余艳琴, 郝金奇, 姚三巧, 张春民, 陈志远, 白玉萍, 徐应军, 沈福海, 车审言, 范雪云, 袁聚祥. TNF-α/TNFR信号蛋白在煤工尘肺患者肺泡巨噬细胞中的表达[J]. 环境与职业医学, 2016, 33(7): 665-669. DOI: 10.13213/j.cnki.jeom.2016.15474
引用本文: 余艳琴, 郝金奇, 姚三巧, 张春民, 陈志远, 白玉萍, 徐应军, 沈福海, 车审言, 范雪云, 袁聚祥. TNF-α/TNFR信号蛋白在煤工尘肺患者肺泡巨噬细胞中的表达[J]. 环境与职业医学, 2016, 33(7): 665-669. DOI: 10.13213/j.cnki.jeom.2016.15474
YU Yan-qin, HAO Jin-qi, YAO San-qiao, ZHANG Chun-min, CHEN Zhiyuan, BAI Yu-ping, XU Ying-jun, SHEN Fu-hai, CHE Shen-yan, FAN Xue-yun, YUAN Ju-xiang. Expression of TNF-α/TNFR-Associated Signaling Proteins in Alveolar Macrophages in Coal Workers with Pneumoconiosis[J]. Journal of Environmental and Occupational Medicine, 2016, 33(7): 665-669. DOI: 10.13213/j.cnki.jeom.2016.15474
Citation: YU Yan-qin, HAO Jin-qi, YAO San-qiao, ZHANG Chun-min, CHEN Zhiyuan, BAI Yu-ping, XU Ying-jun, SHEN Fu-hai, CHE Shen-yan, FAN Xue-yun, YUAN Ju-xiang. Expression of TNF-α/TNFR-Associated Signaling Proteins in Alveolar Macrophages in Coal Workers with Pneumoconiosis[J]. Journal of Environmental and Occupational Medicine, 2016, 33(7): 665-669. DOI: 10.13213/j.cnki.jeom.2016.15474

TNF-α/TNFR信号蛋白在煤工尘肺患者肺泡巨噬细胞中的表达

Expression of TNF-α/TNFR-Associated Signaling Proteins in Alveolar Macrophages in Coal Workers with Pneumoconiosis

  • 摘要: 目的

    研究肺泡巨噬细胞(alveolar macrophage, AM)凋亡信号通路中肿瘤坏死因子α(TNF-α)/肿瘤坏死因子受体(TNFR)信号蛋白的表达水平及其规律,探讨TNF-α/TNFR信号蛋白在煤工尘肺肺纤维化过程中的可能作用。

    方法

    以尘肺诊断中确定的观察对象(4例)及壹期煤工尘肺(CWP)患者(24例,其中脱尘及未脱尘各12例)为研究对象,收集大容量肺灌洗回收液,经分离、纯化、培养裂解肺泡巨噬细胞,AM用蛋白免疫印迹(Western blotting)检测信号蛋白β-actin、TNF-α、TNFR1、TNFR2、天冬半胱氨酸特异性蛋白酶8(Caspase-8)、天冬半胱氨酸特异性蛋白酶3(Caspase-3)、核因子-κB的受体65/50(NF-κB P65及P50),以β-actin为参比,用FlourChem HD2软件分析各信号蛋白的相对含量。

    结果

    壹期CWP患者AM表达的TNF-α、TNFR1、Caspase-8、Caspase-3、NF-κB(P65、P50)、TNFR2水平分别为2.69±1.19、1.95± 1.02、2.80±1.95、2.19±1.49、2.70±1.30、2.45±1.32、5.15±1.24,高于观察对象中其相应蛋白(分别为0.93±0.14、1.18± 0.19、1.24±0.55、1.44±1.08、1.41±0.26、1.35±0.53、1.07±0.08)的表达,差异具有统计学意义(P < 0.05);在未脱尘者中上述指标表达水平分别是3.544±1.278、2.508±0.925、3.274±2.183、3.079±1.304、3.332±1.062、2.865±1.024、5.817±1.089,高于脱尘者中的表达水平(分别为1.899±1.191、1.318±0.538、1.627±0.740、1.764±0.815、2.068±0.708、1.602±0.672、4.483±1.425),差异具有统计学意义(P < 0.05)。

    结论

    CWP患者AM凋亡信号蛋白Caspase-8、Caspase-3、NF-κB(P65、P50)、TNFR、TNF-α的表达水平上升,脱尘后表达水平降低。表明TNF-α/TNFR信号通路在尘肺发病中可能起一定作用。

     

    Abstract: Objective

    To study the expression level and pattern of tumor necrosis factor α(TNF-α)/tumor necrosis factor receptor(TNFR) signaling proteins in apoptosis signal pathway in alveolar macrophage(AM), and investigate the possible role of TNF-α/TNFR signaling proteins in promoting pulmonary fibrosis in coal workers with pneumoconiosis.

    Methods

    Coal workers under observation(n=4) and with Stage I coal workers' pneumoconiosis(CWP) n=24, including 12 cases diagnosed after being removed from dust exposed workstation(RDE) and 12 cases diagnosed during working in dust exposed workstation(DE) were enrolled as study subjects. Large volume pulmonary lavage fluids were collected, isolated, and purified for AM lysis. Western blot was used to detect β-actin, TNF-α, TNFR1, TNFR2, Caspase-8, Caspase-3, nuclear factor kappa B(NF-κB) P65 and P50, using β-actin as reference. Relative expression levels of each signaling protein was analyzed using FlourChem HD2 software.

    Results

    The expression levels of TNF-α, TNFR1, Caspase-8, Caspase-3, NF-κB P65, NF-κB P50, and TNFR2 in the CWP stage I patients were 2.69±1.19, 1.95±1.02, 2.80±1.95, 2.19±1.49, 2.70±1.30, 2.45±1.32, and 5.15±1.24, respectively, higher than those of the observation workers(0.93±0.14, 1.18±0.19, 1.24±0.55, 1.44±1.08, 1.41±0.26, 1.35±0.53, and 1.07±0.08)(P < 0.05). In the DE workers, the expression levels were 3.544±1.2783, 2.508±0.925, 3.274±2.183, 3.079±1.304, 3.332±1.062, 2.865±1.024, and 5.817±1.089, respectively, higher than those of the RDE workers(1.899±1.191, 1.318±0.538, 1.627±0.740, 1.764±0.815, 2.068±0.708, 1.602±0.672, and 4.483±1.425)(P < 0.05).

    Conclusion

    Elevated expression levels of AM apoptosis signaling proteins including Caspase-3, Caspase-8, NF-κB(P65 and P50), TNFR, and TNF-α are detected in CWP patients, and decreased after workers being removed from dust exposed workstation. The TNF-α/TNFR signaling pathway may play a part in pneumoconiosis occurrence.

     

/

返回文章
返回