百草枯对巨噬细胞的毒性作用及ROS、IL-6和TNF-α产生的影响

Toxicity Analysis of Paraquat on Macrophages and the Effects on Production of ROS,IL-6 and TNF-α

目的研究百草枯对巨噬细胞的毒性作用及对活性氧(ROS)、白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)产生的影响。方法不同浓度百草枯作用小鼠巨噬细胞RAW264.7,分别在1,2,8 h收取细胞和细胞培养液,检测细胞生存率及细胞内ROS(化学荧光法),细胞培养液IL-6、TNF-α(ELISA法)。结果在实验观察的8 h内,0.001、0.01、0.1 mmol/L百草枯作用的RAW264.7存活率没有明显变化,10 mmol/L和1 mmol/L百草枯作用其存活率分别在2 h和8 h开始下降。1 h时,随着百草枯浓度增加,细胞内ROS荧光强度递增;从2 h开始,1 mmol/L和10 mmol/L百草枯细胞内ROS荧光强度下降。随着百草枯浓度从0.001 mmol/L增加到1 mmol/L,细胞培养液中的IL-6的浓度递增;当百草枯浓度增加到10 mmol/L时,细胞培养液中的IL-6浓度下降。细胞培养液中的TNF-α浓度随着百草枯浓度增加和作用时间的延长而增加(除外10 mmol/L百草枯作用8 h)。结论高浓度的百草枯对巨噬细胞产生毒性作用,适当浓度的百草枯使巨噬细胞ROS、IL-6及TNF-α的产生增加。

Objective To study the effects of paraquat （PQ） on macrophages, including the toxicity and the induced production of reactive oxygen species （ ROS）, interleukin - 6 （ IL- 6 ） and tumor necrosis factor - α （TNF - α ）. Methods The RAW26d.7 macrophages were treated with different concentrations of PQ for 1 h, 2 h and 8 h. The cells and medium were harvested, and the cell viability, intracellular ROS （chemiluminescence method）, IL-6 and TNF-α in medium （ ELISA ） were measured. Results There was no significant difference of the cell viability among 0.001 mmol/L, 0.01 mmol/L and 0.1 mmol/L PQ treatment for 8 h, while the decreased cell survival was observed with 1 mmol/L and 10 mmol/L PQ treatment for 2 h and 8 h. The intracellular ROS fluorescent intensity increased at 1 h with a concentration dependent mode of PQ from 0.001 mmol/L to 10 mmol/L. ROS levels decreased with 1 mmol/L and 10 mmol/L PQ treatment for 2 h and 8 h. The IL-6 concentration in the supernatant increased with a con- centration dependent mode of PQ from 0.001 mmol/L to 1 mmol/L and decreased when PQ concentration increasing to 10 mmol/L. The TNF-α level in the supernatant increased with the PQ concentration increasing and the treating time extending except for 10 mmol/L PQ at 8 h. Conclusion High concentration of PQ shows toxic effect on macrophages, and appropriate concentrations of PQ increases the production of ROS, TNF-α and IL-6 by macrophages.