The effects of whey and pumpkin seed oil on blood biochemical parameters of liver function and lipid profile in rats chronically drinking low concentrations of ethanol
Keywords:whey protein, pumpkin seed oil, ethanol, liver biomarkers, rat
- Ethanol in alcoholic drinks causes liver injury. Cow whey and pumpkin seed oil have shown hepatoprotective effects in many studies.
- The effects of whey and pumpkin oil supplementation on the blood biochemical parameters of liver function in male Wistar rats subjected to chronic daily intake of 12% ethanol ad libitum were examined.
- Ethanol consummation in small doses for 6 weeks changed lipid metabolism, but did not induce extensive liver damage. Both whey and pumpkin seed oil showed some protective potential, but pumpkin seed oil and whey together with ethanol elevated lactate dehydrogenase.
Abstract: We studied the effects of whey and pumpkin seed oil supplementation on the biochemical parameters in blood serum of male rats after chronic ad libitum alcohol consumption. The levels of AST, ALT, total bilirubin, ALP, LDH, triglycerides, total cholesterol, HDL, LDL, VLDL, triglyceride/HDL ratio, total cholesterol/HDL ratio (cholesterol ratio) and LDL/HDL ratio (index of atherosclerosis) were determined in rats after six weeks of treatment with: (i) ethanol (12% ethanol, ad libitum), (ii) whey (2 g/kg per day), (iii) pumpkin seed oil (2 mL/kg per day), (iv) both ethanol and whey, and (v) both ethanol and pumpkin seed oil. The results showed no changes in the levels of AST, ALT, total bilirubin, ALP, total cholesterol, HDL and VLDL in alcoholic rats when compared to the controls (fed with a standard laboratory diet ad libitum) and rats supplemented with whey and pumpkin seed oil. Our results suggest that alcohol consumption in small doses for 6 weeks changes lipid metabolism and significantly elevates the LDL/HDL ratio (index of atherosclerosis) but does not induce extensive liver damage. Ethanol consumption in our experimental conditions lowered the triglyceride level as well as the triglyceride/HDL ratio, suggesting lipid redistribution and the induction of some cardio-protective effect. However, ethanol induced a higher index of atherosclerosis. Pumpkin seed oil showed some protective potential in alcoholic rats by lowering the total cholesterol/HDL ratio, but it elevated the LDH. Whey consumption prevented elevation of the atherosclerosis index, pointing to its protective role, probably through the redistribution of lipids. However, whey in combination with ethanol elevated LDH.
Received: March 20, 2018; Revised: March 30, 2018; Accepted: March 30, 2018; Published online: April 17, 2018
How to cite this article: Radić I, Nestorović V, Mijović M, Tatalović N, Joksimović B, Lukić V, Mitić M, Adžić M, Blagojević D, Veličković S, Bulajić S, Đerković B, Mirić M, Janićijević-Hudomal S. The effects of whey and pumpkin seed oil on blood biochemical parameters of liver function and lipid profile in rats chronically drinking low concentrations of ethanol. Arch Biol Sci. 2017;70(3):…
Pari L. KarthiKesan K. Protective role of caffeic acid against alcohol-induced biochemical changes in rats. Fundam Clin Pharmacol. 2007;21(4):355-61.
Dandona P, Aljada A, Bandyopadhyay A. Inflammation: the link between insulin resistance, obesity and diabetes. Trends Immunol. 2004;25(1):4-7.
Thiele G, Freeman T, Klassen L. Immunologic Mechanisms of Alcoholic Liver Injury. Semin Liver Dis. 2004;24(3):273-87.
Albano E. Alcohol, oxidative stress and free radical damage. Proc Nutr Soc. 2006;65(3):278-90.
Dey A, Cederbaum AI. Alcohol and oxidative liver injury. Hepatology. 2006;43(2):63-74.
Das KS, Vasudevan MD. Effect of ethanol on liver antioxidant defense systems: A dose dependent study. Indian J Clin Biochem. 2005;20(1):80-4.
Tang Y, Banan A, Forsyth BC, Fields ZJ, Lau KC, Zhang JL, Keshavarzian A. Effect of alcohol on miR-212 exspression in intestinal epithelial cells and its potential role in alcoholic liver disease. Alcohol Clin Exp Res. 2008;32(2):355-64.
Gyamfi AM, Wan YY. Pathogenesis of alcoholic liver disease: the role of nuclear receptors. Exp Biol Med. 2010;235(5):547-60.
Klop B, do Rego AT, Cabezas MC. Alcohol and plasma triglycerides. Curr Opin Lipidol. 2013;24(4):321-6.
Zhang JW, Tong X, Wan Z, Wang Y, Qin LQ, Szeto IM. Effect of whey protein on blood lipid profiles: a meta-analysis of randomized controlled trials. Eur J Clin Nutr. 2016;70(8):879-85.
Gad AS, Khadrawy YA, El-Nekeety AA, Mohamed SR, Hassan NS, Abdel-Wahhab MA. Antioxidant activity and hepatoprotective effects of whey protein and Spirulina in rats. Nutrition. 2011;27(5):582-9.
Ilyas UK, Katare DP, Aeri V. Comparative evaluation of standardized alcoholic, hydroalcoholic, and aqueous extracts of Phyllanthus maderaspatensis Linn. against galactosamine-induced hepatopathy in albino rats. Pharmacogn Mag. 2015;11(42):277-82.
Eraslan G, Kanbur M, Aslan Ö, Karabacak M. The antioxidant effects of pumpkin seed oil on subacute aflatoxin poisoning in mice. Environ Toxicol. 2013;28(12):681-8.
Vetriselvan S, Rajamanikkam V, Devi Subasini P, Arun G. Comparative evaluation of hepatoprotective activity of andrographis paniculata and silymarin in ethanol induced hepatotoxicity in albino wistar rats. Pharm Lett. 2010;2(6):52-9.
Zhao Z, Pan D, Wu Z, Sun Y, Guo Y, Zeng X. Antialcoholic liver activity of whey fermented by Lactobacillus casei isolated from koumiss. J Dairy Sci. 2014;97(7):4062-71.
Abou Seif H. Ameliorative effect of pumpkin oil (Cucurbita pepo L.) against alcohol-induced hepatotoxicity and oxidative stress in albino rats. Beni-Suef Univ J Basic Appl. 2014;3(3):178-85.
Kaya S, Karakus A, Mergen G, Dural E, Aliyev V, Yalçin S, Söylemezoglu T, Kayaalti Z. Simultaneous Headspace-GC–FID Analysis for Methanol and Ethanol in Blood, Saliva, and Urine: Validation of Method and Comparison of Specimens. LC GC. 2010;28(7):540-3.
Hinkle ED, Wiersma W, Jurs GS. Applied Statistics for Behavioral Sciences, 5th ed. Boston, MA, USA: Houghton Mifflin Company; 2002.
Whitfield JB, Zhu G, Heath AC, Powell LW, Martin NG. Effects of alcohol consumption on indices of iron stores and of iron stores on alcohol intake markers. Alcohol Clin Exp Res. 2001;25(7):1037-45.
Alatalo PI, Koivisto HM, Hietala JP, Puukka KS, Bloigu R, Niemela OJ. Effect of moderate alcohol cosumption on liver enzymes increases with increasing body mass index. Am J Clin Nutr. 2008;88(4):1097-103.
Liangpunsakul S, Qi R, Crabb DW, Witzmann F. Relationship Between Alcohol Drinking and Aspartate Aminotransferase: Alanine Aminotransferase (AST:ALT) Ratio, Mean Corpuscular Volume (MCV), Gamma-Glutamyl Transpeptidase (GGT), and Apolipoprotein A1 and B in the U.S. Population. J Stud Alcohol Drugs. 2010;71(2):249-52.
Li YM, Chen SH, Yu CH, Zhang Y, Xu GY. Effect of acute alcoholism on hepatic enzymes and oxidation/antioxidation in rats. Hepatobiliary Pancreat Dis Int. 2004;3(2):241-4.
Hakucho A, Liu J, Horiguchi N, Liu X, Fujimiya T. The effects of chronic alcohol exposure on progression of liver injury in spontaneously hypertensive rats; preliminary study. Nihon Arukoru Yakubutsu Igakkai Zasshi. 2013;48(3):216-22.
Bourogaa E, Nciri R, Mezghani-Jarraya R, Racaud-Sultan C, Damak M, El Feki A. Antioxidant activity and hepatoprotective potential of Hammada scoparia against ethanol-induced liver injury in rats. J Physiol Biochem. 2013;69(2):227-37.
Kasdallah-Grissa A, Nakbi A, Koubaa N, El-Fazaâ S, Gharbi N, Kamoun A, Hammami, M. Dietary virgin olive oil protects against lipid peroxidation and improves antioxidant status in the liver of rats chronically exposed to ethanol. Nutr Res. 2008;28(7):472-9.
Sato C, Hasumura Y, Takeuchi J. Interaction of Ethanol with Drugs and Xenobiotics. In: Seitz HK, Kommerell B, editors. Alcohol Related Diseases in Gastroenterology. Berlin, Heidelberg New York, Tokyo: Springer Verlag; 1985. p. 172-84.
Ideo G, De Franchis R, Del Ninno E, Cocucci C, Dioguardi N. Increase of some rat liver microsomal enzymes as a consequence of prolonged alcohol intake: comparison with the effect of phenobarbitone. Enzyme. 1971;12(4):473-80.
Yost GS, Finley BL. Ethanol as an inducer of UDP-glucuronyltransferase:a comparison with phenobarbital and 3-methylcholanthrene induction in rabbit hepatic microsomes. Biochem Biophys Res Commun. 1983;111(1):219-23.
Lieber CS. Ethanol metabolism and pathophysiology of alcoholic liver disease. In: Seitz HK, Kommerell B, editors. Alcohol related diseases in gastroenterology. Berlin, Heidelberg, New York, Tokyo: Springer Verlag; 1985. p. 10-47.
Yamada S, Mak KM, Lieber CS. Chronic Ethanol Consumption Alters Rat Liver Plasma Membranes and Potentiates Release of Alkaline Phosphatase. Gastroenterology. 1985;88(6):1799-806.
Evans WH. Preparation of plasma membranes from certain tissues and cell lines. In: Evans WH, editor. Preparation and characterisation of mammalian plasma membranes. Part I. Amsterdam: Elsevier/North Holland Biomedical; 1978. p. 177-83.
Horner F, Kellen JA, Kingston E, Maharaj N, Malkin A. Dynamic changes of serum gamma-glutamyl transferase in chronic alcoholism. Enzyme. 1979;24(4):217-23.
Nishimura M, Teschke R. Effect of chronic alcohol consumption on the activities of liver plasma membrane enzymes: gamma-glutamyltransferase, alkaline phosphatase and 5’-nucleotidase. Biochem Pharmacol. 1982;31(3):377-81.
Markert CL. Izozymes II physiological funtions. New York: Academic press INC; 1975.
Chen WC, Huang WC, Chiu CC, Chang YK, Huang CC. Whey Protein Improves Exercise Performance and Biochemical Profiles in Trained Mice. Med Sci Sports Exerc. 2014;46(8):1517-24.
Brien S, Ronksley P, Turner B, Mukamal K, Ghali W. Effect of alcohol consumption on biological markers associated with risk of coronary heart disease: systematic review and meta-analysis of interventional studies. Br Med J. 2011;342:d636.
Tolstrup JS, Gronbaek M, Nordestgaard BG. Alcohol intake, myocardial infarction, biochemical risk factors, and alcohol dehydrogenase genotypes. Circ Cardiovasc Genet. 2009;2(5):507-14.
Kuusisto S, Peltola T, Laitinen M, Kumpula L, Mäkinen V, Salonurmi T, Hedberg P, Jauhiainen M, Savolainen M, Hannuksela M, Ala-Korpela M. The interplay between lipoprotein phenotypes, adiponectin, and alcohol consumption. Ann Med. 2011;44(5):513-22.
Ranković S, Popović T, Martačić J, Petrović S, Tomić M, Ignjatović Đ, Tovilović-Kovačević G, Glibetić M. Liver phospholipids fatty acids composition in response to different types of diets in rats of both sexes. Lipids Health Dis. 2017;16:94.
Popović T, Borozan S, Takić M, Kojadinović M, Rankovic S, Ranić M, de Luka S. Fatty Acid Composition and Oxidative Stress Parameters in Plasma after Fish Oil Supplementation in Aging. Croat Chem Acta. 2014;87(3):207-12.
Popović T, Borozan S, Arsić A, Martačić JD, Vučić V, Trbović A, Mandić L, Glibetić M. Fish oil supplementation improved liver phospholipids fatty acid composition and parameters of oxidative stress in male Wistar rats. J Anim Physiol Anim Nutr. 2011;96(6):1020-9.
Gupta S, Figueredo VM. Alcohol and lipids. OA Alcohol. 2014;2(1):3.
De Oliveira E, Silva ER, Foster D, McGee Harper M, Seidman CE, Smith JD, Breslow JL, Brinton EA. Alcohol consumption raises HDL cholesterol levels by increasing the transport rate of apolipoproteins A-I and A-II. Circulation. 2000;102(19):2347-52.
Sabesin SM, Hawkins HL, Kuiken L, Ragland JB. Abnormal plasma lipoproteins and lecithin-cholesterol acyltransferase deficiency in alcoholic liver disease. Gastroenterology. 1977;72:510-8.
Serdyuk AP, Metelskaya VA, Ozerova IN, Kovaltchouk NV, Olferiev AM, Bubnova MG, Perova NV, Jauhiainen M, Lasselin C, Castro G. Effects of alcohol on the major steps of reverse cholesterol transport. Biochemistry (Mosc). 2000;65:1310-5.
Hannuksela M, Marcel YL, Kesaniemi YA, Savolainen MJ. Reduction in the concentration and activity of plasma cholesteryl ester transfer protein by alcohol. J Lipid Res. 1992;33:737-44.
Hendriks HF, Veenstra J, van Tol A, Groener JE, Schaafsma G. Moderate doses of alcoholic beverages with dinner and postprandial high density lipoprotein composition. Alcohol and alcoholism. 1998;33(4):403-10.
Riemens SC, van Tol A, Hoogenberg K, van Gent T, Scheek LM, Sluiter WJ, Dullaart RP. Higher high density lipoprotein cholesterol associated with moderate alcohol consumption is not related to altered plasma lecithin:cholesterol acyltransferase and lipid transfer protein activity levels. Clin Chim Acta. 1997;258(1):105-15.
Ordovas JM, Cupples LA, Corella D, Otvos JD, Osgood D, Martinez A, Lahoz C, Coltell O, Wilson PW, Schaefer EJ. Association of cholesteryl ester transferprotein-TaqIB polymorphism with variations in lipoprotein subclasses and coronary heart disease risk: the Framingham study. Arterioscler Thromb Vasc Biol. 2000;20(5):1323-9.
Kenneth JM, Rachel HM, Lewis HK, Russell PT, Richard AK, Murray AM, David SS. Alcohol Consumption and Lipoprotein Subclasses in Older Adults. The Journal of Clinical Endocrinology & Metabolism. 2007;92(7):2559-66.
Otvos JD, Collins D, Freedman DS, Shalaurova I, Schaefer EJ, McNamara JR, Bloomfield HE, Robins SJ. Low-density lipoprotein and high-density lipoprotein particle subclasses predict coronary events and are favorably changed by gemfibrozil therapy in the Veterans Affairs High-Density Lipoprotein Intervention Trial. Circulation. 2006;113(12):1556-63.
Freedman DS, Otvos JD, Jeyarajah EJ, Barboriak JJ, Anderson AJ, Walker JA. Relation of lipoprotein subclasses as measured by proton nuclear magnetic resonance spectroscopy to coronary artery disease. Arterioscler Thromb Vasc Biol. 1998;18(7):1046-53.
Blum C, Levy R, Eisenberg S, Hall M, Goebel R, Berman M. High Density Lipoprotein Metabolism in Man. J Clin Invest. 1977;60(4):795-807.
Grundy SM. Effects of polyunsaturated fats on lipid metabolism in patients with hypertriglyceridemia. J Clin Invest. 1975;55(2):269-82.
Nishiwaki M, Ishikawa T, Ito T, Shige H, Tomiyasu K, Nakajima K, Kondo K, Hashimoto H, Saitoh K, Manabe M, Miyajima E, Nakamura H. Effects of alcohol on lipoprotein lipase, hepatic lipase, cholesteryl ester transfer protein, and lecithin:cholesterol acyltransferase in high-density lipoprotein cholesterol elevation. Atherosclerosis. 1994;111(1):99-109.
Baraona E, Lieber SC. Effects of chronic ethanol feeding on serum lipoprotein metabolism in the rat. J Clin Invest. 1970;49(4):769-78.
Iseri OA, Lieber SC, Gottlieb SL. The ultrastructure of fatty liver induced by prolonged ethanol ingestion. Am J Pathol. 1966;48:535-55.
Lane BP, Lieber CS. Ultrastructural alterations in human hepatocytes following ingestion of ethanol with adequate diets. Am J Pathol. 1966;49(4):593-603.
Nanji AA, Mendenhall CL, French SW. Beef fat prevents alcoholic liver disease in the rat. Alcohol Clin Exp Res. 1989;13(1):15-9.
Nanji AA, French SW. Dietary linoleic acid is required for development of experimentally induced alcoholic liver injury. Life Sci. 1989;44(3):223-7.