申克孢子丝菌,单核细胞白血病,肿瘤坏死因子α,NF-κB ," /> 申克孢子丝菌,单核细胞白血病,肿瘤坏死因子α,NF-κB ,"/> sporothrix schenckii,monocytic leukemia,TNF-α,NF-κB ,"/> <p class="MsoNormal"> <span> <p class="MsoNormal"> <span>申克孢子丝菌酵母相对人急性单核细胞白血病细胞</span>NF-<span>κ</span><span>B</span><span>信号通路及</span><span>TNF-</span><span>α的影响</span> </span><span></span>

中国麻风皮肤病杂志 ›› 2017, Vol. 33 ›› Issue (2): 74-78.

• 论著 • 上一篇    下一篇

申克孢子丝菌酵母相对人急性单核细胞白血病细胞NF-κB信号通路及TNF-α的影响

刘彩霞1  段志敏1  杜蕾蕾1  曾荣1  沈永年1  胡素泉1  刘维达1  陈青2  李岷1   

  1. 1中国医学科学院北京协和医学院皮肤病研究所，江苏省皮肤性病分子生物学重点实验室,南京，210042

    2江苏省血液中心,南京，210042

  • 出版日期:2017-02-15 发布日期:2018-12-13
  • 通讯作者: 李岷，E-mail:drlimin@sina.cn 陈青，E-mail: qngchen@hotmail.com

Effects of 〖WTHX〗Sporothrix schenckii〖WTHZ〗 yeasts on the activation of NF-κB signal pathway and secretion of TNF-α in human acute monocytic leukemia cells

LIU Caixia1, DUAN Zhimin1, DU Leilei1, ZENG Rong1, SHEN Yongnian1, HU Suquan1, LIU Weida1, CHEN Qing2, LI Min1.   

  1. 1. Institute of Dermatology, Chinese Academy of Medical Sicences and Peking Union Medical College, Jiangsu Key Laboratory of Molecular Biology for Skin Diseases, Nanjing 210042, China; 2. Jiangsu Province Blood Center, Nanjing 210042, China

  • Online:2017-02-15 Published:2018-12-13
  • Contact: LI Min, E-mail: drlimin@sina.cn CHEN Qing, E-mail: qngchen@hotmail.com

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摘要： 目的：明确申克孢子丝菌酵母相对人急性单核细胞白血病细胞(THP-1)NF-κB信号通路激活和肿瘤坏死因子α(TNF-α)分泌的影响。方法：实时荧光定量PCR分析申克孢子丝菌酵母相刺激THP-1细胞TNF-α mRNA的表达，酶联免疫吸附法检测TNF-α分泌量，免疫印迹法分析申克孢子丝菌酵母相体外作用于THP-1细胞后IκBα和磷酸化IκBα的水平，免疫荧光法观察NF-κB-p65核转位。同时设置地塞米松（NF-κB抑制剂）抑制组，并检测100 nM地塞米松预处理THP-1细胞后TNF-α mRNA水平的变化。结果：申克孢子丝菌酵母相刺激THP-1细胞后6 h TNF-α mRNA水平显著高于空白对照组（P<0.001）。申克孢子丝菌酵母相刺激THP-1细胞后24 h，TNF-α蛋白水平为(4610.419±121.501)pg/mL，显著高于空白对照组(186.964 ± 98.073)pg/mL，差异具有统计学意义(P<0.001)。申克孢子丝菌酵母相作用THP-1细胞后30~60 min，磷酸化IκBα蛋白水平显著升高，为时间依赖性。申克孢子丝菌酵母相组细胞核内NF-κB-p65较空白对照组荧光强度增强。100 nM地塞米松预处理各组THP-1细胞后，TNF-α mRNA水平较前明显降低。结论：人THP-1细胞体外与申克孢子丝菌酵母相作用后激活NF-κB信号通路并上调TNF-α分泌。

关键词: 申克孢子丝菌')">

申克孢子丝菌, 单核细胞白血病, 肿瘤坏死因子α, NF-κB

Abstract:

Objective: To determine the effects of Sporothrix schenckii yeasts on the activation of NF -κB signal pathway and secretion of TNF-α in human acute monocytic leukemia cells (THP-1). Methods: The expression of TNF-α mRNA was detected by Real-time fluorescence quantitative PCR and enzyme-linked immunosorbent assay respectively. The level of phosphorylated IκBα was detected by Western blot. NF-κB-p65 nuclear translocation was measured by immunofluorescence. The level of TNF-α mRNA in THP-1 pretreated with 100 nM Dexamethasone (a NF-κB inhibitor) for 30 minutes was detected by Real-time fluorescence quantitative PCR. Results: The levels of TNF-α mRNA in THP-1 cells treated with Sporothrix schenckii yeasts for 6 hours were increased compared with the blank control group (P<0.001). The secretion level of TNF-α in the Sporothrix schenckii yeasts group was 4610.419±121.501 pg/mL, which was higher than that in the blank group (186.964±98.073 pg/mL), with a significant difference (P<0.001). Phosphorylation IκBα protein increased obviously and in a time -dependent manner after treated with Sporothrix schenckii yeasts from 30 minutes to 60 minutes. The fluorescent intensity of NF-κB-p65 in the Sporothrix schenckii yeasts group was stronger than that in the blank group. The level of TNF-α mRNA was decreased in the THP-1 macrophages treated with 100 nM dexamethasone in the three groups. Conclusion: Sporothrix schenckii yeasts can increase the expression of TNF-α through enhancing the activation of NF-κB pathway.

Key words: sporothrix schenckii')">

sporothrix schenckii, monocytic leukemia, TNF-α, NF-κB