交通相关细颗粒物对人外周血淋巴细胞免疫毒性和钙信号机制研究  

Immunotoxicity of Traffic Related Fine Particles to Human Peripheral Blood Lymphocytes and Calcium Signal Mechanism

作　　者：郭丽丽[1] 张志红[1] 原福胜[1] 董洁[1] 乔果果[1] 

机构地区：[1]山西医科大学公共卫生学院环境卫生学教研室,山西太原030001

出　　处：《环境与健康杂志》2010年第11期946-949,共4页Journal of Environment and Health

基　　金：国家自然科学基金资助项目(30700655);山西省青年科技研究基金资助项目(2008021053);山西省高校科技研究开发项目(2007124);山西医科大学大学生创新基金资助项目(200629;200981)

摘　　要：目的探讨交通相关的细颗粒物(PM2.5)对人外周血淋巴细胞的免疫毒性,从钙信号途径揭示免疫抑制机制。方法采用0、5、20、80、320、800μg/ml PM2.5染毒T淋巴细胞24、48 h,采用四甲基偶氮唑盐(MTT)比色法测定T淋巴细胞的增殖功能。采用0、50、100、200μg/ml PM2.5染毒T淋巴细胞244、8 h,采用流式细胞仪测定T淋巴细胞的免疫功能(CD3+/CD4+/CD8+亚群和CD4+/CD8+)。采用荧光分光光度计测定T淋巴细胞的钙离子浓度([Ca2+]i)。用试剂盒测定T淋巴细胞的Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活力。结果 320、800μg/ml PM2.5可抑制细胞增殖功能,且与对照组比较有统计学意义(P<0.05);50、200μg/ml PM2.5染毒细胞24 h,染毒组T淋巴细胞亚群(CD3+、CD4+百分含量、CD4+/CD8+比值)与对照组比较变化不明显;50、100、200μg/ml PM2.5染毒细胞48 h后,染毒组T淋巴细胞亚群(CD3+、CD4+百分含量、CD4+/CD8+比值)均低于对照组,且各剂量组CD3+和200μg/ml CD4+百分含量与对照组比较,差异有统计学意义(P<0.05)。200μg/mlPM2.5染毒细胞1、3、61、2、24 h后[,Ca2+]i均高于对照组,除24 h外,与对照比较,差异均具有统计学意义(P<0.05)。在3 h时胞内[Ca2+]i最高,随时间延长[,Ca2+]i逐渐降低。100、200μg/ml PM2.5染毒细胞3 h时,[Ca2+]i高于对照组,差异有统计学意义(P<0.05);随染毒浓度的升高,Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活力有降低趋势,且Na+-K+-ATP酶活力在200μg/mlPM2.5组与对照组比较,差异有统计学意义(P<0.05)。结论交通相关的PM2.5可引起人外周血淋巴细胞免疫抑制,其可能与钙稳态失衡有关。Objective To know the immunotoxicity of fine particles to human peripheral blood lymphocytes and to explore the calcium signaling pathway related immunosuppression mechanism.Methods The cells were exposed to related PM2.5 at the doses of 0,5,20,80,320,800 μg/ml for 24 h and 48 h respectively,and the proliferation function was determined by MTT.At 0,50,100,200 μg/ml,exposed for 24 and 48 h,the lymphocyte CD3＋/CD4＋/CD8＋ subsets,CD4＋/CD8＋ were measured by flow cytometry.At 0,50,100,200 μg/ml,exposed for 24 h and 48 h,the i change in cells was detected by fluorescence spectrophotometry.The activities of Na＋-K＋-ATP enzyme and Ca2＋-Mg2＋-ATP enzyme were determined by the relative kit.Results The SI of lymphocyte decreased significantly in 320 and 800 μg/ml groups（P0.05）.T-lymphocyte subsets（CD3＋,CD4＋ percentage levels,CD4＋/CD8＋ ratio） in 50,100,200 μg/ml groups exposed for 48 h were significantly lower compared with the control group（P0.05）.After 1,3,6,12 and 24 h of PM2.5 exposure,except 24 h,the lymphocyte i levels in 200 μg/ml groups were higher than that in the control groups（P0.05）.The i levels after 3 h of PM2.5 exposure was the highest,the i levels gradually reduced with the exposure went on.The lymphocyte i level increased with the exposure concentration after 3h of exposure.As the PM2.5 exposure increased,the activities of Na＋-K＋-ATP enzyme and Ca2＋-Mg2＋-ATP enzyme decreased.Conclusion Traffic related PM2.5 can cause immunosuppression of human peripheral blood lymphocytes,the imbalance of calcium homeostasis may be concerned in the mechanism.

关 键 词：PM2.5 淋巴细胞 钙 三磷酸腺苷酶 

分 类 号：R994.6[医药卫生—毒理学]