周倩, 张兆强, 林立. 大鼠矽肺纤维化进程中肺巨噬细胞IL-1βIL-17CTGF mRNA表达的研究[J]. 环境与职业医学, 2020, 37(2): 174-180. DOI: 10.13213/j.cnki.jeom.2020.19525
引用本文: 周倩, 张兆强, 林立. 大鼠矽肺纤维化进程中肺巨噬细胞IL-1βIL-17CTGF mRNA表达的研究[J]. 环境与职业医学, 2020, 37(2): 174-180. DOI: 10.13213/j.cnki.jeom.2020.19525
ZHOU Qian, ZHANG Zhao-qiang, LIN Li. Expressions of IL-1β, IL-17, and CTGF mRNA in pulmonary macrophages in rats with silicosis fibrosis[J]. Journal of Environmental and Occupational Medicine, 2020, 37(2): 174-180. DOI: 10.13213/j.cnki.jeom.2020.19525
Citation: ZHOU Qian, ZHANG Zhao-qiang, LIN Li. Expressions of IL-1β, IL-17, and CTGF mRNA in pulmonary macrophages in rats with silicosis fibrosis[J]. Journal of Environmental and Occupational Medicine, 2020, 37(2): 174-180. DOI: 10.13213/j.cnki.jeom.2020.19525

大鼠矽肺纤维化进程中肺巨噬细胞IL-1βIL-17CTGF mRNA表达的研究

Expressions of IL-1β, IL-17, and CTGF mRNA in pulmonary macrophages in rats with silicosis fibrosis

  • 摘要: 背景

    矽肺是由于长期吸入游离二氧化硅(SiO2)含量较高的生产性粉尘而引起的以肺组织弥漫性纤维化为主的全身性疾病。该病是目前我国发病率及患病率最高的职业性疾患,但其发病机制尚未阐明。近年来,有关巨噬细胞合成和分泌的细胞因子在矽肺纤维化进程中的作用受到关注。探讨肺巨噬细胞相关细胞因子的作用,可能是揭示矽肺纤维化机制的重要方面。

    目的

    研究矽肺纤维化进程中大鼠肺巨噬细胞白细胞介素1β(IL-1β)、白细胞介素17(IL-17)、结缔组织生长因子(CTGF)mRNA的表达及其改变。

    方法

    将60只Wistar大鼠随机分为1个对照组和5个染尘组(15、30、60、90、120 d染尘组),每组10只。各染尘组大鼠经乙醚麻醉后,一次性肺内注射1mL矽尘悬浊液(100mg·mL-1),分别进行为期15、30、60、90、120 d的染尘实验。对照组大鼠经乙醚麻醉后灌注等体积生理盐水。染尘实验结束后,通过肺泡灌洗的方式获取肺巨噬细胞。取肺组织进行HE和Masson染色,同时测定各组肺组织羟脯氨酸(HYP)含量和肺巨噬细胞IL-1βIL-17CTGF mRNA的表达量。

    结果

    染色结果显示:15 d染尘组大鼠肺泡结构略有破坏、胶原纤维略有增多;随着染尘时间的延长,肺组织纤维化逐渐明显;120d染尘组大鼠整个肺组织呈弥漫纤维化改变。各组大鼠肺组织HYP含量随着染尘时间延长而增加,与对照组相比差异有统计学意义(P < 0.05),且呈线性上升趋势(P < 0.05)。对照组和15、30、60、90、120 d染尘组大鼠肺巨噬细胞IL-1βmRNA相对表达量分别为1.00±0.44、1.22±0.65、2.58±0.54、3.93±0.78、7.11±1.14、16.89±0.85;IL-17 mRNA相对表达量分别为1.00±0.36、4.25±2.49、7.00±2.95、14.75±3.95、37.28±2.51、50.27±5.66;CTGF mRNA相对表达量分别为1.00±0.48、3.87±0.29、4.97±1.20、11.08±0.60、15.50±2.71、23.64±4.11。除15 d染尘组大鼠肺巨噬细胞IL-1β mRNA表达量外,各组各细胞因子的mRNA表达量与对照组相比,差异均有统计学意义(P < 0.05),且呈线性上升趋势(P < 0.05)。

    结论

    矽尘可使肺巨噬细胞IL-1βIL-17CTGF mRNA的表达上调,且随染尘时间延长而增加,可能是矽肺纤维化的机制之一。

     

    Abstract: Background

    Silicosis is a systemic disease featured by diffuse fibrosis of lung tissues caused by chronic inhalation of occupational dust containing high-level free SiO2. This is also an occupational disease with the highest incidence and prevalence rates in China. However, the pathogenesis of silicosis is still unclear. In recent years, the cytokines related to macrophage synthesis and secretion have gained much attention owing to their significant role in the development of silicosis fibrosis. Exploring the role of cytokines related to pulmonary macrophages may be an important part to reveal the mechanism of silicosis fibrosis.

    Objective

    This experiment is conducted to investigate the expressions and changes of interleukin-1β (IL-1β), interleukin-17 (IL-17), and connective tissue growth factor (CTGF) mRNA in pulmonary macrophages in rats developing silicosis fibrosis.

    Methods

    A total of 60 Wistar rats were randomly divided into six groups with 10 rats in each group:one control group and five exposure groups (15, 30, 60, 90, and 120 d exposure groups). The rats in the exposure groups were injected of 1 mL silica dust suspension (100 mg·mL-1) for one time by pulmonary injection after anesthesia, and the observations lasted for 15, 30, 60, 90, and 120 d, respectively. The rats in the control group were perfused with normal saline of the same volume after anesthesia. Pulmonary macrophages were obtained by alveolar lavage after designed observation time ended. The lung tissues of each group were subject to HE and Masson staining, and determined for hydroxyproline (HYP) levels in liver tissues and expressions of IL-1β, IL-17, and CTGF mRNA in pulmonary macrophages.

    Results

    According to the staining results, the alveolar structure was slightly damaged and the collagen fibers were slightly increased in the 15 d exposure group; the pulmonary fibrosis was gradually obvious with the extension of silica dust exposure time; the whole lung showed diffuse fibrosis in the 120 d exposure group. Compared with the control group, the HYP levels in lung tissues were increased with the prolongation of exposure time (P < 0.05), and showed a rising linear trend (P < 0.05). The relative expression levels of IL-1β mRNA in the control group and five (15, 30, 60, 90, and 120 d) exposure groups were 1.00±0.44, 1.22±0.65, 2.58±0.54, 3.93±0.78, 7.11±1.14, and 16.89±0.85, respectively; the relative expression levels of IL-17 mRNA were 1.00±0.36, 4.25±2.49, 7.00±2.95, 14.75±3.95, 37.28±2.51, and 50.27±5.66, respectively; the relative expression levels of CTGF mRNA were 1.00±0.48, 3.87±0.29, 4.97±1.20, 11.08±0.60, 15.50±2.71, and 23.64±4.11, respectively. Compared with the control group, the mRNA expressions of selected cytokines in other exposure groups were significantly different (P < 0.05) and showed a linear rising trend (P < 0.05), except the IL-1β mRNA expression in the 15 d exposure group.

    Conclusion

    Silica dust could induce increased expressions of IL-1β, IL-17, and CTGF mRNA in pulmonary macrophages with the prolongation of exposure time, which might be one of the mechanisms of silicosis fibrosis.

     

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