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
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骆雪萍

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R519.3

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广西自然科学基金资助项目(2013GXNSFAA019209);广西医疗卫生重点科研课题(重2012007)


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

    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(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、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0 h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。

    Abstract:

    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.

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赵莹,骆雪萍,张雪,等. 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[J]. 中国感染控制杂志,2017,16(1):10-15. DOI:10.3969/j. issn.1671-9638.2017.01.003.
ZHAO Ying, LUO Xueping, ZHANG Xue, et al. Role of Dectin-1 in the production of IL-10 and TNF-α by rat tracheal epithelial cells stimulated with heattreated Candida glabrata[J]. Chin J Infect Control, 2017,16(1):10-15. DOI:10.3969/j. issn.1671-9638.2017.01.003.

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  • 收稿日期:2016-07-12
  • 最后修改日期:2016-09-12
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  • 在线发布日期: 2017-01-22
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