【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。更多还原

【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 Flour Chem 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.更多还原