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大气PM2.5颗粒物对大鼠心肌细胞的毒性作用研究
焦周光, 胡凌飞, 李娜, 张柯, 温占波, 李劲松
军事科学院军事医学研究院微生物流行病研究所
摘要:
为探讨大气PM2.5及其不同组分对心肌细胞H9C2的毒性作用,并评估PM2.5颗粒物对心血管系统产生毒性作用的关键组分。将前期采集并制备的PM2.5完全颗粒物、PM2.5水溶性组分、PM2.5脂溶性组分和PM2.5单纯颗粒物以不同浓度对H9C2细胞染毒。用MTS法在染毒6、10、24、48、72h后测定细胞活力;根据细胞活力测定结果,选用较低染毒浓度(10μg/mL),用相关试剂盒测定染毒24h后胞内和上清中LDH和SOD活力,ELISA及RT-QPCR法测定炎性因子IL-6和TNF-α表达量,AP位点计数法测定细胞DNA损伤情况。结果表明:颗粒物成分(PM2.5完全颗粒物和PM2.5单纯颗粒物)对H9C2细胞表现强烈生长抑制作用,而可溶性成分(PM2.5水溶性组分和PM2.5脂溶性组分)对H9C2细胞生长表现极弱或无生长抑制作用;各组分样本都在一定程度上破坏了H9C2细胞的胞膜通透性和抗氧化能力,PM2.5完全颗粒物和PM2.5脂溶性组分在造成炎性损伤方面作用明显。研究显示,颗粒物组分对H9C2细胞致死作用显著,PM2.5完全颗粒物相对来说表现出的毒性作用最强最全面。
关键词:  PM2.5  心肌细胞  细胞活力  炎性因子
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基金项目:国家自然科学基金项目(面上项目,重点项目,重大项目)
The Toxicity Effects Study on Rat Cardiac Myocytes from Atmospheric PM2.5 Particles
JIAO Zhouguang, HU Lingfei, LI Na, ZHANG Ke, WEN Zhanbo, LI Jinsong
State Key Laboratory of Pathogen and Biosecurity,Institute of Microbiology and Epidemiology,Academy of Military Medical Sciences,Academy of Military Science
Abstract:
In order to investigate the toxicity effects of atmospheric PM2.5 and its different fractions to the cardiac myocyte H9C2, and evaluate the key compositions of the toxicity effects of PM2.5 particles on the cardiovascular system. The prepared samples, such as original particles of PM2.5 (designated OPP2.5), water-soluble particles of PM2.5 (designated WPP2.5), fat-soluble particles of PM2.5 (designated FPP2.5) and pure particulate fractions of PM2.5 (designated PPP2.5), were used to exposed to H9C2 cells at different PM concentrations. The MTS method was used to test the cell viability at 6h, 10h, 24h, 48h, 72h post-treatment primarily, and according to the detected results of cell viability, a low PM concentration (10μg/mL) was selected for the follow-up experiment. And then the appropriate kits were used to detect the activities of LDH and SOD, ELISA and RT-QPCR were employed to detect the production of inflammatory cytokines IL-6 and TNF-α and the AP-sites counting was conducted to determine the level of intracellular DNA damage at 24h post-treatment. The results showed that, the insoluble samples (OPP2.5 and PPP2.5) showed strong inhibitory effects to cell growth, while the soluble samples (WPP2.5 and FPP2.5) showed very slight and even no inhibitory effects to cell growth; all samples destroyed the membrane permeability and antioxidant capacity of H9C2 cells to some extent, and OPP2.5 and FPP2.5 played an important role in induced inflammatory injury in H9C2 cells. The results showed that the insoluble samples of atmospheric PM2.5 showed significant lethal effect on H9C2 cells, and OPP2.5 showed strongest toxicity effects to H9C2 cells in all samples.
Key words:  PM2.5  Cardiac myocytes  Cell viability  Inflammatory factors