Biochemical markers of the functional state of liver in rats fed diets with different protein and sucrose content
The study of mechanisms of the metabolic disorders in conditions of deficiency or excess of individual nutrients in the diet is a live issue. The influence of the simultaneous excess sucrose intake and protein deficiency in the diet on the functional state of the liver remains poorly understood.
The aim of the research was to study the rate of generation of the superoxide radicals, the content of triglycerides and glycogen in the liver, as well as the activity of enzymatic markers of the liver state in rats fed diets with different protein and sucrose content.
Material and methods. The studies were conducted over 28 days on 48 white non-linear rats, randomized into 4 groups: 1 - animals fed full-value semi-synthetic ration (14% protein); 2 - animals receiving low-protein ration (4.7% protein); 3 - animals receiving high-sucrose diet (40% sucrose), 4 - animals receiving low-protein high-sucrose diet. Serum sorbitol dehydrogenase activity was determined by the kinetic method in the reaction of NADH-dependent reduction of D-fructose to D-sorbitol. Serum alanine aminotransferase activity and aspartate aminotransferase was evaluated using a kit of reagents (Ukraine).
Results and discussion. It was found that in rats fed low protein diet, no changes in the de Ritis coefficient were observed, while the activity of sorbitol dehydrogenase in blood serum increased 1.7 fold. However, no changes in the rate of superoxide radical formation, as well as glycogen and triglyceride level in the liver were observed. In animals fed high-sugar diet, a rise in the de Ritis coefficient on the background of increased serum sorbitol dehydrogenase activity (more than 3.5 times) was revealed. At the same time, the rate of the superoxide radical formation in the liver mitochondria enhanced by 3 fold, with an increased accumulation of glycogen and triglycerides. The most pronounced changes in liver state were observed in animals fed low-protein/high-sugar diet: a marked increase in the de Ritis coefficient with a 5-fold increase in the activity of sorbitol dehydrogenase, and a 6-fold elevation in the intensity of the superoxide radical generation in liver mitochondria. The triglyceride content in the liver doubled, while the glycogen content remained at the level of control values.
Conclusion. The data obtained represent disturbances of the functional liver state as a consequence of the relatively short-term excessive consumption of sucrose, especially in combination with a alimentary protein deficiency. It was found that the leading factor in the formation of destructive changes in the liver was excessive sucrose consumption, while the concomitant protein deficiency exacerbated the functional changes in hepatocytes.
Keywords:protein deficiency, high sucrose diet, liver, de Ritis ratio, superoxide, triac-ylglycerol
For citation: Voloshchuk ON., Kopylchuk G.P., Holinei TYu. Biochemical markers of the functional state of liver in rats fed diets with different protein and sucrose content. Voprosy pitaniia [Problems of Nutrition]. 2019; 88 (6): 61-7. doi: 10.24411/0042-8833-2019-10065 (in Russian)
1. Wu G. Dietary protein intake and human health. Food Funct. 2016; 7 (3): 1251-65. doi: 10.1039/c5fo01530h
2. Tikole R.V., Kulkarni R., Uppinakudru S., et al. Nutritional deficiency disorders in paediatrics: an ayurvedic perspective. Int J Res Ayurveda Pharm. 2013; 4 (4): 605-7. doi: 10.7897/2277-4343.04431
3. Malta А., de Oliveira J.C., Ribeiro T.A., Tofolo L.P., et al. Low-protein diet in adult male rats has long-term effects on metabolism. J Endocrinol. 2014; 221 (2): 285-95. doi: 10.1530/JOE-13-0473
4. Kitada M., Ogura Y., Monno I., Koya D. The impact of dietary protein intake on longevity and metabolic health. EBioMedicine. 2019; 43: 632-40. URL: https://doi.org/10.1016/j.ebiom.2019.04.005
5. Maciejczyk M., Matczuk J., Zendzian-Piotrowska M., et al. Eight-week consumption of high-sucrose diet has a pro-oxidant effect and alters the function of the salivary glands of rats. Nutrients. 2018; 10 (10); 1-19. doi: 10.3390/nu10101530
6. Ragab S.M., Abd Elghaffar S.Kh., El-Metwally T.H., et al. Effect of a high fat, high sucrose diet on the promotion of non-alcoholic fatty liver disease in male rats: the ameliorative role of three natural compounds. Lipids Health Dis. 2015; 14: 83. doi: 10.1186/s12944-015-0087-1
7. Chung M., Ma J., Patel K., et al. Fructose, high-fructose corn syrup, sucrose, and nonalcoholic fatty liver disease or indexes of liver health: a systematic review and meta-analysis. Am J Clin Nutr. 2014; 100 (3): 833-49. URL; https://doi.org/10.3945/ajcn.114.086314
8. Morsy M.D., Abdel-Razek H.A., Eid R.A., El-Naby W.M.H. Impact of different doses of sucrose on the liver function and ultrastructure in rats. Med J Cairo Univ. 2014; 82 (1): 133-44.
9. Voloshchuk O.N., Kopylchuk G.P., Kadayskaia T.G. State of the energy-supply system of the liver mitochondria under the conditions of alimentary deficiency of protein. Voprosy pitaniia [Problems of Nutrition]. 2014; 83 (3): 12-6. (in Russian)
10. Reeves P., Nielsen F., Fahey G. AIN-93 purified diets for laboratory rodents: Final Report of the American Institute of Nutrition Ad Hoc Writing Committee on the Reformulation of the AIN-76A Rodent Diet. J Nutr. 1993; 123 (11): 1939-51.
11. Fernandes-Lima F., Monte L., Nascimento F., Gregorio B. Short exposure to a high-sucrose diet and the first "hit" of nonalcoholic fatty liver disease in mice. Cells Tissues Organs. 2016; 201 (6): 46472. doi: 10.1159/000446514
12. Rose C.I., Henderson A.R. Reaction rate assay of serum sorbitol dehydrogenase activity at 37 °C. Clin Chem. 1975; 21: 1619-24.
13. Kopylchuk G.P., Voloshchuk O.M. Peculiarities of the free radical processes in rat liver mitochondria under toxic hepatitis on the background of alimentary protein deficiency. Ukr Biochem J. 2016; 88 (2): 66-72.
14. Van Der Vies J. Two methods for the determination of glycogen in liver. Biochem J. 1954; 57 (3): 410-6. doi: 10.1042/bj0570410
15. Botros M., Sikaris K.A. The de ritis ratio: the test of time. Clin Biochem Rev. 2013; 34 (3): 117-30.
16. Ozer J., Ratner M., Shaw M., et al. The current state of serum biomarkers of hepatotoxicity. Toxicology. 2008; 245 (3): 194-205. doi: 10.1016/j.tox.2007.11.021
17. Singh A., Bhat T.K., Sharma O.P. Clinical biochemistry of hepa-totoxicity. J Clin Toxicol. 2011; S4: 1-19. doi: 10.4172/2161-0495. S4-001
18. Voloshchuk O.N., Kopylchuk G.P., Buchkovskaia I.M. Activity of the marker liver enzymes under the conditions of toxic hepatitis and alimentary deprivation of protein. Eksperimental’naya i klin-icheskaya gastoenterologiya [Experimental and Clinical Gastroenterology]. 2014; 108 (8): 96-100. (in Russian)
19. Lambis L.A., Solana T.J.B., Gastelbondo B.P., et al. Risk factors associated with nonalcoholic fatty liver disease in a Colombian Caribbean population. Rev Col Gastroenterol. 2016; 31 (2): 89-94.
20. Tan B.L., Norhaizan M.E., Liew W.P. Nutrients and oxidative stress: friend or foe? Oxid Med Cell Longev. 2018; 2018: 9719584. doi: 10.1155/2018/9719584