Fonsecaea monophora对巨噬细胞TLR2、TLR4、Dectin-1和TNF-α表达的影响

中国真菌学杂志　 2014, Vol. 9 　Issue (3): 134-138.

Fonsecaea monophora对巨噬细胞TLR2、TLR4、Dectin-1和TNF-α表达的影响

蒋丽¹, 张军民¹, 孙九峰², 席丽艳¹, 蔡文莹¹, 肖星¹

1. 中山大学孙逸仙纪念医院皮肤科, 广州 510120;
2. 广东省疾病预防控制中心, 广州 510120

收稿日期:2014-03-28 出版日期:2014-06-28 发布日期:2014-06-28
通讯作者: 张军民，E-mail：junminmx@163.com E-mail:junminmx@163.com
作者简介:蒋丽，女（汉族），硕士研究生在读.E-mail：327492551@qq.com
基金资助:
广东省自然科学基金（S2013010015189），国家自然科学基金（81301411）

Eeffect of Fonsecaea monophora on the expressions of TLR2,TLR4,Dectin-1 and the production of TNF-α in macrophages

JIANG Li¹, ZHANG Jun-Min¹, SUN Jiu-Feng², XI Li-Yan¹, CAI Wen-Ying¹, XIAO Xing¹

1. Department of Dermatology, Sun Yat-Sen Hospital, Sun Yat-Sen University, Guangzhou 510120, China;
2. Centre for Disease Prevention and Control of Guangdong Province, Guangzhou 510120, China

Received:2014-03-28 Online:2014-06-28 Published:2014-06-28

摘要/Abstract

摘要：

目的探讨F.monophora对巨噬细胞模式识别受体TLR2、TLR4及Dectin-1的表达及介导炎症因子TNF-α分泌的影响。方法 F.monophora分生孢子突变株（色素株，CBS122845）及白化突变株（CBS125149）菌液分别与小鼠巨噬细胞（RAW264.7）共培养，RAW264.7巨噬细胞作为阴性对照。采用Real Time PCR检测各组TLR2、TLR4、Dectin-1和下游信号分子MyD88、NF-κB mRNA的表达情况，流式细胞术检测TLR2、TLR4、Dectin-1蛋白的表达水平，ELISA法检测培养上清中TNF-α因子的分泌情况。结果与对照组比较，CBS122845和CBS125149组中巨噬细胞TLR2、TLR4、Dectin-1和MyD88、NF-κB的mRNA和蛋白表达量均升高；与CBS125149比较，CBS122845组中TLR2的mRNA和蛋白的表达量无明显变化，而TLR4、Dectin-1和MyD88、NF-κB的mRNA和蛋白的表达量升高更明显；与对照组比较，CBS122845组培养上清TNF-α因子的分泌量明显减低，CBS125149组TNF-α因子的分泌量明显升高。结论 TLR2、TLR4、Dectin-1这三个模式识别受体可能共同参与巨噬细胞对F.monophora的识别过程，并可激活下游信号分子MyD88和NF-κB；且CBS122845可以通过抑制TNF-α分泌来抑制炎症反应，维持菌体在巨噬细胞中的长期生存和繁殖。

关键词: Fonsecaea monophora, Toll样受体2, Toll样受体4, 树突状细胞相关C型凝集素-1, 肿瘤坏死因子-&alpha

Abstract:

Objective To explore the effect of F.monophora on the expressions of toll-like receptor 2 (TLR2),toll-like receptor 4 (TLR4),C-type lectin-1 (Dectin-1) and the production of proinflammatory cytokine tumor necrosis factor-α (TNF-α) in macrophages.Method Murine macrophages (RAW264.7) were co-cultured with the meristematic mutant (CBS122845)and the albino mutant (CBS125149)of F.monophora. RAW264.7 macrophages were used as negative control. We detected the mRNA expressions of TLR2, TLR4, Dectin-1 and MyD88, NF-κB by real-time PCR (RT-PCR) and detected the protein expressions of TLR2, TLR4, Dectin-1 by Flow CytoMetry (FCM). The protein expression of TNF-α was measured by ELISA.Results Compared with the negative control group, the mRNA and protein expressions of TLR2, TLR4, Dectin-1 and MyD88, NF-κB increased in CBS122845 and CBS125149 group. Compared to CBS125149 group, the mRNA and protein expression of TLR2 in CBS122845 group had no significant change, while the mRNA and protein expressions of TLR4, Dectin-1 and MyD88, NF-κB increased more significantly.Compared with the control group, CBS122845 group significantly decreased the protein secretion of TNF-α cytokine in culture supernatant, and CBS125149 group up-regulated the protein secretion of TNF-α cytokine obviously.Conclusion The pattern recognition receptors of TLR2, TLR4, Dectin-1 may participate in the recognization of F.monophora by macrophages,and activate the downstream signal molecules MyD88 and NF-κB;CBS122845 can inhibit the expression of TNF-α to maintain long-term growth and reproduction in macrophage.

Key words: Fonsecaea monophora, Toll-like receptor 2, Toll-like receptor 4, Dectin-1, Tumor necrosis factor-α

中图分类号:

R379.9

蒋丽, 张军民, 孙九峰, 席丽艳, 蔡文莹, 肖星. Fonsecaea monophora对巨噬细胞TLR2、TLR4、Dectin-1和TNF-α表达的影响[J]. 中国真菌学杂志, 2014, 9(3): 134-138.

JIANG Li, ZHANG Jun-Min, SUN Jiu-Feng, XI Li-Yan, CAI Wen-Ying, XIAO Xing. Eeffect of Fonsecaea monophora on the expressions of TLR2,TLR4,Dectin-1 and the production of TNF-α in macrophages[J]. Chinese Journal of Mycology, 2014, 9(3): 134-138.

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