Facultad de Ciencias Médicas, Instituto de Investigaciones Bioquímicas de La Plata, CCT La Plata, CONICET-UNLP, Cátedra de Bioquímica y Biología Molecular, Universidad Nacional de La Plata, La Plata, Argentina.
Hum Exp Toxicol. 2013 Mar;32(3):299-315. doi: 10.1177/0960327112456313. Epub 2012 Aug 16.
We investigated the effect of copper (Cu) overload (20-160 µM/24 h) in two cell lines of human hepatic (HepG2) and pulmonary (A-549) origin by determining lipid and protein damage and the response of the antioxidant defence system. A-549 cells were more sensitive to Cu overload than HepG2 cells. A marked increase was observed in both the cell lines in the nitrate plus nitrite concentration, protein carbonyls and thiobarbituric acid reactive substances (TBARS). The TBARS increase was consistent with an increment in saturated fatty acids at the expense of polyunsaturated acids in a Cu concentration-dependent fashion. Antioxidant enzymes were stimulated by Cu overload. Superoxide dismutase activity increased significantly in both the cell lines, with greater increases in HepG2 than in A-549 cells. A marked increase in ceruloplasmin and metallothionein content in both the cell types was also observed. Dose-dependent decreases in α-tocopherol and ferric reducing ability were observed. Total glutathione content was lower in A-549 cells and higher in HepG2. Calpain and caspase-3 were differentially activated in a dose-dependent manner under copper-induced reactive oxygen species production. We conclude that Cu exposure of human lung- and liver-derived cells should be considered a reliable experimental system for detailed study of mechanism/mechanisms by which Cu overload exerts its deleterious effects.
我们通过测定脂质和蛋白质损伤以及抗氧化防御系统的反应，研究了铜（Cu）过载（20-160 μM/24 h）对两种人源肝（HepG2）和肺（A-549）细胞系的影响。A-549 细胞比 HepG2 细胞对铜过载更敏感。在两种细胞系中，硝酸盐加亚硝酸盐浓度、蛋白质羰基和硫代巴比妥酸反应物质（TBARS）均明显增加。TBARS 的增加与饱和脂肪酸的增加一致，而多不饱和脂肪酸则以 Cu 浓度依赖的方式减少。抗氧化酶被铜过载所刺激。超氧化物歧化酶活性在两种细胞系中均显著增加，HepG2 细胞的增加大于 A-549 细胞。两种细胞类型的铜蓝蛋白和金属硫蛋白含量也明显增加。α-生育酚和铁还原能力呈剂量依赖性下降。A-549 细胞中的总谷胱甘肽含量较低，而 HepG2 细胞中的含量较高。钙蛋白酶和半胱天冬酶-3 在铜诱导的活性氧产生下以剂量依赖性方式被不同程度地激活。我们得出结论，铜暴露于人肺和肝源性细胞应被视为详细研究铜过载产生有害影响的机制/机制的可靠实验系统。
