Abstract
Molybdenum (Mo) is an essential trace element for animals and humans. However, the high dietary intake of Mo leads to disease conditions in heavy metal pollution areas. To the best of our knowledge, the effect of high levels of Mo on the apoptosis of hepatocyte in goats has not been investigated. Therefore, the aim of the present in vivo study was to investigate the impact of Mo on mitochondrial oxidative stress and apoptosis genes in the liver using real-time quantitative polymerase chain reaction (RT-qPCR) and transmission electron microscopy, respectively. Thirty-six healthy goats were randomly divided into three groups: two groups treated with ammonium molybdate [(NH4)6·Mo7O24·H2O] at 15 and 45 mg Mo kg−1 BW, respectively, and a control group without treatment. Liver samples were collected from individual goats at different time intervals. The levels of oxidative stress in the mitochondrial membrane and expression of liver-related apoptosis genes, including Bcl-2, Cyt c, caspase-3, and Smac, were examined. The results demonstrated that the levels of superoxide dismutase (SOD) and catalase (CAT) expression were significantly down-regulated in liver cells, whereas malondialdehyde (MDA), nitric oxide (NO), and total nitric oxide synthase (T-NOS) expression was up-regulated (P < 0.01). The expression of Smac, Cyt c, and caspase-3 was significantly up-regulated, whereas Bcl-2 expression was down-regulated in liver cells (P < 0.01). In addition, histopathological examination revealed varying degrees of vacuolization, irregularity, nuclear fission, and mitochondrial swelling and high-density electrons in the cytoplasm of hepatocytes in groups treated with 15 and 45 mg Mo kg−1 BW. Thus, these results suggested that high molybdenum induced hepatocyte apoptosis and might involve a mitochondrial pathway.
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References
Schwarz G, Mendel RR, Ribbe MW (2009) Molybdenum cofactors, enzymes and pathways. Nature 460:839–47
Nordberg G (2007) Handbook on the toxicology of metals, 3rd edn. Academic Press, Amsterdam
Tejada-Jimenez M, Galvan A, Fernandez E, Llamas A (2009) Homeostasis of the micronutrients Ni, Mo and Cl with specific biochemical functions. Curr Opin Plant Biol 12:358–63
Wei Z, Yin M, Zhang X, Hong F, Li B, Tao Y, Zhao G, Yan C (2001) Rare earth elements in naturally grown fern Dicranopteris linearis in relation to their variation in soils in south-Jiangxi region (southern China). Environ Pollut 114:345–55
Barceloux DG (1999) Molybdenum. J Toxicol Clin Toxicol 37:231–7
Pitt MA (1976) Molybdenum toxicity: interactions between copper, molybdenum and sulphate. Agents Actions 6:758–69
Vyskocil A, Viau C (1999) Assessment of molybdenum toxicity in humans. J Appl Toxicol 19:185–92
Kang J, Pervaiz S (2012) Mitochondria: redox metabolism and dysfunction. Biochem Res Int 2012:896751
Zaidi A, Fernandes D, Bean JL, Michaelis ML (2009) Effects of paraquat-induced oxidative stress on the neuronal plasma membrane Ca(2+)-ATPase. Free Radic Biol Med 47:1507–14
Su M, Mei Y, Sinha S (2013) Role of the crosstalk between autophagy and apoptosis in cancer. J Oncol 2013:102735
Naziroglu M, Karaoglu A, Aksoy AO (2004) Selenium and high dose vitamin E administration protects cisplatin-induced oxidative damage to renal, liver and lens tissues in rats. Toxicology 195:221–30
Sreedhar R, Arumugam S, Thandavarayan RA, Giridharan VV, Karuppagounder V, Pitchaimani V, Afrin R, Miyashita S, Nomoto M, Harima M, Gurusamy N, Suzuki K, Watanabe K (2015) Myocardial 14-3-3eta protein protects against mitochondria mediated apoptosis. Cell Signal.
Green DR, Reed JC (1998) Mitochondria and apoptosis. Science 281:1309–12
National Research Council (1981) Nutrient requirements of goats: Angora, dairy, and meat goats in temperate and tropical countries. National Academy Press, Washington, DC
Tang Z, Iqbal M, Cawthon D, Bottje WG (2002) Heart and breast muscle mitochondrial dysfunction in pulmonary hypertension syndrome in broilers (Gallus domesticus). Comp Biochem Physiol A Mol Integr Physiol 132:527–40
Liu XF, Zhang LM, Guan HN, Zhang ZW, Xu SW (2013) Effects of oxidative stress on apoptosis in manganese-induced testicular toxicity in cocks. Food Chem Toxicol 60:168–76
Mertz W, Underwood EJ (1986) Trace elements in human and animal nutrition, 5th edn. Academic Press, Orlando
Walter M (2004) Clinical signs in cattle grazing high molybdenum forage. J Range Manage 57:269–74
Liu W, Zhao H, Wang Y, Jiang C, Xia P, Gu J, Liu X, Bian J, Yuan Y, Liu Z (2014) Calcium-calmodulin signaling elicits mitochondrial dysfunction and the release of cytochrome c during cadmium-induced apoptosis in primary osteoblasts. Toxicol Lett 224:1–6
Liu S, Xu FP, Yang ZJ, Li M, Min YH, Li S (2014) Cadmium-induced injury and the ameliorative effects of selenium on chicken splenic lymphocytes: mechanisms of oxidative stress and apoptosis. Biol Trace Elem Res 160:340–51
Lu X, Wang C, Liu B (2015) The role of Cu/Zn-SOD and Mn-SOD in the immune response to oxidative stress and pathogen challenge in the clam Meretrix meretrix. Fish Shellfish Immunol 42:58–65
Zhang W, Zhang Y, Zhang SW, Song XZ, Jia ZH, Wang RL (2012) Effect of different levels of copper and molybdenum supplements on serum lipid profiles and antioxidant status in cashmere goats. Biol Trace Elem Res 148:309–15
Rousseaux CG, Haschek WM, Wallig MA (2002) Handbook of toxicologic pathology, 2nd edn. Academic Press, San Diego
Frank A, Danielsson R, Jones B (2000) The ‘mysterious’ disease in Swedish moose. Concentrations of trace elements in liver and kidneys and clinical chemistry. Comparison with experimental molybdenosis and copper deficiency in the goat. Sci Total Environ 249:107–22
Frank A (2004) A review of the “mysterious” wasting disease in Swedish moose (Alces alces L.) related to molybdenosis and disturbances in copper metabolism. Biol Trace Elem Res 102:143–59
O’Connor JM (2001) Trace elements and DNA damage. Biochem Soc Trans 29:354–7
Park MY, Jeong YJ, Kang GC, Kim MH, Kim SH, Chung HJ, Jung JY, Kim WJ (2014) Nitric oxide-induced apoptosis of human dental pulp cells is mediated by the mitochondria-dependent pathway. Korean J Physiol Pharmacol 18:25–32
Chang KC, Hsu CC, Liu SH, Su CC, Yen CC, Lee MJ, Chen KL, Ho TJ, Hung DZ, Wu CC, Lu TH, Su YC, Chen YW, Huang CF (2013) Cadmium induces apoptosis in pancreatic beta-cells through a mitochondria-dependent pathway: the role of oxidative stress-mediated c-Jun N-terminal kinase activation. PLoS ONE 8, e54374
Liu CM, Ma JQ, Sun YZ (2012) Puerarin protects the rat liver against oxidative stress-mediated DNA damage and apoptosis induced by lead. Exp Toxicol Pathol 64:575–82
Zhao S, Xiong Z, Mao X, Meng D, Lei Q, Li Y, Deng P, Chen M, Tu M, Lu X, Yang G, He G (2013) Atmospheric pressure room temperature plasma jets facilitate oxidative and nitrative stress and lead to endoplasmic reticulum stress dependent apoptosis in HepG2 cells. PLoS ONE 8, e73665
Liu X, Zuo N, Guan H, Han C, Xu SW (2013) Manganese-induced effects on cerebral trace element and nitric oxide of Hyline cocks. Biol Trace Elem Res 154:202–9
Xiao J, Cui HM, Yang F, Peng X, Cui Y (2011) Effect of dietary high molybdenum on the cell cycle and apoptosis of kidney in broilers. Biol Trace Elem Res 142:523–31
Yang F, Cui H, Xiao J, Peng X, Deng J, Zuo Z (2011) Increased apoptotic lymphocyte population in the spleen of young chickens fed on diets high in molybdenum. Biol Trace Elem Res 140:308–16
Kluck RM, Bossy-Wetzel E, Green DR, Newmeyer DD (1997) The release of cytochrome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis. Science 275:1132–6
Tsujimoto Y (2002) Bcl-2 family of proteins: life-or-death switch in mitochondria. Biosci Rep 22:47–58
Pulido MD, Parrish AR (2003) Metal-induced apoptosis: mechanisms. Mutat Res 533:227–41
Nicholson DW, Thornberry NA (1997) Caspases: killer proteases. Trends Biochem Sci 22:299–306
Larsen BD, Rampalli S, Burns LE, Brunette S, Dilworth FJ, Megeney LA (2010) Caspase 3/caspase-activated DNase promote cell differentiation by inducing DNA strand breaks. Proc Natl Acad Sci U S A 107:4230–5
Acknowledgments
The research was financially supported through grants from the Program of the National Nature Science Foundation (no. 31101863, Beijing, People’s Republic of China), the Training Plan for Young Scientists of Jiangxi province (no. 2014BCB23040, Nanchang, People’s Republic of China), and the Educational Departmental Science Foundation of Jiangxi province (No.GJJ14294, Nanchang, People’s Republic of China) to Huabin Cao.
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Yu Zhuang and Ping Liu contributed equally to this work.
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Zhuang, Y., Liu, P., Wang, L. et al. Mitochondrial oxidative stress-induced hepatocyte apoptosis reflects increased molybdenum intake in caprine. Biol Trace Elem Res 170, 106–114 (2016). https://doi.org/10.1007/s12011-015-0450-0
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DOI: https://doi.org/10.1007/s12011-015-0450-0