RAW264.7小鼠巨噬细胞与阿萨希毛孢子菌相互作用后TNF-α、IL-1β和CCL3的表达变化  

Expression of TNF-α,IL-1β and CCL3 in RAW264.7 macrophages challenged with Trichosporon asahii

作　　者：丛林[1] 夏志宽[1] 李海涛[1] 杨蓉娅[1] 

机构地区：[1]北京军区总医院全军皮肤损伤修复研究所,北京100700

出　　处：《中国真菌学杂志》2015年第4期203-206,共4页Chinese Journal of Mycology

基　　金：国家自然科学基金(81271764;81472892)

摘　　要：目的研究RAW264.7小鼠巨噬细胞与T.asahii相互作用后TNF-α,IL-1β和CCL3的表达水平变化。方法RAW264.7细胞与T.asahii菌液共孵育3 h、6 h、9 h、12 h,对照组不加菌液。在各孵育时间点取上清液,采用双抗体夹心酶联免疫吸附试验(ELISA)试剂盒检测上清液中TNF-α、IL-1β和CCL3的含量。台盼蓝拒染法计各孵育时间点RAW264.7细胞的活细胞数。结果 RAW264.7细胞与T.asahii共孵育3 h、6 h、9 h、12 h后,TNF-α、IL-1β、CCL3的表达水平均明显高于对照组,有显著性差异(P<0.05)。3种细胞因子的表达水平随共孵育时间延长而增高,共孵育9 h的表达水平最高。RAW264.7细胞的活细胞数随共孵育时间延长而逐渐下降,共孵育12 h后的活细胞数量与其他各时间点有统计学差异(P<0.05)。结论 RAW264.7小鼠巨噬细胞面临T.asahii感染时可分泌TNF-α、IL-1β、CCL3等细胞因子,这些炎性细胞因子可能参与了抗T.asahii感染的免疫反应。Objective To explore the protein levels of TNF-α, IL-1β and CCL3 in RAW264.7 macrophages challenged with Trichosporon asahii （T.asahii）.Methods RAW264.7 macrophages were incubated with T.asahii or medium alone （control group） for 3, 6, 9, or 12 h, then the cytokine concentrations （TNF-α, IL-1β and CCL3） in the supernatants were determined using commercial ELISA kits.The numbers of viable RAW264.7 macrophages after incubated with T.asahii for 3, 6, 9, or 12 h were counted by using trypan blue staining method.Results The protein levels of TNF-α, IL-1β and CCL3 in the supernatants when RAW264.7 macrophages incubated with T.asahii for 3, 6, 9, or 12 h were significantly higher than those in control group （P〈0.05）.The protein levels of these 3 cytokines were gradually elevated with the extension of incubation time and peaking at 9 h.The numbers of viable RAW264.7 macrophages were gradually declined with the extension of incubation time.The numbers of viable RAW264.7 macrophages were significantly declined after incubated with T.asahii for 12 h （P〈0.05）.Conclusion RAW264.7 macrophages could secrete TNF-α, IL-1β and CCL3 when challenged with T.asahii and these inflammatory cytokines may be involved in the anti-fungal immune response of RAW264.7 macrophages against T.asahii infection.

关 键 词：巨噬细胞 阿萨希毛孢子菌 细胞因子 

分 类 号：R379.9[医药卫生—病原生物学]