中华皮肤科杂志 ›› 2014, Vol. 47 ›› Issue (11): 803-805.

• 论著 • 上一篇    下一篇

白念珠菌对人THP-1细胞产生白细胞介素6、活化信号分子IκBα的影响

杨海平1,段志敏2,杜蕾蕾3,曾荣3,沈永年2,胡素泉4,刘维达2,陈青5,李岷2   

  1. 1. 南京解放军454医院皮肤科
    2. 南京 中国医学科学院北京协和医学院皮肤病研究所
    3. 中国医学科学院皮肤病研究所
    4. 中国医学科学院北京协和医学院皮肤病研究所
    5. 江苏省血液中心
  • 收稿日期:2014-01-26 修回日期:2014-09-12 出版日期:2014-11-15 发布日期:2014-11-01
  • 通讯作者: 李岷 E-mail:drlimin@sina.cn
  • 基金资助:
    白念珠菌细胞壁磷脂甘露聚糖与识别受体TLR2 在诱导固有免疫耐受中的作用及机制研究

Effect of Candida albicans on the production of interleukin-6 by and activation of IκBα in human THP-1 monocytes

  • Received:2014-01-26 Revised:2014-09-12 Online:2014-11-15 Published:2014-11-01

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摘要： 目的 探讨白念珠菌对人急性单核细胞白血病细胞系（THP-1细胞系）分泌白细胞介素6（IL-6）和细胞内信号分子NF-κB抑制蛋白（IκBα）激活的影响。 方法 实时荧光定量PCR分析105、106 CFU/ml灭活白念珠菌刺激THP-1细胞IL-6 mRNA表达水平变化，酶联免疫吸附法检测IL-6分泌量。免疫印迹法分析白念珠菌体外作用THP-1细胞后不同时间IκBα和磷酸化IκBα的水平。 结果 105 CFU/ml白念珠菌刺激THP-1细胞后1、3、6 h，IL-6 mRNA水平（2-ΔΔCt）分别为1.48 ± 0.06、6.48 ± 0.30、125.34 ± 1.47，刺激3 h、6 h后明显高于空白对照组（P < 0.001）。106 CFU/ml白念珠菌刺激THP-1细胞后1、3、6 h，IL-6 mRNA水平分别为2.96 ± 0.35、8.57 ± 1.27、588.10 ± 2.31，与空白对照组比较，P值分别为0.036、0.001、 < 0.001。106 CFU/ml白念珠菌刺激THP-1细胞后24 h，IL-6蛋白水平为（924.9 ± 30.13） ng/L，与空白对照组比较差异有统计学意义（P < 0.001）。106 CFU/ml白念珠菌作用THP-1细胞后30 min、60 min, 磷酸化IκBα蛋白水平显著升高，而IκBα蛋白水平相应降低。 结论 人THP-1细胞体外与白念珠菌作用后激活信号分子NF-κB并分泌IL-6，参与抗念珠菌感染固有免疫反应。

关键词: 念珠菌，白色, 白血病，单核细胞，急性, 白细胞介素6, I-κB蛋白质类, NF-κB

Abstract: Yang Haiping*, Du Leilei, Zeng Rong, Duan Zhimin, Shen Yongnian, Hu Suquan, Liu Weida, Chen Qing, Li Min. *454th Hospital of PLA, Nanjing 210002, China Corresponding authors: Li Min, Email: drlimin@sina.cn; Chen Qing, Email: qngchen@hotmail.com 【Abstract】 Objective To investigate the effect of Candida albicans on the production of interleukin-6 by and activation of IκBα in an acute monocytic leukemia cell line THP-1. Methods THP-1 cells were classified into three groups to be stimulated by heat-killed C. albicans cells in concentrations of 105 and 106 colony-forming units （CFU）/ml and lipopolysaccharide （100 μg/L） in vitro respectively. Those remaining untreated served as the blank control group. After additional culture for different durations, real time reverse transcription PCR and enzyme-linked immunosorbent assay were performed to measure the mRNA and protein expression levels of interleukin-6 （IL-6） respectively, and Western blot was conducted to determine the levels of total and phosphorylated IκBα. Statistical analysis was carried out by t test. Results A significant increase was observed in the mRNA expression level （2-ΔΔCt） of IL-6 in THP-1 cells at 3 and 6 hours after starting treatment with C. albicans at 105 CFU/ml （6.48 ± 0.30 vs. 0.84 ± 0.16, 125.34 ± 1.47 vs. 1.22 ± 0.22, both P < 0.01）, and at 1, 3 and 6 hours after starting treatment with C. albicans at 106 CFU/ml （2.96 ± 0.35 vs. 1.03 ± 0.16, 8.57 ± 1.27 vs. 0.84 ± 0.16, 588.10 ± 2.31 vs. 1.22 ± 0.22, P < 0.05 or 0.01） compared with the blank control group, but no significant difference was noted between THP-1 cells at 1 hour after starting treatment with C. albicans at 105 CFU/ml and those remaining untreated （1.48 ± 0.06 vs. 1.03 ± 0.16, P > 0.05）. The protein expression level of IL-6 was （924.9 ± 30.13） ng/L in THP-1 cells at 24 hours after starting treatment with C. albicans at 106 CFU /ml, significantly different from that in the blank control group （P < 0.001）. There was a marked elevation in the level of phosphorylated IκBα protein, but an obvious reduction in the level of IκBα protein in THP-1 cells treated with C. albicans at 106 CFU/ml for 30 and 60 minutes. Conclusion Human THP-1 monocytes may play a role in innate immune responses against C. albicans through activation of nuclear factor-κB and production of IL-6.

Key words: Candida albicans, Leukemia, monocytic, acute, Interleukin-6, I-kappa B proteins, NF-kappa B