交通相关PM2.5对人外周血淋巴细胞凋亡的影响
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R122

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


Effects of traffic related PM2.5 on apoptosis of human peripheral blood lymphocytes
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The National Natural Science Foundation of China(81872589,30700655,81072261);Applied basic research project of Shanxi Province(2018-319);Shanxi Youth Science and Technology Research Fund(2008021053)

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    摘要:

    为探讨交通相关PM2.5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PM2.5的免疫毒性提供实验依据。采用0、50、100、200 g/ml PM2.5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、320 g/ml PM2.5浓度分别染毒人外周血淋巴细胞24 h、48 h和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、200 μg/ml PM2.5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、320 μg/ml PM2.5染毒细胞24 h、48 h和72 h后,各剂量组Caspase-3蛋白表达水平均高于对照组,除80 μg/ml PM2.5染毒72 h外,差异均具有统计学意义(P < 0.01)。320 μg/ml PM2.5染毒细胞24 h、48 h和72 h后,细胞内cAMP含量均比生理盐水组升高(P < 0.05)。PM2.5染毒72 h后,CREB表达水平随PM2.5浓度的升高而降低(P < 0.01)。可见,交通相关PM2.5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。

    Abstract:

    To explore the inducing effect of traffic related PM2.5 on apoptosis of human peripheral blood lymphocytes and its possible mechanism, so as to provide experimental basis of immunotoxicity of traffic related PM2.5. Human peripheral blood lymphocytes were exposed to 0, 50, 100, 200 g/ml PM2.5 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 Caspase-3 and CREB protein in human peripheral blood lymphocytes and the intracellular cAMP content was measured by Western blot and ELISA after exposure to PM2.5 at 0, 20, 80 and 320 g/ml for 24 h , 48 h and 72 h. The results show that after cells were exposed to 50, 100 and 200 g/ml PM2.5 for 24 h and 48 h, the early apoptosis and necrosis of lymphocytes were higher than those in the normal saline group, with statistically significant differences (P<0.01), and there was a dose-effect relationship. The protein expression level of Caspase-3 in each dose group was higher than that in the control group after 24 h, 48 h and 72 h of 20, 80, 320 μg/ml PM2.5 exposure, and except for 80g/ml PM2.5 exposure for 72 h, the differences were statistically significant (P<0.01). After 24 h, 48 h and 72 h of 320 g/ml PM2.5 exposure, the intracellular cAMP content was increased compared with the normal saline group (P<0.05). And the expression level of CREB decreased with the increase of PM2.5 concentration after exposure for 72 h (P<0.01). It is concluded that traffic related PM2.5 can induce apoptosis of human peripheral blood lymphocytes, and intracellular cAMP and CREB are involved in the regulation of apoptosis.

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田家瑜,郭丽丽,邱勇,等. 交通相关PM2.5对人外周血淋巴细胞凋亡的影响[J]. 科学技术与工程, 2021, 21(12): 4856-4861.
Tian Jiayu, Guo Lili, Qiu Yong, et al. Effects of traffic related PM2.5 on apoptosis of human peripheral blood lymphocytes[J]. Science Technology and Engineering,2021,21(12):4856-4861.

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  • 收稿日期:2020-10-18
  • 最后修改日期:2020-12-21
  • 录用日期:2020-12-21
  • 在线发布日期: 2021-05-17
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