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

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P0.05)。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P0.05)。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P0.05)。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。

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

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.