交通相关PM_(2.5)对人外周血淋巴细胞凋亡的影响  

Effects of Traffic Related PM_(2.5) on Apoptosis of Human Peripheral Blood Lymphocytes

作　　者：田家瑜 郭丽丽 邱勇[1,3] 乔果果 张志红 TIAN Jia-yu;GUO Li-li;QIU Yong;QIAO Guo-guo;ZHANG Zhi-hong(Department of Environmental Health,School of Public Health,Shanxi Medical University,Taiyuan 030001,China;Science and Education Department of Taiyuan Center for Disease Control and Prevention,Taiyuan 030001,China;Longquan Ping an Community Health Service Center,Longquanyi District,Chengdu 610100,China;Teaching and Experimental Center,School of Public Health,Shanxi Medical University,Taiyuan 030001,China)

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,太原030001 [2]太原市疾病预防控制中心科教科,太原030001 [3]成都市龙泉驿区龙泉平安社区卫生服务中心,成都610100 [4]山西医科大学公共卫生学院教学实验中心,太原030001

出　　处：《科学技术与工程》2021年第12期4856-4861,共6页Science Technology and Engineering

基　　金：国家自然科学基金(81872589,30700655,81072261);山西省应用基础研究计划项目(2018-319);山西省青年科技研究基金(2008021053)。

摘　　要：为探讨交通相关PM_(2.5)对人外周血淋巴细胞凋亡的诱导作用及可能机制,为交通相关PM_(2.5)的免疫毒性提供实验依据,采用0、50、100、200μg/mL PM_(2.5)对人外周血淋巴细胞进行24 h和48 h染毒,FITC-AnnexinV/PI染色,流式细胞仪检测淋巴细胞早期凋亡和坏死;采用0、20、80、320μg/mL PM_(2.5)浓度分别染毒人外周血淋巴细胞24、48、72 h,Western blot法测定细胞内含半胱氨酸的天冬氨酸特异蛋白水解酶3(cysteinyl aspartate specific proteinase 3,Caspase-3)和cAMP反应元件结合蛋白(cAMP-response element binding protein,CREB)蛋白表达水平,ELISA法检测细胞内环磷酸腺苷(cyclic adenosine monophosphate,cAMP)含量。结果表明,50、100、200μg/mL PM_(2.5)染毒细胞24 h和48 h后,淋巴细胞早期凋亡和坏死均高于生理盐水组,差异有统计学意义(P<0.01),且具有一定的剂量-效应关系;20、80、320μg/mL PM_(2.5)染毒细胞24、48、72 h后,各剂量组Caspase-3蛋白表达水平均高于对照组,除80μg/mL PM_(2.5)染毒72 h外,差异均具有统计学意义(P<0.01)。320μg/mL PM_(2.5)染毒细胞24、48、72 h后,细胞内cAMP含量均比生理盐水组升高(P<0.05)。PM_(2.5)染毒72 h后,CREB表达水平随PM_(2.5)浓度的升高而降低(P<0.01)。可见,交通相关PM_(2.5)可诱导人外周血淋巴细胞产生凋亡,且细胞内cAMP和CREB参与了凋亡的调控。To explore the inducing effect of traffic related PM_(2.5)on apoptosis of human peripheral blood lymphocytes and its possible mechanism,so as to provide experimental basis of immunotoxicity of traffic related PM_(2.5),Human peripheral blood lymphocytes were exposed to 0,50,100,200μg/mL PM_(2.5)for 24 h and 48 h and FITC-AnnexinV/PI staining and flow cytometry was used to detect the early apoptosis and necrosis of lymphocytes.The expression of cysteinyl aspartate specific proteinase 3(Caspase-3)and cAMP-response element binding protein(CREB)protein in human peripheral blood lymphocytes and the intracellular cyclic adenosine monophosphate(cAMP)content were measured by Western blot and ELISA after exposure to PM 2.5 at 0,20,80 and 320μg/mL for 24,48 and 72 h.The results show that after cells are exposed to 50,100 and 200μg/mL PM_(2.5)for 24 h and 48 h,the early apoptosis and necrosis of lymphocytes are higher than those in the normal saline group,with statistically significant differences(P<0.01),and there is a dose-effect relationship.The protein expression level of Caspase-3 in each dose group is higher than that in the control group after 24 h,48 h and 72 h of 20,80,320μg/mL PM_(2.5)exposure,and except for 80μg/ml PM 2.5 exposure for 72 h,the differences are statistically significant(P<0.01).After 24 h,48 h and 72 h of 320μg/mL PM_(2.5)exposure,the intracellular cAMP content is increased compared with the normal saline group(P<0.05).And the expression level of CREB decrease with the increase of PM_(2.5)concentration after exposure for 72 h(P<0.01).It is concluded that traffic related PM_(2.5)can induce apoptosis of human peripheral blood lymphocytes,and intracellular cAMP and CREB are involved in the regulation of apoptosis.

关 键 词：交通相关PM_(2.5) 淋巴细胞 凋亡 CASPASE-3 环磷酸腺苷 环磷腺苷反应元件结合蛋白 

分 类 号：R122.7[医药卫生—环境卫生学]