论文部分内容阅读
目的探讨煤工尘肺(CWP)患者肺泡巨噬细胞(AM)凋亡的肿瘤坏死因子(TNF)-α及其受体(TNFR)信号转导通路调控作用机制。方法采用单纯随机抽样方法,以24例CWP壹期患者为CWP组,以4例煤矿粉尘作业观察对象人员为观察对象组,收集2组人员的支气管肺泡灌洗液分离、纯化AMs,分为对照组、超氧化物歧化酶(SOD)组、TNF/TNFR组、抗-TNF-α抗体组、半胱氨酸天冬氨酸蛋白酶(Caspase)-8抑制组和核转录因子(NF)-κB抑制组,进行相应处理后培养24 h,收获细胞,提取蛋白采用免疫印迹法检测信号蛋白TNF-α、TNFR1、TNFR2、Caspase-8、Caspase-3、NF-κB P50及NF-κB P65的表达情况。结果 CWP组AMs的TNF-α、TNFR2、Caspase-8、Caspase-3、NF-κB P50及NF-κB P65的蛋白相对表达水平均高于观察对象组(P<0.05)。TNF/TNFR组和抗-TNF-α抗体组AMs的TNF-α、TNFR1、Caspase-8、Caspase-3、NF-κB P50的蛋白相对表达水平均低于对照组(P<0.05);抗-TNF-α抗体组AMs的上述5个指标均低于NF-κB抑制组(P<0.05)。TNF/TNFR组AMs的TNFR1、Caspase-8和Caspase-3的蛋白相对表达水平均高于SOD组和Caspase-8抑制组(P<0.05),TNFR1、Caspase-8和NF-κB P50的蛋白相对表达水平均低于NF-κB抑制组(P<0.05)。对于CWP患者,TNF/TNFR组AMs的TNFR2和NF-κB P65的蛋白相对表达水平均低于对照组(P<0.05),高于SOD组(P<0.05)。结论 TNF/TNFR/NF-κB信号转导途径介导的AM凋亡在CWP的发病中起重要作用;对TNF/TNFR/NF-κB信号转导通路的不同环节进行抑制,可影响CWP患者凋亡蛋白的表达。
Objective To investigate the regulatory mechanism of tumor necrosis factor (TNF) -α and its receptor (TNFR) signal transduction pathway in alveolar macrophage (AM) apoptosis in coal worker’s pneumoconiosis (CWP) patients. Methods A simple random sampling method was used in this study. Twenty-four patients with CWP-1 were treated with CWP and 4 patients with dust-exposed coal mine workers were selected as the observation group. The bronchoalveolar lavage fluid from two groups was collected and purified for AMs. (SOD) group, TNF / TNFR group, anti-TNF-α antibody group, caspase-8 inhibition group and nuclear transcription factor (NF) -κB The cells were harvested and cultured for 24 h. The protein levels of TNF-α, TNFR1, TNFR2, Caspase-8, Caspase-3, NF-κB P50 and NF-κB P65 were detected by Western blotting Happening. Results The relative expression levels of TNF-α, TNFR2, Caspase-8, Caspase-3, NF-κB P50 and NF-κB P65 in CWs were higher than those in the observation group (P <0.05). The relative expression levels of TNF-α, TNFR1, Caspase-8, Caspase-3 and NF-κB P50 in AMs were lower than those in control group (P <0.05) The above five indexes of AMs in TNF-α antibody group were lower than those in NF-κB inhibition group (P <0.05). The relative expression levels of TNFR1, Caspase-8 and Caspase-3 in AMs in TNF / TNFR group were significantly higher than those in SOD and Caspase-8 inhibition groups (P <0.05), and the relative expressions of TNFR1, Caspase-8 and NF- The expression level of NF-κB was lower than that of NF-κB inhibition group (P <0.05). For CWP patients, TNFR2 and NF-κB P65 protein expression of TNF-α / TNFR group was lower than that of control group (P <0.05), but higher than that of SOD group (P <0.05). Conclusion AM apoptosis induced by TNF / TNFR / NF-κB signal transduction pathway plays an important role in the pathogenesis of CWP. Inhibition of different aspects of TNF / TNFR / NF-κB signaling pathway may affect CWP apoptosis Death protein expression.