目的 探讨树突状细胞相关C型凝集素-1 (Dectin-1)在热处理光滑假丝酵母菌刺激大鼠气道上皮细胞(RTECs)中的作用及对IL-10和TNF-α表达的影响.方法 将体外培养的RTECs随机分为3组:对照组(RTECs+无菌生理盐水)、真菌刺激组(RTECs+热处理光滑假丝酵母菌)、抑制剂干预组(RTECs+昆布多糖+热处理光滑假丝酵母菌),分别孵育0、2、4、6h后终止,MTT法检测细胞存活率,Western Blot法检测Dectin-1表达,ELISA法检测IL-10和TNF-α的表达.结果 热处理光滑假丝酵母菌破坏细胞结构,降低细胞存活率.在培养开始时(0 h),3组RTECs细胞存活率以及Dectin-1、IL-10、TNF-α表达水平比较,差异均无统计学意义(均P＞0.05).培养2、4、6h后,真菌刺激组、抑制剂干预组Dectin-1、IL-10、TNF-α表达水平均高于对照组,抑制剂干预组Dectin-1、IL-10、TNF-α表达水平均低于真菌刺激组(均P＜0.05).除抑制剂干预组0h与2 h IL-10表达量比较,差异无统计学意义之外,真菌刺激组和抑制剂干预组组内不同时间段Dectin-1、IL-10、TNF-α表达水平比较,差异均有统计学意义(均P＜0.05).结论 Dectin-1是RTECs识别热处理光滑假丝酵母菌的重要受体,并诱导其释放IL-10和TNF-α,介导炎症反应的发生.%Objective To investigate the effect of Dectin-1 on the release of inflammatory factors interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α) in rat tracheal epithelial cells (RTECs) stimulated with heat-treated Candida glabrata (C.glabrata).Methods RTECs cultivated in vitro were randomly divided into three groups,including control group(RTECs + sterile normal saline),fungal stimulation group(RTECs + heat-treated C.glabrata),and inhibitor intervention group(RTECs + laminarin + heat-treated C.glabrata),cells were harvested after incubation for 0,2,4,6 hours respectively,cell survival rate was determined by MTT method,expression of Dectin-1 was analyzed by Western Blot,expression of IL-10 and TNF-α were detected by enzyme-linked immunosorbent assay (ELISA).Results Heat-treated C.glabrata destroyed cell structure and reduced cell survival rate.At the beginning of the culture (0 h),cell survival rate,expressions of Dectin-1,IL-10 and TNF-α among three groups were all not significantly different(all P＞0.05).After incubation for 2,4,6 hours,expressions of Dectin-1,IL-10 and TNF-α in fungal stimulation group and inhibitor intervention group were both significantly higher than control group;expressions of Dectin-1,IL-10,and TNF-α in inhibitor intervention group was lower than fungal stimulation group(all P＜0.05).The expressions of IL-10 in inhibitor intervention group at 0 h and 2 h was not significantly different,expressions of Dectin-1,IL-10,and TNF-α in fungal stimulation group and inhibitor intervention group at different incubation periods were significantly different(all P＜0.05).Conclusion Dectin-1 is an important receptor for RTECs to recognize the heat-treated C.glabrata,it induces the release of IL-10 and TNF-α,and mediate the occurrence of inflammation.
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