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目的培养支气管哮喘患儿外周血树突状细胞,体外给予CRTH2受体拮抗剂干预,从而探讨CRTH2受体在支气管哮喘发病中的作用。方法提取外周血单个核细胞,体外给于10 ng/ml GM-CSF及IL-4的完全培养液培养,并在第5天磁珠分选得到纯度90%以上的树突状细胞,体外给予CRTH2受体拮抗剂干预,收集细胞及上清,运用流式细胞技术检测DCs表型,ELISA方法检测细胞上清IL-10、IL-12含量变化,同种混合淋巴细胞反应检测树突状细胞刺激淋巴细胞增殖及分化的能力。结果经磁珠分选后,树突状细胞纯度可达90%,符合实验要求;给药组与对照组相比,给药组外周血树突状细胞表面表达的CD80、CD86、CD40显著增高(P<0.05);给药组外周血树突状细胞分泌的IL-12明显高于对照组,IL-10明显低于对照组(P<0.05);给药组外周血树突状细胞刺激T淋巴细胞增殖能力显著大于对照组(P<0.05);给药组外周血树突状细胞刺激T淋巴细胞分泌IL-4含量明显低于对照组,IFN-γ含量明显高于对照组(P<0.05)。结论与对照组相比,CRTH2受体可能通过调节树突状细胞的功能来达到参与疾病的发生发展。
Objective To culture peripheral blood dendritic cells from children with bronchial asthma and to investigate the effect of CRTH2 receptor on the pathogenesis of bronchial asthma by intervention of CRTH2 receptor antagonist in vitro. Methods Peripheral blood mononuclear cells (PBMCs) were extracted and cultured in complete medium containing 10 ng / ml GM-CSF and IL-4. The dendritic cells with the purity over 90% CRTH2 receptor antagonist intervention, the cells and supernatants were collected, the DCs phenotype was detected by flow cytometry, the content of IL-10 and IL-12 in the supernatant of the cells was detected by ELISA, the dendritic cells The ability to stimulate lymphocyte proliferation and differentiation. Results The dendritic cells could reach 90% purity by magnetic beads sorting, which accorded with the experimental requirements. Compared with the control group, the expression of CD80, CD86 and CD40 on the surface of dendritic cells was significantly increased (P <0.05). IL-12 secreted by peripheral blood dendritic cells in the drug-treated group was significantly higher than that in the control group, and IL-10 was significantly lower than that in the control group (P <0.05). The dendritic cells stimulated (P <0.05). The content of IL-4 secreted by peripheral blood dendritic cells stimulated by T lymphocytes in the treated group was significantly lower than that in the control group, and the content of IFN-γ in the T lymphocyte was significantly higher than that in the control group <0.05). Conclusion Compared with the control group, CRTH2 receptor may be involved in the development of the disease by regulating the function of dendritic cells.