Dectin-1在热处理光滑假丝酵母菌刺激大鼠气道上皮细胞中的作用及对IL-10和TNF-α表达的影响

Role of Dectin-1 in the production of IL-10 and TNF-α by rat tracheal epithelial cells stimulated with heat-treated Candida glabrata

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、6 h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(0h),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、6 h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。

ObjectiveTo 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）.MethodsRTECs 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）.ResultsHeat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）.ConclusionDectin-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.