蛋白激酶C／核因子κB信号通路在燃煤砷致人体T细胞活化中的作用  

Role of protein kinase C/NF-kappa B signal pathway in activation of T lymphocytes in human exposed to coal-arsenic

作　　者：谷俊莹[1] 张爱华[1] 张碧霞[2] 黄晓欣[2] 

机构地区：[1]贵阳医学院毒理学教研室,550004 [2]中国人民解放军第44医院

出　　处：《中国地方病学杂志》2010年第1期33-36,共4页Chinese Jouranl of Endemiology

基　　金：基金项目：教育部博士点基金（G2007-05）;贵州省科技厅课题（黔科合重大专项字[200616016号）;贵州省“十一五”攻关项目（黔科合NY字[2006]3057号）志谢衷心感谢解放军第44医院董学新、杨大平、岑笃才以及贵阳医学院公共卫生学院梁冰、张然、于春、王清青等在课题实施过程中给予的帮助

摘　　要：目的了解燃煤砷暴露人群外周血T细胞活化情况及其外周血单个核细胞（PBMCs）内蛋白激酶C／核因子κB（PKC／NF—κB）途径相关信号分子的表达或活化情况，探讨PKC／NF—κB信号通路在燃煤砷致人体T细胞活化中的作用。方法按照《地方性砷中毒诊断标准》（WS／T211-2001）将贵州省兴仁县交乐村燃煤污染型地方性砷中毒病区94例居民分为无症状暴露组（12例）、轻度砷中毒组（33例）、中度砷中毒组（34例）及重度砷中毒组（15例）；以距病区约12km的非砷污染某村居民27例作为对照组。采用流式细胞术检测外周血活化T细胞比例：电泳迁移率改变实验（EMSA）检测PBMCs内NF—κB结合DNA的活性；蛋白印迹法检测PBMCs胞浆内PKC0及磷酸化蛋白激酶CO（pPKC0）表达水平。结果外周血活化T细胞比例，各组间比较差异有统计学意义（F=7．893，P〈0．01），无症状暴露组[（21．76±15．31）％]及轻、中、重度砷中毒组[（18．41±11．36）％、（17．78±11．93）％和（18．79±13．38）％]与对照组[（3．19±2．12）％]比较明显增加（P〈0．05或〈0．01）：无症状暴露组及轻、中、重度砷中毒组PBMCs内NF—κB结合DNA的活性（1．49±0．24、1．58±0．30、1．57±0．34、1．51±0．16）与对照组（1．304-0．17）比较，差异有统计学意义（P〈0．05或〈0．01）。PBMCs胞浆PKC0表达水平，组间比较差异无统计学意义（F=1．843，P〉0．05）。pPKC0表达水平，组间比较差异有统计学意义（F=6．588，P〈0．01），轻、中、重度砷中毒组（0．64±0．14、0．64±0．20、0．62±0．12）与对照组（O．93±0．20）及无症状暴露组（O．81±0．23）比较，明显降低（P〈0．05或〈0．01）。PBMCs内pPKC0表达水平与NF-KB结合DNA的活性呈明显负相关（r=-0．565，P〈0．01）。NF—κB结合DNA的活性与外周血活化T细胞比例呈明显正相关（r=0�Objective To investigate the activation of T lymphocytes in human peripheral blood and the signaling molecules in protein kinase C/nuclear factor κB （PKC/NF-κB） pathway expressivity or activity changes in human peripheral blood mononuclear cells （PBMCs） exposed to coal-arsenic, to explore the role of PKC/NF-κB signal pathway in activation of T cells in human exposed to coal-arsenic. Methods Blood samples were collected from individuals who lived in arsenism area of coal-burning in Guizhou province, and were divided into asymptomatically exposed group （12）, mild arsenism group （33）, moderate arsenism group （34） and severe arsenism group（15） according to Diagnosis Standard for Endemic Arsenism（WS/T 211-2001 ）. The individuals who lived in non-arsenism area were control group（27）. The ratio of activated T cells was analyzed by flow cytometry. DNA binding activity of NF-KB in PBMCs was evaluated by electrophoretic mobility shift assay（EMSA）. The expression of PKC0 and phospho-PKC0 （pPKC0） in PBMCs were detected with western blotting analysis. Results The ratio of activatingT cells in asymptomatically exposed group [ （21.76 ± 15.31 ） % 1, mild arsenism group [ （18.41 ± 11.36） % ], moderate arsenism group [ （ 17.78 ± 11.93）% ] and severe arsenism group [ （18.79 ± 13.38） % 1 were all higher than that of control group[（3.19 ± 2.12）%], the difference among all groups being statistically significant （F = 7.893, P 〈 0.05）. DNA binding activity of PBMCs NF-κB in asymptomatically exposed group, mild arsenism group, moderate arsenism group and severe arsenism group（ 1.49 ± 0.24,1.58 ± 0.30,1.57 ± 0.34,1.51 ± 0.16） were higher than that of the control group（l.30 ± 0.17）, the difference being statistically significant（P 〈 0.05 or 〈 0.01）. The expression of PBMCs pPKC0 in mild arsenism group, moderate arsenism group and severe arsenism group（0.64 ± 0.14,0.64 ± 0.27, 0.62 ± 0.12） were all lower than that of the control group（0.

关 键 词：砷 煤 T淋巴细胞 蛋白激酶C 核因子ΚB 

分 类 号：R686[医药卫生—骨科学]