摘要
将PM_(2.5)按照轻(L-PM_(2.5))、重(H-PM_(2.5))污染季节分为2类,分别按低、中、高3个剂量浓度染毒HBE细胞24 h后,检测其活性氧(ROS)、丙二醛(MDA)、谷胱甘肽(GSH)和超氧化物歧化酶(SOD)水平,探讨不同PM_(2.5)致HBE细胞氧化损伤及其机制。在评估重组人金属硫蛋白Ⅲ(rh-MT-Ⅲ)对HBE细胞毒性的基础上,将rh-MT-Ⅲ和N-乙酰半胱氨酸(NAC)分别预处理HBE细胞2 h后,高剂量下用2种PM_(2.5)继续暴露24 h,检测经抗氧化剂处理后的ROS、MDA、GSH含量和SOD活性,探究2种抗氧化剂对PM_(2.5)诱导的氧化损伤的干预作用。氧化损伤实验结果显示,经L-PM_(2.5)暴露后在中、高剂量下发现细胞中MDA含量的上升以及GSH含量和SOD活性的下降,H-PM_(2.5)可显著升高HBE细胞的ROS、MDA的水平,降低细胞中GSH含量和SOD活性,2种PM_(2.5)均能诱导HBE细胞发生氧化应激。细胞毒性结果显示,随着rh-MT-Ⅲ浓度的增加,HBE细胞存活率呈现下降的趋势,与对照组相比,在400μg·mL^(-1)实验浓度下HBE细胞存活率显著降低(P<0.05)。干预研究结果显示,rh-MT-Ⅲ可以显著降低H-PM_(2.5)暴露后细胞的ROS和MDA水平,升高细胞GSH含量,SOD活性无明显改变,但在L-PM_(2.5)染毒下,rh-MT-Ⅲ仅可改善MDA水平。NAC预处理后,细胞氧化损伤得到部分缓解,如MDA水平下降,GSH含量升高,但SOD活性没有明显变化。2种抗氧化剂在降低ROS和MDA水平及升高GSH含量方面存在差异,rh-MT-Ⅲ可以显著降低PM_(2.5)诱导的细胞ROS和MDA水平,NAC可显著提高GSH含量。PM_(2.5)暴露后可诱导HBE细胞氧化损伤,2种抗氧化剂可在一定程度上缓解这种氧化损伤。
HBE cells were exposed to low,medium and high concentrations of PM_(2.5)for 24 h,and the levels of r eactive oxygen species(ROS),malondialdehyde(MDA),glutathione(GSH)and superoxide dismutase(SOD)of HBE cells were detected to explore the oxidative damage and toxicological mechanisms.Based on the evaluation of the toxicity of the recombinant human metallothionein Ⅲ(rh-MT-Ⅲ)on HBE cells,HBE cells were pretreated with rh-MT-Ⅲ and N-acetylcysteine(NAC)for 2 h,then the cells were further exposed to the two types of PM_(2.5)for 24 h at high doses.The levels of ROS,MDA,GSH,and SOD were detected to explore the intervention effects of the two antioxidants on oxidative damage induced by PM_(2.5).The results of oxidative damage experiment showed that after exposure to L-PM_(2.5),MDA levels in HBE cells increased and GSH concentrations and SOD activity decreased.H-PM_(2.5)can significantly increase the levels of ROS and MDA in HBE cells,and decrease the concentrations of GSH and the activity of SOD.Both types of PM_(2.5)can induce oxidative stress in HBE cells.The cytotoxicity results showed that with the increase of rh-MT-Ⅲ dose,the viability of HBE cells showed a decreasing trend,and compared with the control group,the viability of HBE cells was significantly reduced at the dose of 400μg·mL^(-1)(P<0.05).The results of the intervention study showed that rh-MT-Ⅲ could significantly reduce the levels of ROS and MDA of HBE cells after exposure to H-PM_(2.5),and increase the concentration of GSH,and the activity of SOD had no significant change.However,rh-MT-Ⅲonly improved MDA levels of HBE cells when exposed to L-PM_(2.5).After NAC pretreatment,oxidative damage in HBE cells was partially alleviated,such as a decrease in MDA levels and an increase in GSH concentration,but there was no significant change in SOD activity.There were significant differences between the two antioxidants in reducing ROS and MDA levels,and increasing GSH concentration.rh-MT-Ⅲ could significantly reduce the levels of ROS and MDA of HBE cells,while NAC can significantly increase GSH concentration.Exposure to PM_(2.5)can induce oxidative damage in HBE cells,and the two antioxidants could alleviate the oxidative damage.
作者
甘俊英
沈鑫
伍勇
叶梦娟
邱光文
周燕
李艳艳
薛玉英
GAN Junying;SHEN Xin;WU Yong;YE Mengjuan;QIU Guangwen;ZHOU Yan;LI Yanyan;XUE Yuying(Maanshan Center for Disease Control and Prevention,Maanshan 243000,China;School of Public Health,Southeast University,Nanjing 210009,China)
出处
《生态毒理学报》
北大核心
2025年第3期524-533,共10页
Asian Journal of Ecotoxicology
基金
马鞍山市科技计划项目(YL-2021-26)。
