Acute Alcohol Tissue Damage: Protective Properties of Betaine

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Lucy Petagine
Hannah Everitt
Victor Preedy
Roy Sherwood
Vinood Patel


adolescents; antioxidants; binge drinking; liver; oxidative stress


Teenage binge drinking is a major health issue; however, there is a paucity of data on liver injury. Herein, we investigated how acute ethanol affects juvenile hepatic cells through changes in oxidative stress, apoptosis, and liver function, as well as the ability of betaine, which can replen-ish the antioxidant glutathione and mitigate oxidative injury. Juvenile male Wistar rats were given either water or betaine (2% w/v) for 6 days and treated with either saline 0.15 mol/L NaCl or ethanol (75 mmol/kg bodyweight). After 24 h, liver enzymes, oxidative damage, apoptosis, and parameters of antioxidant enzyme activity were examined. Acute ethanol increased hepatic enzymes (99%, P < 0.05). Total protein and albumin levels were reduced by 14 and 18% (P < 0.001), respectively, which was prevented by betaine treatment. Cytosolic cytochrome c increased by 59% (P < 0.05), corresponding to a decrease in mitochondrial cytochrome c content, which was ameliorated with betaine. Cytosolic glutathione peroxidase was reduced with alcohol (P < 0.05) and was prevented with betaine. Subtle changes were observed in catalase, superoxide dismutase, glutathione reductase, and complex I activity after ethanol treatment. In summary, whilst juvenile animals appear to have higher basal levels of antioxidant enzymes, betaine conferred some protection against alcohol-induced oxidative stress.


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1. Mathurin P, Hadengue A, Bataller R, Addolorato G, Burra P, Burt A, et al. EASL clinical practical guidelines: Management of alcoholic liver disease. J Hepatol. 2012;57:399–420. http://dx.
2. Chung T, Creswell KG, Bachrach R, Clark DB, Martin CS. Adolescent binge drinking. Alcohol Res Curr Rev. 2018;39:5–15.
3. Hibell B, Stergar E, Dernovšc?ek Hafner N. The 2011 ESPAD report: Substance use among students in 36 European coun-tries. The Swedish Council for Information on Alcohol and other Drugs. 2012.
4. Hibell B, Andersson B, Bjarnason T, Ahlström S, Balakireva O,
Kokkevi A, et al. The ESPAD report 2003: Alcohol and other drug use among students in 35 European countries. The Swedish Council for Information on Alcohol and other Drugs. 2004.
5. Clark Goings T, Salas-Wright CP, Belgrave FZ, Nelson EJ, Harezlak J, Vaughn MG. Trends in binge drinking and alcohol abstention among adolescents in the US, 2002–2016. Drug Alcohol Depend. 2019;200:115–123. http://dx.doi. org/10.1016/j.drugalcdep.2019.02.034
6. Tapia-Rojas C, Torres AK, Quintanilla RA. Adolescence binge alcohol consumption induces hippocampal mitochondrial impairment that persists during the adulthood. Neuroscience. 2019;406:356–68. neuroscience.2019.03.018
7. Bailey SM, Cunningham CC. Acute and chronic ethanol increases reactive oxygen species generation and decreases viabil-ity in fresh, isolated rat hepatocytes. Hepatology. 1998;28:1318– 26.
8. Bailey SM, Pietsch EC, Cunningham CC. Ethanol stimulates the production of reactive oxygen species at mitochondrial com-plexes I and III. Free Radic Biol Med. 1999;27:891–900. http://
9. Ismail NA, Okasha SH, Dhawan A, Abdel-Rahman AO, Shaker OG, Sadik NA. Antioxidant enzyme activities in hepatic tissue from children with chronic cholestatic liver disease. Saudi J Gastroenterol. 2010;16:90–4. http://dx.doi. org/10.4103/1319-3767.61234
10. Chrobot AM, Szaflarska-Szczepanik A, Drewa G. Antioxidant defense in children with chronic viral hepatitis B and C. Med Sci Monit Int Med J Exp Clin Res. 2000;6:713–18.
11. Moreno I, Pichardo S, Jos A, Gómez-Amores L, Mate A, Vazquez CM, et al. Antioxidant enzyme activity and lipid peroxidation in liver and kidney of rats exposed to microcystin-LR administered intraperitoneally. Toxicon. 2005;45:395–402.
12. Kurutas EB. The importance of antioxidants which play the role in cellular response against oxidative/nitrosative stress: Current state. Nutr J. 2016;15:71. s12937-016-0186-5
13. Purohit V, Abdelmalek MF, Barve S, Benevenga NJ, Halsted CH, Kaplowitz N, et al. Role of S-adenosylmethionine, folate, and betaine in the treatment of alcoholic liver disease: Summary of a symposium. Am J Clin Nutr. 2007;86:14–24.
14. Barak AJ, Beckenhauer HC, Tuma DJ. Betaine effects on hepatic methionine metabolism elicited by short-term ethanol feeding. Alcohol. 1996;13:483–486. http://dx.doi. org/10.1016/0741-8329(96)00040-7
15. Kharbanda KK, Todero SL, King AL, Osna NA, McVicker BL, Tuma DJ, et al. Betaine treatment attenuates chronic ethanolin-duced hepatic steatosis and alterations to the mitochondrial respiratory chain proteome. Int J Hepatol. 2012;2012:962183.
16. Barak AJ, Beckenhauer HC, Mailliard ME, Kharbanda KK, Tuma DJ. Betaine lowers elevated S-adenosylhomocysteine levels in hepatocytes from ethanol-fed rats. J Nutr. 2003;133:2845– 8.
17. Kanbak G, I?nal M, Bayçu C. Ethanol-induced hepatotoxicity and protective effect of betaine. Cell Biochem Funct. 2001;19:281–5.
18. Kharbanda KK, Mailliard ME, Baldwin CR, Beckenhauer HC, Sorrell MF, Tuma DJ. Betaine attenuates alcoholic steatosis by restoring phosphatidylcholine generation via the phos-phatidylethanolamine methyltransferase pathway. J Hepatol. 2007;46:314–21.
19. Kim SJ, Jung YS, Kwon DY, Kim YC. Alleviation of acute ethanol-induced liver injury and impaired metabolomics of S-containing substances by betaine supplementation. Biochem Biophys Res Commun. 2008;368:893–8. http://dx.doi. org/10.1016/j.bbrc.2008.02.003
20. Balkan J, Öztezcan S, Küçük M, Çevikbas¸ U, Koçak-Toker N, Uysal M. The effect of betaine treatment on triglyceride levels and oxidative stress in the liver of ethanol-treated guinea pigs. Exp Toxicol Pathol. 2004;55:505–9. http://dx.doi. org/10.1078/0940-2993-00347
21. Kharbanda KK, Rogers II DD, Mailliard ME, Siford GL, Barak AJ, Beckenhauer HC, et al. A comparison of the effects of betaine and S-adenosylmethionine on ethanol-induced changes in methionine metabolism and steatosis in rat hepatocytes. J Nutr. 2005;135:519–24.
22. Kharbanda KK, Todero SL, Ward BW, Cannella JJ, Tuma DJ. Betaine administration corrects ethanol-induced defective VLDL secretion. Mol Cell Biochem. 2009;327:75–8. http://dx.
23. Alirezaei M, Jelodar G, Niknam P, Ghayemi Z, Nazifi S. Betaine prevents ethanol-induced oxidative stress and reduces total homocysteine in the rat cerebellum. J Physiol Biochem. 2011;67:605–12.
24. Patel VB, Spencer CH, Young TA, Lively MO, Cunningham  CC. Effects of 4-hydroxynonenal on mitochon-drial 3-hydroxy-3-methylglutaryl (HMG-CoA) synthase. Free Radic Biol Med. 2007;43:1499–507. freeradbiomed.2007.08.004
25. Preedy VR, Gove CD, Panos MZ, Sherwood R, Portmann B, Williams R, et al. Liver histology, blood biochemistry and RNA, DNA and subcellular protein composition of various skeletal muscles of rats with experimental cirrhosis: Implications for alcoholic muscle disease. Alcohol Alcohol. 1990;25:641–9.
26. Tietze F. Enzymic method for quantitative determination of nano-gram amounts of total and oxidized glutathione: Applications to mammalian blood and other tissues. Anal Biochem. 1969;27:502– 22.
27. Bar-Or D, Rael LT, Lau EP, Rao NKR, Thomas GW, Winkler JV, et al. An analog of the human albumin N-terminus (Asp-Ala-His-Lys) prevents formation of copper-induced reactive oxygen species. Biochem Biophys Res Commun. 2001;284:856– 62.
28. Yemis¸ci M, Gürsoy-Özdemir Y, Caban S, Bodur E, Çapan Y, Dalkara T. Transport of a caspase inhibitor across the blood-brain barrier by chitosan nanoparticles. In: Methods in enzymology. Editor, Nejat Düzgünes¸ Academic Press; Netherlands:Elsevier. 2012. p. 253–69.
29. Kim SK, Kim YC. Attenuation of bacterial lipopolysaccharide-induced hepatotoxicity by betaine or taurine in rats. Food Chem Toxicol. 2002;40:545–9. S0278-6915(01)00102-8
30. Erman F, Balkan J, Çevikbas¸ U, Koçak-Toker N, Uysal M. Betaine or taurine administration prevents fibrosis and lipid peroxidation induced by rat liver by ethanol plus carbon tetrachloride intoxication. Amino Acids. 2004;27:199–205. http://dx.doi. org/10.1007/s00726-004-0105-5
31. Lang CH, Liu X, Nystrom G, Wu D, Cooney RN, Frost RA. Acute effects of growth hormone in alcohol-fed rats. Alcohol Alcohol. 2000;35:148–58. alcalc/35.2.148
32. Hayes KC, Pronczuk A, Cook MW, Robbins MC. Betaine in sub-acute and sub-chronic rat studies. Food Chem Toxicol. 2003;41:1685–700.
33. del Pilar Cabrales-Romero M, Márquez-Rosado L, Fattel-Fazenda S, Trejo-Solís C, Arce-Popoca E, Alemán-Lazarini L, et al. S-adenosyl-methionine decreases ethanol-induced apoptosis in primary hepatocyte cultures by a c-Jun N-terminal kinase activity-independent mechanism. World J Gastroenterol. 2006;12:1895–904.
34. Ferrari D, Stepczynska A, Los M, Wesselborg S, Schulze-Osthoff K. Differential regulation and ATP requirement for caspase-8 and caspase-3 activation during CD95- and anticancer drug-induced apoptosis. J Exp Med. 1998;188:979–84. http://
35. Young TA, Bailey SM, Van Horn C, Cunningham CC. Chronic ethanol consumption decreases mitochondrial and glycolytic production of ATP in liver. Alcohol Alcohol. 2006;41:254–60.
36. Nassir F, Ibdah JA. Role of mitochondria in alcoholic liver disease. World J Gastroenterol. 2014;20:2136–42. http://dx.doi. org/10.3748/wjg.v20.i9.2136
37. Andreoli SP. Mechanisms of endothelial cell ATP depletion after oxidant injury. Pediatr Res. 1989;25:97–101. http://dx.doi. org/10.1203/00006450-198901000-00021
38. Ghanbari-Niaki A, Désy F, Lavoie JM. Effects of phosphate injection on metabolic and hormonal responses to exercise in fructose-injected rats. Physiol Behav. 1999;67:747–52. http://dx.
39. Crompton M. The mitochondrial permeabitity transition pore and its role in cell death. Biochem J. 1999;341:233–49. http://dx.
40. Rasola A, Bernardi P. The mitochondrial permeability transition pore and its involvement in cell death and in disease patho-genesis. Apoptosis. 2007;12:815–33. s10495-007-0723-y
41. Cederbaum AI. Microsomal generation of reactive oxygen spe-cies and their possible role in alcohol hepatotoxicity. Alcohol Alcohol Suppl. 1991;1:291–6.
42. Vogt BL, Richie JP. Glutathione depletion and recovery after acute ethanol administration in the aging mouse. Biochem Pharmacol. 2007;73:1613–21. 01.033
43. Kade S, Herzog N, Schmidtke K-U, Küpper J-H. Chronic etha-nol treatment depletes glutathione regeneration capacity in hep-atoma cell line HepG2. J Cell Biotechnol. 2016;1:183–90. http://
44. Guerri C, Grisolía S. Changes in glutathione in acute and chronic alcohol intoxication. Pharmacol Biochem Behav. 1980;13:53–61.
45. Hoek JB, Cahill A, Pastorino JG. Alcohol and mitochondria: A dysfunctional relationship. Gastroenterology. 2002;122:2049–63.
46. Bailey SM, Patel VB, Young TA, Asayama K, Cunningham CC. Chronic ethanol consumption alters the glutathione/glutathione peroxidase-1 system and protein oxidation status in rat liver. Alcohol Clin Exp Res. 2001;25:726–33. http://dx.doi. org/10.1111/j.1530-0277.2001.tb02273.x
47. Sriram KI, Lakshmi CJ. Endurance exercise-induced alterations in antioxidant enzymes of old albino male rats. Curr Sci. 2001;80:921–3.
48. Mallikarjuna K, Shanmugam KR, Nishanth K, Wu MC, Hou  CW, Kuo CH, et al. Alcohol-induced deterioration in primary antioxidant and glutathione family enzymes reversed by exercise training in the liver of old rats. Alcohol. 2010;44:523–9.
49. Cahill A, Hershman S, Davies A, Sykora P. Ethanol feed-ing enhances age-related deterioration of the rat hepatic mitochondrion. Am J Physiol Liver Physiol. 2005;289:G1115– 23.
50. Meier P, Seitz HK. Age, alcohol metabolism and liver disease. Curr Opin Clin Nutr Metab Care. 2008;11:21–6. http://dx.doi. org/10.1097/MCO.0b013e3282f30564