Effects of ammonia on apoptosis and oxidative stress in bovine mammary epithelial cells

• Published in: Mutagenesis Vol. 33; no. 4; pp. 291 - 299
• Main Authors: Wang, Fengge, Chen, Shuxiong, Jiang, Yanwen, Zhao, Yun, Sun, Liting, Zheng, Biaobiao, Chen, Lu, Liu, Zhuo, Zheng, Xue, Yi, Kangle, Li, Chunjin, Zhou, Xu
• Format: Journal Article
• Language: English
• Published: UK Oxford University Press 11.10.2018
• Subjects: Ammonia - pharmacology; Animals; Apoptosis - drug effects; Calcium - metabolism; Cattle; Cell Survival - drug effects; Cells, Cultured; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Mammary Glands, Animal - drug effects; Mammary Glands, Animal - metabolism; Membrane Potential, Mitochondrial - drug effects; Oxidative Stress - drug effects; Phosphorylation - drug effects; Reactive Oxygen Species - metabolism; Signal Transduction - drug effects; Tumor Suppressor Protein p53 - metabolism
• ISSN: 0267-8357; 1464-3804
• DOI: 10.1093/mutage/gey023

Abstract Ammonia, produced mainly from the deamination of amino acids and glutamine, is one of the major toxic components in blood and tissues that may affect bovine health. However, the physiological and pathological roles of ammonia in the mammary glands are not understood clearly. In the present study, the bovine mammary epithelial cell line (MAC-T) was utilised as an in vitro model to determine the effects of ammonia on bovine mammary gland. We demonstrated that ammonia stimulated the production of intracellular reactive oxygen species, decreased mitochondrial membrane potential, interrupted intracellular calcium ion (Ca2+) homeostasis and induced cell apoptosis. Ammonia also significantly reduced cell viability and increased the proportion of apoptotic cells through enhancing the level of p53 phosphorylation and increasing the expressions of BAX, caspase 8, caspase 9, caspase 3. Interestingly, bumetanide, a specific Na+ K+ 2Cl−-cotransporter inhibitor, dramatically abolished the damaging effects of ammonia on the cells. These data suggest that ammonia exposure induces apoptosis in bovine mammary epithelial cells via activation of the p53 pathway and the mitochondrial apoptotic pathway, and that these effects involved the Na+ K+ 2Cl−-cotransporter.