内质网应激凋亡相关基因mRNA在矽肺患者肺泡巨噬细胞表达研究  

Expression of gene mRNA related to endoplasmic reticulum stress-mediated apoptosis in alveolar macrophages of patients with silicosis

作　　者：孙海霞[1] 姚三巧[1,2] 韩亚凤[1] 何艳玲[1] 陈刚[3] 朱丽[3] 金玉兰[1] 白玉萍[1] 郝小惠[4] 刘和亮[4] 

机构地区：[1]河北联合大学公共卫生学院,河北唐山063000 [2]新乡医学院公共卫生学院,河南新乡453000 [3]中国煤矿工人北戴河疗养院,秦皇岛066100 [4]河北联合大学医学实验研究中心,063000

出　　处：《中国职业医学》2015年第3期274-279,共6页China Occupational Medicine

基　　金：河北省自然科学基金(H2014209114)

摘　　要：目的探讨矽肺患者肺泡巨噬细胞(AM)中内质网应激(ERS)凋亡相关基因的表达规律和意义。方法采用单纯随机抽样方法,选择矽肺壹期、贰期和叁期男性患者23、25、24例分别设为矽肺壹期、贰期、叁期组,以8例诊断为观察对象的男性矽尘作业工人为对照组,收集研究对象的支气管肺泡灌洗液,分离、纯化和培养AM后,提取RNA,采用实时定量荧光聚合酶链式反应分析AM中葡萄糖调节蛋白78(GRP78)、CAAT区/增强子结合蛋白同源蛋白(CHOP)、天冬氨酸特异性半胱氨酸蛋白酶(Caspase)-4和Caspase-3的mRNA表达水平。结果矽肺壹期、贰期和叁期组患者AM中GRP78、CHOP、Caspase-4和Caspase-3的mRNA相对表达水平均高于对照组(P<0.05),矽肺贰期组GRP78 mRNA相对表达水平分别高于矽肺壹期和叁期组(P<0.05),矽肺贰期和叁期组患者AM中的CHOP、Caspase-4和Caspase-3的mRNA相对表达水平均高于矽肺壹期组(P<0.05)。多重逐步线性回归分析结果显示,矽肺患者AM中GRP78、CHOP、Caspase-4和Caspase-3的mRNA相对表达水平均与矽肺期别呈正相关(P<0.05)。结论 ERS在矽肺患者AM凋亡的发生与发展中可能发挥重要作用。Objective To investigate the expression regulation and significance of apoptosis-related gene mRNA with endoplasmic reticulum stress（ ERS） in alveolar macrophages（ AM） of patients with silicosis. Methods Using simple random sampling method,the male patients with silicosis at stages Ⅰ,Ⅱ,and Ⅲ were separately chosen as the silicosis groupⅠ,Ⅱ,and Ⅲ with 23,25,and 24 cases in each group. Eight male workers diagnosed as observed subjects exposed to silica dust were selected as the control group. Bronchoalveolar lavage fluids were collected from all subjects. After AMs were separated,purified,and cultured,the mRNA expression levels of glucose regulated protein 78（ GRP78）,CAAT region / enhancer binding protein homologous protein（ CHOP）,Caspase-4,and Caspase-3 were measured by Real Time PCR after total RNA was extracted. Results The mRNA relative expression levels of GRP78,CHOP,Caspase-4,and Caspase-3 in AMs of silicosis groups Ⅰ,Ⅱ,and Ⅲ were higher than those of the control group respectively（ P〈0. 05）,the relative expression level of GRP78 mRNA in AMs of silicosis groupⅡ was higher than those of silicosis groups Ⅰ andⅢ（ P〈0. 05）. Relative expression levels of CHOP,Caspase-4,and Caspase-3 mRNA in AMs of silicosis groups Ⅱ andⅢ were higher than those of the silicosis group Ⅰ（ P〈0. 05）. Multiple stepwise linear regression indicated that GRP78,CHOP,Caspase-4,and Caspase-3 mRNA expression levels in AMs of silicosis patients were positively correlated to the stages of silicosis（ P〈0. 05）. Conclusion ERS may play an important role in the occurrence and development of AM apoptosis in patients with silicosis.

关 键 词：矽肺 细胞凋亡 肺泡巨噬细胞 内质网应激 葡萄糖调节蛋白78 CAAT区/增强子结合蛋白同源蛋白 天冬氨酸特异性半胱氨酸蛋白酶 

分 类 号：R135.2[医药卫生—劳动卫生]