烟曲霉孢子对肥大细胞活化作用的研究

摘要

Objective To study the etfects of Aspergillus fumigatus spore on the activation of human mast cell line HMC-1.Methods Prepared heat-killed Aspergillusfumigatus spore antigen by heating method.HMC-1 cells were co-cultured with heat-killed Aspergil lus fumigatus spores whose concentration was 5 × 105.Microplate spectrophotometer was applied to the β-hexosaminidase release assay which was an index of mast cell degranulation.Enzyme-linked immunosorbent assay (ELISA) was used for released IL-5.The expression levels of IL-5 mRNA was detected by Realtime-PCR.Results Compared with the negative control group,heat-killed Aspergillus fumigatus spores increased β-hexosaminidase release [(11.3 ％ ± 1.2 ％) VS (0.5％±0.1％),P＜0.05],IL-5 mRNA expression level [(2.47±0.12) VS (0±0.26),P＜0.05] and the release of IL-5 [(11.96±1.09) VS (9±1.36),P＜0.05] in HMC-1 cells.Conclusion Aspergillusfumigatus spores activates mast cell line HMC-1 resulting in cell degranulation,synthesis and release of IL-5.%目的 研究烟曲霉孢子对人肥大细胞系HMC-1的活化作用.方法 用加热灭活的方法制备烟曲霉孢子抗原,体外培养人肥大细胞系HMC-1细胞,检测热灭活烟曲霉孢子诱导HMC-1细胞脱颗粒β-氨基己糖苷酶释放比,酶联免疫吸附测定(ELISA)法检测HMC-1细胞培养上清中白细胞介素-5(IL-5)的释放,Realtime-PCR检测热灭活烟曲霉孢子诱导的HMC-1细胞1L-5 mRNA的表达变化.结果 与阴性对照组相比,热灭活烟曲霉孢子刺激后HMC-1细胞的β-氨基己糖苷酶释放比升高[(11.3％±1.2％)VS(0.5％±0.1％),P＜0.05],IL-5释放增加[(11.96土1.09)VS(9.00± 1.36),P＜0.05],IL-5mRNA表达上调[(2.47±0.12)VS(0.00±0.26),P＜0.05].结论 热灭活烟曲霉孢子诱导HMC-1肥大细胞活化,引起细胞活化脱颗粒、合成和释放IL-5增加.