Integrated Systems Toxicology Division, National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, 109 Alexander Drive, Mail Drop B105-03, Research Triangle Park, NC, 27711, USA.
Environmental Public Health Division, National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, Research Triangle Park, NC, 27711, USA.
Cell Biol Toxicol. 2023 Oct;39(5):2311-2329. doi: 10.1007/s10565-022-09720-6. Epub 2022 Jul 25.
In dose-response and structure-activity studies, human hepatic HepG2 cells were exposed for 3 days to nano Cu, nano CuO or CuCl (ions) at doses between 0.1 and 30 ug/ml (approximately the no observable adverse effect level to a high degree of cytotoxicity). Various biochemical parameters were then evaluated to study cytotoxicity, cell growth, hepatic function, and oxidative stress. With nano Cu and nano CuO, few indications of cytotoxicity were observed between 0.1 and 3 ug/ml. In respect to dose, lactate dehydrogenase and aspartate transaminase were the most sensitive cytotoxicity parameters. The next most responsive parameters were alanine aminotransferase, glutathione reductase, glucose 6-phosphate dehydrogenase, and protein concentration. The medium responsive parameters were superoxide dismutase, gamma glutamyltranspeptidase, total bilirubin, and microalbumin. The parameters glutathione peroxidase, glutathione reductase, and protein were all altered by nano Cu and nano CuO but not by CuCl exposures. Our chief observations were (1) significant decreases in glucose 6-phosphate dehydrogenase and glutathione reductase was observed at doses below the doses that show high cytotoxicity, (2) even high cytotoxicity did not induce large changes in some study parameters (e.g., alkaline phosphatase, catalase, microalbumin, total bilirubin, thioredoxin reductase, and triglycerides), (3) even though many significant biochemical effects happen only at doses showing varying degrees of cytotoxicity, it was not clear that cytotoxicity alone caused all of the observed significant biochemical effects, and (4) the decreased glucose 6-phosphate dehydrogenase and glutathione reductase support the view that oxidative stress is a main toxicity pathway of CuCl and Cu-containing nanomaterials.
在剂量反应和结构活性研究中，用人肝 HepG2 细胞在 0.1 至 30ug/ml（约为高细胞毒性的无可见不良作用水平）的剂量下暴露于纳米 Cu、纳米 CuO 或 CuCl（离子）3 天。然后评估各种生化参数以研究细胞毒性、细胞生长、肝功能和氧化应激。对于纳米 Cu 和纳米 CuO，在 0.1 至 3ug/ml 之间观察到很少的细胞毒性迹象。就剂量而言，乳酸脱氢酶和天冬氨酸转氨酶是最敏感的细胞毒性参数。其次最敏感的参数是丙氨酸转氨酶、谷胱甘肽还原酶、葡萄糖 6-磷酸脱氢酶和蛋白质浓度。其次敏感的参数是超氧化物歧化酶、γ-谷氨酰转肽酶、总胆红素和微量白蛋白。谷胱甘肽过氧化物酶、谷胱甘肽还原酶和蛋白质参数都被纳米 Cu 和纳米 CuO 改变，但 CuCl 暴露没有改变。我们的主要观察结果是：（1）在表现出高细胞毒性的剂量以下观察到葡萄糖 6-磷酸脱氢酶和谷胱甘肽还原酶显著降低，（2）即使高细胞毒性也不会在一些研究参数中引起大的变化（例如碱性磷酸酶、过氧化氢酶、微量白蛋白、总胆红素、硫氧还蛋白还原酶和甘油三酯），（3）尽管许多显著的生化效应仅在表现出不同程度细胞毒性的剂量下发生，但尚不清楚细胞毒性单独导致了所有观察到的显著生化效应，（4）葡萄糖 6-磷酸脱氢酶和谷胱甘肽还原酶的降低支持了氧化应激是 CuCl 和含 Cu 纳米材料的主要毒性途径的观点。
