Diethylnitrosamine Induced Oxidative Stress and Morphological Changes in The Cerebellum of Wistar Rat
Diethylnitrosamine (DEN), a known toxin and potent carcinogen induces oxidative stress by generation of free radicals resulting in cellular injury through its metabolized end product. There is dearth of information on DEN neurotoxicity. This study therefore, evaluated the oxidative damage and morphological changes induced by DEN in the cerebellum of Wistar rat. Twenty male Wistar rats weighing between 110 and 120 g were divided into two groups (n=10). Group I rats received distilled water and served as the control group, while group II rats received 25 mg/kg body weight of DEN i.p. twice weekly for 12 weeks. At the end of the administration, the rats from both groups were weighed and killed. The brains were weighed and the cerebella dissected out, some preserved in phosphate buffered saline for oxidative stress and antioxidant markers, while others fixed in 10% formol-saline for histological and immunohistochemical (Glial fibrillary acidic protein, GFAP) studies, for cerebellar cytoarchitecture and astrocytes population, respectively. Data were analyzed employing the unpaired student’s t-test at p<0.05. There was a significant decrease in weight gain in the DEN-treated group, increased lipid peroxidation, decreased glutathione levels, superoxide dismutase activity and compared with the control rats at p<0.05. Histological and histomorphometric evaluation of the cerebellar cortex of DEN-treated rats showed pyknosis of the Purkinje cells with chromatolysis, significant reduction in the diameter/size and loss of the Purkinje cells compared with the control rats. Immunohistochemically, there was increased population of astrocytes in the DEN-treated rats compared with the control rats. The results of the study shows that prolonged exposure of rats to diethylnitrosamine induced oxidative stress with morphological alterations in the cerebellum and thus increasing the literature of diethylnitrosamine neurotoxicity
Archer, M. C. (1989). Mechanisms of action of N-nitroso compounds. Cancer Survey 8 (2) :241–250.
Attwell, P. J., Cooke, S. F. and Yeo, C. H. (2002). Cerebellar function in consolidation of a motor memory. Neuron 34: 1011-1020.
Bansal, A. K., Bansal, M., Soni, G. and Bhatnagar, D. (2005). Protective role of Vitamin E pretreatment on N-nitrosodiethylamine induced oxidative stress in rat liver. Chemico-Biological Interaction 156: 101–111.
Bendong, C., Mingliang, N. and Guangshun, Y. (2012). Effect of paeonol on antioxidant and immune regulatory activity in HCC Rats. Molecules 17: 4672–4683.
Brown, J. L. (1999). N-Nitrosamines. Occupational Medicine. 14: 839–848.
Chuang, C. H. and Hu, M. L. (2006). Synergistic DNA damage and lipid peroxidation in cultured human white blood cells exposed to 4-(methyl-nitrosamino)-1-(3-pyridyl)-1-butanone and ultraviolet A. Environmental Molecular Mutagenesis. 47: 73–81.
De la Monte, S. M. and Tong, M. (2009). Mechanisms Of Nitrosamine–Mediated Neurodegeneration: Potential Relevance To Sporadic Alzheimer’s Disease Journal of Alzheimers Disease. 17 (4): 817–825.
Elguindya, N. M., Yacouta, G. A. and El Azabb, E. F. (2018). Amelioration of DENA-induced oxidative stress in rat kidney and brain by the essential oil of Elettaria cardamomum. Beni-Suef University Journal of Basic and Applied Sciences 7 (3): 299-305.
Eng, L. F., Ghirnikar, R. S. and Lee, Y. L. (2000) Glial fibrillary acidic protein: GFAP-thirty-one years (1969–2000). Neurochemical Research 25:1439-1451.
Fonnum, F. and Lock, E. A. (2000). Cerebellum as a target for toxic substances. Toxicol. Letters. 15: 112-113, 9- 16.
Gray, C., Perciavalle, V. and Poppele, R. (1993). Sensory responses to passive hindlimb joint rotation in the cerebellar cortex of the cat. Brain Research 622: 280-284.
Gray, J. I., Skrypec, D. J., Mandagere, A. K., Booren, A. M. and Pearson, A. M. (1984). Further factors influencing N-nitrosamine formation in bacon. International Agency for Research on Cancer (IARC) Scientific Publications 301–309.
Gudi, V., Moharregh-Khiabani, D., Skripuletz, T., Koutsoudaki, P.N., Kotsiari, A. and Skul-jec, J. (2009). Regional differences between grey and white matter in cuprizone induced demyelination. Brain Research 1283: 127-138.
Ha, W. S., Kim, C. K., Song, S. K. and Kang, C. B. (2001). Study on mechanism of multistep hepatotumorigenesis in rat: Development of hepatotumorigenesis. Journal of Veterinary Sciences 2: 53-58.
Jakszyn, P. and Gonzalez, C. A. (2006). Nitrosamine and related food intake and gastric and oesophageal cancer risk: a systematic review of the epidemiological evidence. World Journal of Gastroenterology 12 (27): 4296–303.
Jollow, D. J., Mitchell, J. R., Zamppaglione, Z. and Gillette, J. R. (1974). Bromobenzene induced liver necrosis. Protective role of glutathione and evidence for 3,4-bromobenzene oxide as the hepatotoxic metabolites. Pharmacology 11: 151-157.
Kadasa, N. M, Abdallah, H., Afifi, M. and Gowayed, S. (2015). Hepatoprotective
effects of curcumin against diethyl nitrosamine induced hepatotoxicity in albino rats. Asian Pacific Journal of Cancer Prevention 16: 103-108.
Kettenmann, H. and Verkhratsky, A. (2011). Neuroglia - Living Nerve Glue. Fortschritte der Neurologie und Psychiatrie 79:588-597.
Kim, D., Jeond, S. and Lee, C. (2003). Antioxidant capacity of phenolic phytochemicals from various cultivars of plums. Food Chemistry 81: 321-326.
Liao, D. J., Blanck, A., Eneroth, P., Gustafsson, J. A. and Hällström, I. P. (2001).
Diethylnitrosamine causes pituitary damage, disturbs hormone levels and reduces
sexual dimorphism of certain liver functions in the rat. Environmental Health Perspectives 109: 943–947.
Lopez-Novoa, J. M., Quiros, Y., Vicente, L., Morales, A. I. and Lopez-Hernandez, F. J. (2011). New insights into the mechanism of aminoglycoside nephrotoxicity: an integrative point of view. Kidney International 79: 33–45.
Maier, C. M. and Chan, P. H. (2002). Role of superoxide dismutases in oxidative damage and neurodegenerative disorders. The Neuroscientist 8: 323–334.
Manto, M. (2012). Toxic agents causing cerebellar ataxias. Handbook of Clinical Neurology 103: 201-213.
Misra, H. and Fridovich, I. (1972). The role of superoxide anion in the auto-oxidation of epinephrine and a simple assay for superoxide dismutase. Journal of Biological Chemistry 217 (10): 3170-3175.
Mittal, G., Brar, A. P. and Soni, G. (2006). Impact of hypercholesterolemia on toxicity of N-nitrosodiethylamine: biochemical and histopathological effects. Pharmacological Reports 58: 413-419.
Moustafa, M. E., Mohamed, A. M. and Thabet, M. N. (2017). Gallium Nanoparticle-Mediated Reduction of Brain Specific Serine Protease-4 in an Experimental Metastatic Cancer Model. Asian Pacific Journal Cancer Prevention 18 (4): 895–903.
Ota, Y., Zanetti, A. T., Hallock, R. M. (2013). The role of astrocytes in the regulation of synaptic plasticity and memory formation. Neural Plasticity 2013: 1-11.
Park, D. H., Shin, J. W., Park, S. K., Seo, J. N., Li, L., Jang, J. J. and Lee, M. J. (2009). Diethylnitrosamine (DEN) induces irreversible hepatocellular carcinogenesis through overexpression of G1/S-phase regulatory proteins in rat. Toxicology Letters 15: 191(2-3): 321-326.
Poirier, M. C. and Beland, F. A. (1994). Deoxyribonucleic acid adduct measurements and tumor incidence during chronic carcinogen exposure in rodents. Environmental Health Perspectives 102(suppl 6):161–165.
Pylypiw, H. M., Zubroff, J. R., Magee, P. N. and Harrington, G. W. (1984). The metabolism of N-nitrosomethylaniline. Journal of Cancer Research and Clinical Oncology.108: 66-70.
Rakic, P. and Sidman, R. L. (1970). Histogenesis of cortical layers in human cerebellum particularly the lamina dissecans. Journal of Comparative Neurology 139: 473.
Rotruck, J. T., Pope, A. L., Ganther, H. E., Swanson, A. B., Hafeman, D. G. and Hoekstra, W. G. (1973). Biochemical role as a component of glutathione peroxidase. Science, 179: 588-590.
Schuller, H. M. (1992. Nitrosamine-induced lung carcinogenesis and Ca2+/calmodulin antagonists. Cancer Researcg 52 (suppl 9): 2723s-2726s.
Sheweita, A. S., El-Bendery, H. A. and Mostafa, H. M. (2014). Novel Study on N-Nitrosamines as Risk Factors of Cardiovascular Diseases. Biomedical Research International 2014: 817019.
Sheweita, S. A., Mousa, N. and Al-Masry, H. M. (2008). N-Nitrosodimethylamine changes the expression of glutathione S-transferase in the liver of male mice: the role of antioxidants. Journal of Biochemistry and Molecular Toxicology 22 (6): 389–395.
Snell, R. S. (2001). Clinical Neuroanatomy for Medical students, 5th edition. Lippincott. William and Wilkins pp 228.
Sofroniew, M. V. and Vinters, H. V. (2010). Astrocytes: biology and pathology Acta Neuropathology 119: 7-35.
Şovrea, A. S. and Boşca, A. B. (2013). Astrocytes reassessment - an evolving concept part one: embryology, biology, morphology and reactivity. Journal of Molecular Psychiatry 1: 18.
Sivaramakrishnan. V., Shilpa, P. N., Praveen Kumar, V. R. and Niranjali Devaraj, S. (2008). Attenuation of N-nitrosodiethylamine-induced hepatocellular carcinogenesis by a novel flavonol-Morin. Chemico-Biological Interaction 171: 79-88.
Tricker, A. R., Perkins, M. J., Massey, R. C. and McWeeny, D. J. (1985). Some nitrosoamino acids in bacon adipose tissue and their contribution to the total N-nitroso compound concentration. Z Lebensm Unters Forsch 180:379-383.
Varshney, R. and Kale, R. (1990). Effect of calmodulin antagonist on radiation induced lipid peroxidation in microsomes. International Journal Radiation Biology 58: 733-743.
Verna, L., Whysner, J. and Williams, G. M. (1996). N-nitrosodiethylamine mechanistic data and risk asseessment: bioactivation, DNA-adduct formation, mutagenicity, and tumor initiation. Pharmacology and Therapeutics 71:57-81.
Vitaglione, P., Morisco, F., Caporaso, N. and Fogliano, V. (2004). Dietary antioxidant compounds and liver health. Critical Review Food Science and Nutrition 44 (7-8): 575-86.
West, I. C. (2000). Radicals and oxidative stress in diabetes. Diabetic Medicine 17: 171-180.
Zaaraoui, W., Deloire, M., Merle, M., Girard, C., Raffard, G. and Biran, M. (2008). Monitoring demyelination and remyelination by magnetization transfer imaging in the mouse brain at 9.4 T. MAGMA 21: 357-362.