温阳补肾方对哮喘小鼠脾脏树突细胞的免疫调控机制研究

Immunoregulatory mechanism of "Wenyang Bushen Decoction" on spleen dendrictic cells in mice with asthma

目的通过观察哮喘小鼠脾脏中树突细胞(DC)表面共刺激分子CD80、CD86的表达变化,探讨温阳补肾方对哮喘小鼠脾脏树突细胞的免疫调控机制。方法 30只BALB/c小鼠随机分为正常对照组、模型组、中药组、西药组、中西药结合组,每组6只。除正常组外,余组建立小鼠哮喘模型,并予以相应的干预措施。各组小鼠肺组织切片进行苏木精-伊红染色后观察炎症反应;采用酶联免疫吸附法(ELISA)测定血清中IL-4、IFN-γ、ECP的含量;取各组小鼠脾脏组织分离树突细胞,经过DC培养,扫描电镜观察DC形态,用流式细胞仪(FACS)分析各组小鼠脾脏DC表面共刺激分子CD80、CD86的表达量。结果模型组小鼠气道上皮损伤、脱落,嗜酸性粒细胞等炎性细胞浸润。与正常对照组比较,模型组小鼠IL-4、ECP水平上调,INF-γ水平下调,CD80、CD86阳性表达率上调,差异均有统计学意义(P<0.05)。与模型组比较,各治疗组可下调血清IL-4、ECP的水平(P<0.01);与中药组和西药组比较,中西医结合组疗效最佳,IL-4、ECP差异有统计学意义(P<0.05)。中药组和中西医结合组可上调INF-γ表达水平(P<0.01);各治疗组可下调树突细胞表面CD80、CD86的表达水平(P<0.05);与中药组和西药组比较,中西医结合组疗效最佳,其CD80、CD86的表达水平差异有统计学意义(P<0.05)。结论温阳补肾方可以抑制哮喘小鼠气道的炎症反应,调节Th1及Th2免疫失衡,与地塞米松有协同效应,其下调脾脏树突细胞的表达是温阳补肾方治疗哮喘的一个重要机制。

Objective To investigate the immunoregulatory mechanism of ＂Wenyang Bushen Decoction＂ on spleen dendrictic cells in mice with asthma by observing the expression of costimulatory molecules CD50 and CD86 on mice spleen DC surface. Methods Thirty BALB/c mice were randomly divided into six groups： normal control group, model group, TCM group, western medicine group and integrative medicine group. Except for normal group, the others were intervened by corresponding measures. The inflammatory response of lung tissue in every group was observed after hematoxylin-eosin （HE） staining; the contents of IL-4, IFN-γ and ECP in the serum were detected by ELISA; separated the dendritic cells of mouse spleen tissue in each group and cultured the dendritic cells, and observed the DC morphology after scanned it. We analyzed the expression of costimulatory molecules CDS0 and CD86 on mouse spleen DC surface in each group by FACS. Results There was airway epithelial damage, loss, eosinophil infiltration in the model group. Compared with the normal group, the levels of IL- 4 and ECP were increased, the INF-γ level was decreased and the positive expressions of CDSO and CD86 were increased, with significant difference between normal group and model group. Compared with the model group, the levels of IL-4 and ECP in the other treatment groups were decreased （ P 〈 0.01 ） ; compared with TCM group and western medicine group, the integrative group had the best effect, and significant differences was found in IL-4 and ECP （P 〈 0.05 ） ; the Chinese medicine group and integrative group can up-regulate the expression of INF-γ （ P 〈 0.01 ） ; all the treatment groups can down-regulate the expression of CD80 and CD86 on cell surface （ P 〈 0.05 ） ; compared the TCM group and western medicine group, the integrative group had the best effect, with significant differences in expression of CDS0 and CD86 （P 〈 0.05 ）. Conclusion ＂Wenyang Bushen Decoction＂ can inhibit the inflammatory response of the airways in asthmatic mice, regulate immune imbalance of TH1 and TH2 and has synergistic effect with dexamethasone. ＂Wenyang Bushen Decoction＂ can down-regulate the expression of spleen dendritic cells, which is an important mechanism of it.