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目的探讨肿瘤坏死因子相关凋亡诱导配体(TRAIL)及其受体(TRAILR)信号转导通路在煤工尘肺(CWP)患者肺泡巨噬细胞(AM)凋亡中的作用机制。方法以接触煤尘及煤矽尘的24例观察对象和CWP壹期、贰期患者各22例为研究对象,收集大容量肺灌洗回收液,分离、纯化AM后,分为对照组、超氧化物歧化酶(SOD)组、TRAIL组、抗-TRAIL抗体组、天冬氨酸特异性半胱氨酸蛋白酶(Caspase)-8抑制组、核转录因子(NF)-κB抑制剂组,给予相应处理后进行培养。收获细胞后,检测细胞凋亡情况,并提取蛋白采用免疫印迹法检测TRAIL/TRAILR信号转导通路Caspase-3、Caspase-8和死亡受体5(DR5)蛋白相对表达水平。结果 5个处理组AM凋亡指数均低于对照组(P<0.05),TRAIL组AM凋亡指数分别高于SOD组、抗-TRAIL抗体组、Caspase-8抑制剂组和NF-κB抑制剂组(P<0.05);CWP壹期和贰期患者AM凋亡指数均高于观察对象(P<0.05),CWP贰期患者AM凋亡指数高于CWP壹期患者(P<0.05);不同处理和不同程度患者AM凋亡指数的交互作用有统计学意义(P<0.01)。5个处理组Caspase-3、Caspase-8和DR5的蛋白相对表达水平分别低于对照组(P<0.05),TRAIL组Caspase-3和Caspase-8的蛋白相对表达水平分别高于SOD组、抗-TRAIL抗体组、Caspase-8抑制剂组和NF-κB抑制剂组(P<0.05),SOD组DR5蛋白相对表达水平分别低于TRAIL组、抗-TRAIL抗体组、Caspase-8抑制剂组和NF-κB抑制剂组(P<0.05);CWP壹期和贰期患者Caspase-3、Caspase-8和DR5的蛋白相对表达水平均高于观察对象(P<0.05);不同处理和不同程度患者Caspase-3的蛋白相对表达水平的交互作用有统计学意义(P<0.01),不同处理和不同程度患者Caspase-8和DR5的蛋白相对表达水平的交互作用均无统计学意义(P>0.05)。结论 TRAIL/TRAILR信号转导通路介导的AM凋亡可能在CWP发病中发挥一定作用;在不同环节对TRAIL/TRAILR信号转导通路进行抑制或阻断,可有效抑制CWP患者AM凋亡的发生。
Objective To investigate the role of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its receptor (TRAILR) signal transduction pathway in alveolar macrophage (AM) apoptosis in coal worker’s pneumoconiosis (CWP) patients. Methods Twenty-four patients with CWS and 22 patients with stage I and II of CWP were enrolled in this study. A large volume of lung lavage recovery fluid was collected. After isolation and purification, AM was divided into control group, (SOD) group, TRAIL group, anti-TRAIL antibody group, caspase-8 inhibition group, nuclear factor-kappa B inhibitor group, The corresponding treatment after training. After the cells were harvested, the cell apoptosis was detected, and the proteins were extracted for the relative expression of TRAIL / TRAILR signal transduction pathway Caspase-3, Caspase-8 and death receptor 5 (DR5) protein by Western blotting. Results The apoptosis index of AM was lower in the five treatment groups than that in the control group (P <0.05). The apoptosis index of AM in TRAIL group was higher than that in SOD group, anti-TRAIL antibody group, Caspase-8 inhibitor group and NF- (P <0.05). The apoptosis index of AM in stage I and II of CWP was higher than that in observation stage (P <0.05), and the index of AM in stage II of CWP was higher than that in stage I of CWP (P <0.05) There was significant difference in the AM apoptosis index between the two groups (P <0.01). The relative expression levels of Caspase-3, Caspase-8 and DR5 in 5 treatment groups were lower than those in control group (P <0.05). The relative expression levels of Caspase-3 and Caspase-8 protein in TRAIL group were higher than those in SOD group TRAIL antibody group, Caspase-8 inhibitor group and NF-κB inhibitor group (P <0.05). The relative expression of DR5 protein in SOD group was lower than that in TRAIL group, anti-TRAIL antibody group and Caspase-8 inhibitor group (P <0.05). The relative expression levels of Caspase-3, Caspase-8 and DR5 in CWP stage I and II patients were significantly higher than those in the observation group (P <0.05) There was significant statistical significance in the relative expression of Caspase-3 protein (P <0.01). There was no significant difference in the relative expression levels of Caspase-8 and DR5 between different treatment groups and different degrees (P> 0.05) . Conclusions AM apoptosis induced by TRAIL / TRAILR signal transduction pathway may play a role in the pathogenesis of CWP. Inhibition or blocking of TRAIL / TRAILR signal transduction pathway at different stages can effectively inhibit the occurrence of AM apoptosis in CWP patients .