠随机分为对照组、DTH组、小檗碱组、DTH+小檗碱组,制备DTH模型给予小檗碱处理;测量小鼠足垫肿胀厚度,苏木精-伊红染色(hematoxylin-eosin staining,HE)法观察炎性细胞浸润,酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)检测足垫组织匀浆中γ干扰素(interferon-γ,IFN-γ)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白介素-4(interleukin-4,IL-4)的表达,制备脾脏单个核细胞,刀豆蛋白(concanavalin A,Con A)实验观察IFN-γ水平,卵清蛋白(ovalbumin,OVA)致敏实验检测上清IFN-γ、IL-4表达。结果:小檗碱处理明显降低小鼠足垫肿胀程度,减少炎症细胞浸润,抑制足垫组织中IFN-γ、TNF-α炎性因子释放,并减少脾脏组织中IFN-γ的分泌,抑制Th1反应,恢复IFN-γ/IL-4平衡(P<0.01,P<0.001),差异均有统计学意义。结论:小檗碱通过抑制Th1反应,恢复IFN-γ/IL-4平衡发挥对迟发型超敏反应的抑制作用。

Objective:To investigate the role of berberine on mice with ovalbumin(OVA)-induced delayed hypersensitivity(DTH)model and explore its possible mechanisms.Methods:Thirty-two mice were randomly divided into control group,DTH group,berberine group and DTH+berberine group.Firstly,DTH model was prepared and mice were treated with berberine.The thickness of mouse footpad was measured;inflammatory cell infiltration was detected by HE staining;IFN-γ,TNF-αand IL-4 levels were detected by ELISA from homogenization of footpad tissue;Spleen lymphocytes were isolated and cultured.ConA experiment was conducted to detect the secretion of IFN-γ,and OVA sensitization experiment was used to assess IFN-γand IL-4 levels.Results:Berberine treatment significantly reduced the thickness of footpad,decreased the infiltration of inflammatory cells,inhibit the expression of IFN-γand TNF-αin the footpad tissue and lowered the production of IFN-γin spleen lymphocytes.Importantly,berberine treatment inhibited Th1 response and restored IFN-γ/IL-4 balance,which was associated with the anti-inflammatory effect of berberine on OVA-induced DTH(P<0.01,P<0.001).Conclusion:Berberine alleviates delayed hypersensitivity by restoring IFN-γ/IL-4 balance via inhibiting Th1 response.