中华皮肤科杂志 ›› 2012, Vol. 45 ›› Issue (11): 792-795.

• 论著 • 上一篇    下一篇

热休克蛋白70上调寻常性银屑病成纤维细胞白介素6表达

尧志建1,王梅2,王香兰3,王煜4,任建文2,刘平5,王琼玉2   

  1. 1. 广西医科大学第一附属医院皮肤性病科
    2. 西安交通大学第二附属医院皮肤科
    3. 西安交通大学第二医院皮肤科
    4. 西安交通大学第二附属医院
    5. 西安交通大学医学院第二附属医院皮肤科
  • 收稿日期:2011-12-22 修回日期:2012-07-14 出版日期:2012-11-15 发布日期:2012-10-31
  • 通讯作者: 王梅 E-mail:wangmei6972@163.com
  • 基金资助:

    13115重大科技专项基金（抗IGF-1R银屑病治疗新药的研发）

Heat shock protein 70 upregulates interleukin-6 expression by fibroblasts from psoriasis vulgaris lesions

  • Received:2011-12-22 Revised:2012-07-14 Online:2012-11-15 Published:2012-10-31

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摘要：

目的 探讨热休克蛋白70（HSP70）对寻常性银屑病皮损中成纤维细胞白介素6（IL-6）表达的影响。 方法 原代培养出寻常性银屑病皮损中的成纤维细胞（PFb），然后在PFb培养基中加入5 ～ 30 mg/L的HSP70以及50 μmol/L的核因子κB特异性抑制剂吡咯啉烷二甲基硫脲（PDTC），并孵育不同时间（0 ～ 48 h）。以正常生长的PFb为对照组。用酶联免疫吸附法（ELISA）检测PFb培养上清液中IL-6的含量，用半定量逆转录-聚合酶链反应（RT-PCR）检测PFb IL-6 mRNA表达。运用t检验和Dunnett-t检验进行统计分析。结果 HSP70呈时间依赖（0 ～ 48 h）和剂量依赖（5 ～ 30 mg/L）性增加PFb IL-6蛋白合成和mRNA表达。10 mg/L HSP70实验组IL-6蛋白［（75.2 ± 15.4） ng/L］和IL-6 mRNA表达水平（0.439 ± 0.093）均显著高于对照组［分别为（47.2 ± 10.6 ）ng/L和0.249 ± 0.069］，两组比较，差异均有统计学意义（P ＜ 0.05）。30 mg/L HSP70实验组培养PFb 12 h时IL-6蛋白开始明显增多；培养6 h时IL-6 mRNA开始明显增多，与对照组相比，差异均有统计学意义（P ＜ 0.05）。随着HSP70浓度的增加和PFb培育时间的延长，HSP70实验组与对照组IL-6蛋白和IL-6 mRNA的表达差异逐渐加大。３０ mg/L HSP70 + PDTC组IL-6蛋白（42.23 ± 9.41 ng/L）和IL-6 mRNA表达（0.144 ± 0.048）均显著低于对照组（分别为68.40 ± 14.43 ng/L和0.295 ± 0.081），两组比较，差异均有统计学意义（P ＜ 0.05）。结论 HSP70能增强核因子κB介导的PFb IL-6蛋白和IL-6 mRNA表达，可能是诱导PFb高水平IL-6表达的因素之一。

关键词: 白细胞介素6

Abstract:

Objective To evaluate the in vitro effect of heat shock protein 70 （HSP70） on interleukin-6 （IL-6） expression by cultured fibroblasts from psoriasis vulgaris lesions（PFbs）. Methods Fibroblasts were isolated from the lesions of patients with psoriasis vulgaris and subjected to a primary culture. After 3 to 5 passages of culture, the fibroblasts were collected and used in the next experiment. Some PFbs were cultured with different concentrations （5, 10, 20, 30 mg/L） of HSP70 for 48 hours, or with HSP70 of 30 mg/L for different durations （3, 6, 12, 24, 48, 72 hours）; some PFbs were incubated with HSP70 of 30 mg/L for 24 hours after pretreatment with pyrrolidine dithiocarbamate（PDTC, a specific inhibitor of nuclear factor-kappa B） for 30 minutes. PFbs receiving no treatment served as the control. Enzyme-linked immunosorbent assay （ELISA） and semi-quantitative reverse transcription PCR were performed to measure the IL-6 protein expression in culture supernatant and IL-6 mRNA expression by PFbs, respectively. Differences in the expression of IL-6 protein and mRNA between PFbs receiving different treatment were analyzed by using t test and Dunnett′s t test. Results HSP70 significantly increased both protein production and mRNA expression of IL-6 in a time（0－48 h）- and dose（5－30 mg/L）-dependent manner. The expression levels of supernatant IL-6 protein and IL-6 mRNA were significantly higher in the PFbs treated with HSP70 of 10 mg/L for 48 hours than untreated PFbs （（75.2 ± 15.4） ng/L vs. （47.2 ± 10.6） ng/L, 0.439 ± 0.093 vs. 0.249 ± 0.069, both P ＜ 0.05）. A significant increase was observed as early as 6 hours in the level of IL-6 mRNA after the treatment with HSP70 of 30 mg/L, and 12 hours in the level of supernatant IL-6 protein. Decreased supernatant IL-6 protein and IL-6 mRNA were noted for PFbs treated with PDTC and HSP70 of 30 mg/L compared with untreated PFbs （（42.23 ± 9.41） ng/L vs. （68.40 ± 14.43） ng/L, 0.144 ± 0.048 vs. 0.295 ± 0.081, both P < 0.05）. Conclusion HSP70 may increase the expression of IL-6 mRNA and protein by cultured PFbs via the nuclear factor-kappa B pathway.

Key words: Interleukin-6

中图分类号: 

  • R751