Effects of Acute Alcohol Intoxication on Testicular Dna Stability, Gene Expression of Cytochromes CYP3A and CYP2E1, and Serum Pool of Free Amino Acids in Rats

Authors

DOI:

https://doi.org/10.20535/ibb.2024.8.4.317136

Keywords:

acute alcohol intoxication, DNA fragmentation, gene expression, CYP450, pool of free amino acids, testis

Abstract

Background. Alcohol's toxic effects on the organism is a long-known medical problem. Alcohol's damaging effect is the end result of the complex interplay between ethanol metabolism, inflammation and innate immunity. Previously, we studied the long-term consequences of chronic alcoholism and demonstrated that especially profound changes were in testes on the level of proteome and genome.

Objective. This work aimed to study short-term acute alcohol intoxication (AAI) effects for rat testis DNA fragmentation, cytochromes CYP3A and CYP2E1 genes expression, and serum pool of free amino acids in rats.

Methods. Wistar albino male rats were divided into 2 groups (8 animals in each group): 1 – Control (intact rats), and 2 – AAI (rats with short-term acute alcohol intoxication). AAI was induced by repeated administration per os 40% ethanol solution in a dose 7 ml/kg body weight, for 7 days. Contents of amino acids in serum, testes mRNA CYP2E1 and CYP3A2 expression, and DNA fragmentation were evaluated.

Results. In our experiments, the development of acute alcohol intoxication (AAI) led to increased DNA fragmentation processes in the testes of adult rats compared to the control group. Additionally, in the serum of ethanol-treated rats, the levels of histidine increased by 1.67 times and glutamine by 1.13 times in correlation with this pathology. Conversely, the levels of valine, phenylalanine, as well as non-essential and essential amino acids, decreased. Furthermore, there was a statistically significant increase in the expression of CYP2E1 and CYP3A2 genes in rat testes under the conditions of AAI.

Conclusions. In conclusion, investigation of rats' short-term alcohol administration effects permitted us to obtain the picture of complex metabolomic changes at the different levels. The main outcome of rats short-term ethanol administration in our experiments seems to be to some extent similar to changes described for rats with chronic alcohol consumption. Our results demonstrated profound changes in testes affecting the state of the genome, transcription processes and the exchange of amino acids and proteins. We suggest that the revealed testicular metabolic disorders could have negative implications on cellular regulation of spermatogenesis even under short-term ethanol exposure.

References

Campollo O. Alcohol and the liver: The return of the prodigal son. Ann Hepatol. 2019;18(1):6-10. DOI: 10.5604/01.3001.0012.7854

O'Shea RS, Dasarathy S, McCullough AJ; Practice Guideline Committee of the American Association for the Study of Liver Diseases; Practice Parameters Committee of the American College of Gastroenterology. Alcoholic liver disease. Hepatology. 2010 Jan;51(1):307-28. DOI: 10.1002/hep.23258

Michalak A, Lach T, Cichoż-Lach H. Oxidative stress-A key player in the course of alcohol-related liver disease. J Clin Med. 2021 Jul 6;10(14):3011. DOI: 10.3390/jcm10143011

Costa PA, Poli JH, Sperotto ND, Moura DJ, Saffi J, Nin MS, et al. Brain DNA damage and behavioral changes after repeated intermittent acute ethanol withdrawal by young rats. Psychopharmacology (Berl). 2015 Oct;232(19):3623-36. DOI: 10.1007/s00213-015-4015-x

Jilek JL, Sant KE, Cho KH, Reed MS, Pohl J, Hansen JM, Harris C. Ethanol attenuates histiotrophic nutrition pathways and alters the intracellular redox environment and thiol proteome during rat organogenesis. Toxicol Sci. 2015 Oct;147(2):475-89. DOI: 10.1093/toxsci/kfv145

Sadeghzadeh M, Shirpoor A, Naderi R, Kheradmand F, Gharalari FH, Samadi M, et al. Long-term ethanol consumption promotes changes in β-defensin isoform gene expression and induces structural changes and oxidative DNA damage to the epididymis of rats. Mol Reprod Dev. 2019 Jun;86(6):624-31. DOI: 10.1002/mrd.23138

Naseer MI, Ullah I, Narasimhan ML, Lee HY, Bressan RA, Yoon GH, et al. Neuroprotective effect of osmotin against ethanol-induced apoptotic neurodegeneration in the developing rat brain. Cell Death Dis. 2014;5(3):e1150. DOI: 10.1038/cddis.2014.53

Chayanupatkul M, Liangpunsakul S. Alcoholic hepatitis: a comprehensive review of pathogenesis and treatment. World J Gastroenterol. 2014 May 28;20(20):6279-86. DOI: 10.3748/wjg.v20.i20.6279

Gao B, Bataller R. Alcoholic liver disease: pathogenesis and new therapeutic targets. Gastroenterology. 2011 Nov;141(5):1572-85. DOI: 10.1053/j.gastro.2011.09.002

Laso FJ, Lapeña P, Madruga JI, San Miguel JF, Orfao A, Iglesias MC, et al. Alterations in tumor necrosis factor-alpha, interferon-gamma, and interleukin-6 production by natural killer cell-enriched peripheral blood mononuclear cells in chronic alcoholism: relationship with liver disease and ethanol intake. Alcohol Clin Exp Res. 1997 Oct;21(7):1226-31.

Spahr L, Giostra E, Frossard JL, Bresson-Hadni S, Rubbia-Brandt L, Hadengue A. Soluble TNF-R1, but not tumor necrosis factor alpha, predicts the 3-month mortality in patients with alcoholic hepatitis. J Hepatol. 2004 Aug;41(2):229-34. DOI: 10.1016/j.jhep.2004.04.028

Stewart S, Jones D, Day CP. Alcoholic liver disease: new insights into mechanisms and preventative strategies. Trends Mol Med. 2001 Sep;7(9):408-13. DOI: 10.1016/s1471-4914(01)02096-2

McClain CJ, Shedlofsky S, Barve S, Hill DB. Cytokines and alcoholic liver disease. Alcohol Health Res World. 1997;21(4):317-20.

Shayakhmetova GM, Bondarenko LB, Matvienko AV, Kovalenko VM. Chronic alcoholism-mediated metabolic disorders in albino rat testes. Interdiscip Toxicol. 2014 Sep;7(3):165-72. DOI: 10.2478/intox-2014-0023

Shayakhmetova GM, Bondarenko LB, Kovalenko VM, Kharchenko OI, Bohun LI, Omelchenko YO. Multiparameter rodent chronic model for complex evaluation of alcoholism-mediated metabolic violations. J Basic Clin Physiol Pharmacol. 2015 Jan;26(1):43-51. DOI: 10.1515/jbcpp-2013-0163

Shayakhmetova GM, Bondarenko LB, Matvienko AV, Kovalenko VM. Correlation between spermatogenesis disorders and rat testes CYP2E1 mRNA contents under experimental alcoholism or type I diabetes. Adv Med Sci. 2014 Sep;59(2):183-9. DOI: 10.1016/j.advms.2014.03.004

Shayakhmetova GM, Bondarenko LB, Kovalenko VM, Ruschak VV. CYP2E1 testis expression and alcohol-mediated changes of rat spermatogenesis indices and type I collagen. Arh Hig Rada Toksikol. 2013;64(2):51-60. DOI: 10.2478/10004-1254-64-2013-2313

Bondarenko LB, Shayakhmetova GM, Kovalenko VM, Kharchenko OI , Bohun LI, Omelchenko YO. Chronic alcoholism-mediated metabolic violations in albino rats brain. Int J Biochem Res Rev. 2014;4(3):269-83. DOI: 10.9734/IJBCRR/2014/8809

Lamas-Paz A, Hao F, Nelson LJ, Vázquez MT, Canals S, Gómez Del Moral M, et al. Alcoholic liver disease: Utility of animal models. World J Gastroenterol. 2018 Dec 7;24(45):5063-75. DOI: 10.3748/wjg.v24.i45.5063

Galkin A, Komar A, Gorshunov Y, Besarab A, Soloviova V. New monoclonal antibodies to the prostate-specific antigen: obtaining and studying biological properties. J Microbiol Biotechnol Food Sci. 2019;9(3):573-7. DOI: 10.15414/jmbfs.2019/20.9.3.573-577

Jäger W, Correia MA, Bornheim LM, Mahnke A, Hanstein WG, Xue L, et al. Ethynylestradiol-mediated induction of hepatic CYP3A9 in female rats: implication for cyclosporine metabolism. Drug Metab Dispos. 1999 Dec;27(12):1505-11.

Lankford SM, Bai SA, Goldstein JA. Cloning of canine cytochrome P450 2E1 cDNA: identification and characterization of two variant alleles. Drug Metab Dispos. 2000 Aug;28(8):981-6.

Costa LG, Hodgson E, Lawrence DA, Ozolins TR, Reed DJ, Greenlee WF, editors. Current Protocols in Toxicology. John Wiley & Sons Inc; 2005. 2759 p.

Bondarenko L, Karatzuba T, Shayakhmetova G, Voronina A, Matvienko A, Strelkov I, et al. Specificity of metabolic syndrome model reproduction at pubertal and adult male rats. Rom J Diabetes Nutr Metab Dis. 2015;22(3):251-60. DOI: 10.1515/rjdnmd-2015-0031

Bondarenko LB, Saprykina NA, Kovalenko VM. Lung and spleen contents of free amino acids after pyrazinamide treatment. Acta Toxicol. 2006;14(1-2):79-86.

Glantz SA Primer of biostatistics. 7th edition. New York: McGraw Hill/Medical; 2011. 320 p.

Wang X, Lu Y, Cederbaum AI. Induction of cytochrome P450 2E1 increases hepatotoxicity caused by Fas agonistic Jo2 antibody in mice. Hepatology. 2005 Aug;42(2):400-10. DOI: 10.1002/hep.20792

Brahadeeswaran S, Tamizhselvi R. Consequence of alcohol intoxication-mediated efferocytosis impairment. Front Immunol. 2024 Jul 22;15:1386658. DOI: 10.3389/fimmu.2024.1386658

Miñana JB, Gómez-Cambronero L, Lloret A, Pallardó FV, Del Olmo J, Escudero A, et al. Mitochondrial oxidative stress and CD95 ligand: a dual mechanism for hepatocyte apoptosis in chronic alcoholism. Hepatology. 2002 May;35(5):1205-14. DOI: 10.1053/jhep.2002.32969

Kollár R, Sturdík E, Sablatúrová E. Biochemical, morphological and cytochemical studies of enhanced autolysis of Saccharomyces cerevisiae. 1. Biochemical studies. Folia Microbiol (Praha). 1993;38(6):473-8. DOI: 10.1007/BF02814398

Palmer E, Tyacke R, Sastre M, Lingford-Hughes A, Nutt D, Ward RJ. Alcohol hangover: Underlying biochemical, inflammatory and neurochemical mechanisms. Alcohol Alcohol. 2019 May 1;54(3):196-203. DOI: 10.1093/alcalc/agz016

Bloom JC, Loehr AR, Schimenti JC, Weiss RS. Germline genome protection: implications for gamete quality and germ cell tumorigenesis. Andrology. 2019 Jul;7(4):516-26. DOI: 10.1111/andr.12651

Xu W, Guo G, Li J, Ding Z, Sheng J, Li J, et al. Activation of Bcl-2-caspase-9 apoptosis pathway in the testis of asthmatic mice. PLoS One. 2016 Mar 3;11(3):e0149353. DOI: 10.1371/journal.pone.0149353

Muthusami KR, Chinnaswamy P. Effect of chronic alcoholism on male fertility hormones and semen quality. Fertil Steril. 2005 Oct;84(4):919-24. DOI: 10.1016/j.fertnstert.2005.04.025

Aitken RJ, Baker MA. Causes and consequences of apoptosis in spermatozoa; contributions to infertility and impacts on development. Int J Dev Biol. 2013;57(2-4):265-72. DOI: 10.1387/ijdb.130146ja

Sinha Hikim AP, Swerdloff RS. Hormonal and genetic control of germ cell apoptosis in the testis. Rev Reprod. 1999 Jan;4(1):38-47. DOI: 10.1530/ror.0.0040038

Khan AJ, Sharma A, Choudhuri G, Parmar D. Induction of blood lymphocyte cytochrome P450 2E1 in early stage alcoholic liver cirrhosis. Alcohol. 2011 Feb;45(1):81-7. DOI: 10.1016/j.alcohol.2010.08.007

Kumar S, Earla R, Jin M, Mitra AK, Kumar A. Effect of ethanol on spectral binding, inhibition, and activity of CYP3A4 with an antiretroviral drug nelfinavir. Biochem Biophys Res Commun. 2010;402(1):163-7. DOI: 10.1016/j.bbrc.2010.10.014

Roberts BJ, Shoaf SE, Song BJ. Rapid changes in cytochrome P4502E1 (CYP2E1) activity and other P450 isozymes following ethanol withdrawal in rats. Biochem Pharmacol. 1995 May 26;49(11):1665-73. DOI: 10.1016/0006-2952(95)00098-k

Rowlands JC, Wang H, Hakkak R, Ronis MJ, Strobel HW, Badger TM. Chronic intragastric infusion of ethanol-containing diets induces CYP3A9 while decreasing CYP3A2 in male rats. J Pharmacol Exp Ther. 2000 Nov;295(2):747-52.

Kim YJ, Park JE, Chung JY, Kim JY, Lee SG, Lee SJ, et al. Constitutive expression of cytochrome P450 1B1 endows testicular Leydig cells with susceptibility to 7,12-dimethylbenzanthracene-induced cell death. J Toxicol Sci. 2022;47(8):317-26. DOI: 10.2131/jts.47.317

Zhou SF. Drugs behave as substrates, inhibitors and inducers of human cytochrome P450 3A4. Curr Drug Metab. 2008 May;9(4):310-22. DOI: 10.2174/138920008784220664

Fau D. Imbalance through lysine excess and correction by a threonine supplement, as a function of nutritional status. Ann Nutr Aliment. 1975;29(4):321-35.

Li JY. Sequential changes in free amino acid pool in burned rabbits. Zhonghua Zheng Xing Shao Shang Wai Ke Za Zhi. 1991 Sep;7(3):208-11.

Murakami H, Shimbo K, Takino Y, Kobayashi H. Combination of BCAAs and glutamine enhances dermal collagen protein synthesis in protein-malnourished rats. Amino Acids. 2013 Mar;44(3):969-76. DOI: 10.1007/s00726-012-1426-4

Reilly ME, Mantle D, Richardson PJ, Salisbury J, Jones J, Peters TJ, et al. Studies on the time-course of ethanol's acute effects on skeletal muscle protein synthesis: comparison with acute changes in proteolytic activity. Alcohol Clin Exp Res. 1997 Aug;21(5):792-8.

Gardlik R, Palffy R, Hodosy J, Kopani M, Brezova V, Celec P. Similar effects of chronic voluntary alcohol intake and high dose superoxide dismutase gene delivery on oxidative and carbonyl stress in rats. Gene Ther Mol Biol. 2012;14:49-55.

Marks DB. Biochemistry. Baltimore: Williams & Wilkins; 1994. p. 234-49.

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Published

2024-12-09

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Bondarenko L, Shayakhmetova G, Kharchenko O, Kalachinskaya M, Serhiichuk N, Kovalenko V, Besarab O. Effects of Acute Alcohol Intoxication on Testicular Dna Stability, Gene Expression of Cytochromes CYP3A and CYP2E1, and Serum Pool of Free Amino Acids in Rats. Innov Biosyst Bioeng [Internet]. 2024Dec.9 [cited 2024Dec.21];8(4):3-12. Available from: https://ibb.kpi.ua/article/view/317136

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