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[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Flour Chem 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。
[Objective] To investigate the expression of tumor necrosis factor α (TNF-α) / tumor necrosis factor receptor (TNFR) signaling protein in alveolar macrophage (AM) Possible role of TNFR signaling protein in pulmonary fibrosis in coal worker ’s pneumoconiosis. [Method] With the objects identified in the diagnosis of pneumoconiosis (4 cases) and the one phase of coal worker’s pneumoconiosis (CWP) (24 cases, of which 12 cases were de-dusting and not de-dusting) The recovered liquid was isolated, purified and cultured to lyse alveolar macrophages. AM was used to detect the expression of signal protein β-actin, TNF-α, TNFR1, TNFR2, aspartate cysteine specific protease 8 ( Caspase-8, Caspase-3, NF-κB P65 and P50, β-actin was used as a reference, Flour Chem HD2 software analyzes the relative amounts of each signal protein. [Results] The levels of TNF-α, TNFR1, Caspase-8, Caspase-3, NF-κB (P65, P50) and TNFR2 in the first phase of CWP were 2.69 ± 1.19, 1.95 ± 1.02, 2.80 ± 1.95 and 2.19 ± 1.49, 2.70 ± 1.30, 2.45 ± 1.32 and 5.15 ± 1.24, respectively, higher than the corresponding proteins in the observed subjects (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), the difference was statistically significant (P <0.05). The expression levels of the above indexes in the non-dedusting were 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, which were higher than those in the de-agglomerates (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 respectively) Statistically significant (P <0.05). [Conclusion] The expression levels of apoptosis signal protein Caspase-8, Caspase-3, NF-κB (P65, P50), TNFR and TNF-α in CWP patients were increased, It indicated that the TNF-α / TNFR signaling pathway may play a role in the pathogenesis of pneumoconiosis.