Cadmium-induced apoptosis through the mitochondrial pathway in rainbow trout hepatocytes: involvement of oxidative stress

• Published in: Aquatic toxicology Vol. 69; no. 3; pp. 247 - 258
• Main Authors: Risso-de Faverney, C, Orsini, N, de Sousa, G, Rahmani, R
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 25.08.2004; Elsevier Science; Elsevier
• Subjects: 2,2,6,6-tetramethylpiperidinyl-1-oxyl; Acetylcysteine - pharmacology; Agnatha. Pisces; Animal, plant and microbial ecology; Animals; antioxidants; Antioxidants - pharmacology; Apoptosis; Apoptosis - drug effects; Applied ecology; bcl-2-Associated X Protein; Biological and medical sciences; Blotting, Western; Cadmium; Cadmium - toxicity; caspase; Caspases - metabolism; Cyclic N-Oxides - pharmacology; cytochrome c; Cytochromes c - metabolism; cytosol; Dose-Response Relationship, Drug; Ecotoxicology, biological effects of pollution; Effects of pollution and side effects of pesticides on vertebrates; electrophoresis; enzyme activity; Fish; Freshwater; Fundamental and applied biological sciences. Psychology; gene expression; hatchery-reared fish; heavy metals; Hepatocytes; Hepatocytes - metabolism; Life Sciences; Mitochondria; Mitochondria - metabolism; N-acetylcysteine; Oncorhynchus mykiss; Oncorhynchus mykiss - metabolism; Oxidative stress; Oxidative Stress - drug effects; proteins; Proto-Oncogene Proteins - metabolism; Proto-Oncogene Proteins c-bcl-2; reactive oxygen species; Reactive Oxygen Species - metabolism; Statistics, Nonparametric; Time Factors; trout; water pollution; Western blot analysis; Cadmium; Oxidative stress; Fish; Mitochondria; Hepatocytes; Apoptosis; Salmonidae; Ecotoxicology; Digestive system; Liver; Oncorhynchus mykiss; Heavy metal; Aquatic environment; Vertebrata; Hepatocyte; Pisces; APOPTOSIS; HEPATOCYTES; OXIDATIVE STRESS; CADMIUM; MITOCHONDRIA; FISH; TOXICOLOGIE
• ISSN: 0166-445X; 1879-1514
• DOI: 10.1016/j.aquatox.2004.05.011

Cadmium (Cd) induces oxidative stress and apoptosis in trout hepatocytes. We therefore investigated the involvement of the mitochondrial pathway in the initiation of apoptosis and the possible role of oxidative stress in that process. This study demonstrates that hepatocyte exposure to Cd (2, 5 and 10 μM) triggers significant caspase-3, but also caspase-8 and -9 activation in a dose-dependent manner. Western-blot analysis of hepatocyte mitochondrial and cytosolic fractions revealed that cytochrome c (Cyt c) was released in the cytosol in a dose-dependent manner, whereas the pro-apoptotic protein Bax was redistributed to mitochondria after 24 and 48 h exposure. We also found that the expression of anti-apoptotic protein Bcl-xL, known to be regulated under mild oxidative stress to protect cells from apoptosis, did not change after 3 and 6 h exposure to Cd, then increased after 24 and 48 h exposure to 10 μM Cd. In the second part of this work, two antioxidant agents, 2,2,6,6-tetramethylpiperidinyl-1-oxyl (TEMPO) (100 μM) and N-acetylcysteine (NAC, 100 μM) were used to determine the involvement of reactive oxygen species (ROS) in Cd-induced apoptosis. Simultaneously exposing trout hepatocytes to Cd and TEMPO or NAC significantly reduced caspase-3 activation after 48 h and had a suppressive effect on caspase-8 and -9 also, mostly after 24 h. Lastly, the presence of either one of these antioxidants in the treatment medium also attenuated Cd-induced Cyt c release in cytosol and the level of Bax in the mitochondria after 24 and 48 h, while high Bcl-xL expression was observed. Taken together, these data clearly evidenced the key role of mitochondria in the cascade of events leading to trout hepatocyte apoptosis in response to Cd and the relationship that exists between oxidative stress and cell death.