TNF-α/TNFR/NF-κB通路对煤工尘肺患者肺泡巨噬细胞凋亡调控机制  

The TNF-α/TNFR/NF-κB pathway in regulating alveolar macrophage apoptosis in coal workers' pneumoconiosis

作　　者：余艳琴[1,2] 郝金奇 姚三巧[3,2] 张春民 陈志远[5] 白玉萍[2,6] 陈刚[5] 张志浩[5] 徐应军[2] 袁扬[5] 沈福海 马国宣[5] 黄京慧 车审言[5] 范雪云[2] 袁聚祥[2] 

机构地区：[1]内蒙古科技大学包头医学院公共卫生学院,内蒙古包头014060 [2]华北理工大学公共卫生学院,河北唐山063000 [3]新乡医学院公共卫生学院,河南新乡453000 [4]国家安全生产监督管理总局职业安全卫生研究中心,北京102308 [5]中国煤矿工人北戴河疗养院尘肺科,秦皇岛066000 [6]河北省煤矿卫生与安全实验室,063000

出　　处：《中国职业医学》2017年第3期303-308,共6页China Occupational Medicine

基　　金：国家自然科学基金(30671741)

摘　　要：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 study the mechanism of tumor necrosis factor （TNF）-α and its receptor （TNFR） signal transduction pathways in regulating cell apoptosis of alveolar macrophage （AM） in coal workers＇＇ pneumoconiosis （CWP）.Methods Twenty-four coal workers with pneumoconiosis at stageⅠwere selected as CWP group and four observation subjects exposed to coal were chosen as observation group by using simple random sampling method.The bronchoalveolar lavage fluids of whole-lung lavage of two groups were collected.AMs were separated and purified.Then they were divided into 6 groups： a control group, a superoxide dismutase （SOD） group, a TNF/TNFR group, an anti-TNF-α antibody group, a Caspase-8 suppression group and a nuclear factor-κB （NF-κB） suppression group.The AMs of 6 groups with corresponding treatment were cultivated.After 24 hours, the cells were harvested and proteins extracted.The relative expression of TNF-α, TNFR1, TNFR2, Caspase-8, Caspase-3, NF-κB P50 and NF-κB P65 protein was detected by Western blotting.Results The protein relative expression of TNF-α, TNFR2, Caspase-8, Caspase-3, NF-κB P50 and NF-κB P65 in CWP group was significantly higher than those in the observation group （P〈0.05）.The protein relative expression of TNF-α, TNFR1, Caspase-8, Caspase-3 and NF-κB P50 in the TNF/TNFR group and the anti-TNF-α antibody group was lower than that of the control group （P〈0.05）.The above indexes in the anti-TNF-α antibody group were lower than that of the NF-κB suppression group （P〈0.05）.The protein relative expression of TNFR1, Caspase-8 and Caspase-3 in the TNF/TNFR group was higher than that of the SOD group and the Caspase-8 suppression group （P〈0.05）.The protein relative expression of TNFR1, Caspase-8 and NF-κB P50 in the TNF/TNFR group was lower than that of the NF-κB suppression group （P〈0.05）.Among the CWP patients, the relative expression of TNFR2 and NF-κB P65 in the TNF/TNFR group was lower than that of the control group （P�

关 键 词：煤工尘肺 肺泡巨噬细胞 信号通路 凋亡蛋白 肿瘤坏死因子-α 半胱氨酸天冬氨酸蛋白酶 核转录因子-κB 

分 类 号：R135.2[医药卫生—劳动卫生]