Tag Archives: liver

Phenformin attenuates the oxidative-nitrosative stress in the liver of rats under long-term ethanol administration

A. Mykytenko1*, O. Akimov2, G. Yeroshenko3, K. Neporada1

1Department of Bioorganic and Biological Chemistry,
Poltava State Medical University, Poltava, Ukraine;
2Department of Pathophysiology, Poltava State Medical University, Poltava, Ukraine;
3Department of Medical Biology, Poltava State Medical University, Poltava, Ukraine;
*e-mail: mykytenkoandrej18@gmail.com

Received: 09 March 2024; Revised: 29 April 2024;
Accepted: 31 May 2024; Available on-line: 17 June 2024

Modulation of the AMP-activated protein kinase (AMPK) pathway activity is considered to be a promi­sing option in the development of approaches to chronic alcoholic hepatitis treatment. Phenformin, which is a biguanide, has been reported to increase AMPK activity. The aim of this work was to estimate the effect of phenformin as AMPK activator on the development of oxidative-nitrosative stress in the liver of rats under conditions of long-term ethanol administration. The experiments were performed on 24 male Wistar rats, divided into 4 groups: control; animals, which received phenformin hydrochloride orally at a dose of 10 mg/kg daily for 63 days; animals with a forced intermittent alcoholization for 5 days by intraperitoneal administration of 16.5% ethanol solution in 5% glucose at the rate of 4 ml/kg b.w. and subsequent transfer to 10% ethanol as the only source of drinking; animals with chronic alcohol hepatitis simulation and phenformin administration. Superoxide dismutase, catalase, NO synthase isoforms activity, superoxide anion radical production, concentration of malonic dialdehyde, peroxynitrite, nitrites, nitrosothiols concentration and oxidative modification of proteins (OMP) were estimated in liver homogenates. The increased production of oxygen and nitrogen active forms and OMP intensification in the liver of rats under long-term administration of ethanol was detected. Phenformin introduction under long-term ethanol administration was shown to limit the excess peroxynitrite formation and to prevent oxidative damage to rat liver proteins.

Free radical processes in the liver mitochondria of rats exposed to diethyl phthalate

O. V. Ketsa*, A. P. Husliakova, M. M. Marchenko

Yuriy Fedkovych Chernivtsi National University, Chernivtsi, Ukraine;
*e-mail: o.ketsa@chnu.edu.ua

Received: 11 October 2023; Revised: 28 November 2023;
Accepted: 01 February 2024; Available on-line: 26 February 2024

Diethyl phthalate (DEF) is a synthetic chemical widely used as plasticizer and additive in personal care and pharmaceutical products. Low-dose exposure to this xenobiotic over a long period contributes to its intake into the human body in a high doses. To date, studies of DEF influence on free radical processes in liver cells, in which it is not only metabolized but can also have a prooxidant effect, remain limited. The aim of our research was to determine ROS formation, the intensity of oxidative modification of proteins (OMP) and proteolytic activity in the mitochondrial fraction of liver tissue of rats exposed to diethyl phtalate. The experimental white outbred rats were divided into three groups: I – intact animals (control), II and III – rats administered DEF orally for three weeks at a doses that reflect the dose levels received by humans, namely 2.5 and 5.4 mg/kg b.w. respectively. The animals were euthanized on the 14th and 21st day after xenobiotic administration. The mitochondrial fraction from the rat liver was isolated and the level of superoxide and hydroxyl radicals, protein carbonyl derivatives, SH-groups and Schiff bases was determined. Proteolytic activity was assessed in the test of hemoglobin cleavage. It was shown that administration of DEF in a dose of 2.5 mg/kg initiated ROS generation and OMP intensification in the rat liver mitochondria only with prolonged administration for 21 days, whereas its administration in a dose of 5.4 mg/kg led to intensification of these processes already on day 14th followed by further amplification on day 21st. The activity of proteolytic enzymes in the mitochondrial fraction was found to be depended on the degree of DEF–induced OMP and was increased with minor OMP intensification but decreased with significant intensification of proteins free radical oxidation.

AUT-M enterosorbent stabilizes glutathione system in vincristine-treated rats with dimethylhydrazine-induced colon cancer

O. I. Kachur*, L. S. Fira, P. H. Lykhatskyі,
I. R. Bekus, M. V. Kyryliv

I. Horbachevsky Ternopil National Medical University, Ternopil, Ukraine;
*e-mail: oksana.kachur2012@gmail.com

Received: 15 August 2023; Revised: 17 October 2023;
Accepted: 01 December 2023; Available on-line: 18 December 2023

Colorectal cancer is one of the leading causes of mortality in the world. The search for new methods of therapy for this disease that could correct the state of oxidative stress during the development of neoplasms is up to date. The aim of this work was to study the level of reduced glutathione and the activity of glutathione-dependent enzymes in the development of 1,2 dimethylhydrazine-induced colon cancer in rats while treated with vincristine and the use of enterosorbent. To induce carcinogenesis, dimethylhydrazine was administered to male rats subcutaneously for 30 weeks at a dose of 7.2 mg/kg of body weight. The rats with induced colon cancer received entorosorbent per os at a dose of 0.2 g per 100 g of body weight daily for 21 days. After detoxification therapy, the rats were administered cytostatic vincristine daily at a dose of 0.23 mg/kg for 14 days. A decrease in the content of reduced glutathione, the activity of glutathione reductase and glutathione peroxidase in the blood and liver tissue of rats with colorectal cancer was established. The use of enterosorbent­ AUT-M was shown to be effective in stabilizing the indicators of the glutathione system in rats with induced colon cancer. Cytostatic vincristine did not significantly affect the change of the studied indicators, confirming the effectiveness of previous sorption measures.

Bioenergetic functions of mitochondria in liver, pancreatic acinar cells, and sperm cells of rats fed short-term high-fat or high-fat high-sugar diets

B. V. Manko1*, N. M. Kozopas1,2, H. M. Mazur1,
A. М. Voityk1, B. O. Manko1, V. V. Manko1

1Ivan Franko National University of Lviv,
Department of Human and Animal Physiology, Lviv, Ukraine;
2Danylo Halytsky Lviv National Medical University,
Department of Clinical Laboratory Diagnostics, Lviv, Ukraine;
*e-mail: bohdan.manko.ablb@lnu.edu.ua

Received: 26 September 2023; Revised: 23 October 2023;
Accepted: 27 October 2023; Available on-line: 06 November 2023

An unhealthy diet often is a cause of obesity, chronic inflammation, and metabolic disruption in multiple organs. However, the direct influence of elevated lipid or sugar consumption on liver, pancreatic, and sperm mitochondria is not well understood. The aim of the study was to investigate the functional activity of mitochondria of liver, pancreatic acinar cells, and sperm cells in rats on a short-term (7 weeks) diet with high fat or high fat and high sugar content. Male Wistar rats were on a basic, high-fat or high-fat high-sugar diet for 7 weeks. At the end of the experiment, visceral fat mass, blood glucose and lipids were measured. Mitochondrial functional activity was evaluated with oxygen consumption assay. In isolated pancreatic acinar cells, NAD(P)H autofluorescence and mitochondrial membrane potential were also studied. No difference in body mass was observed between the 3 groups at the end of the experiment. Visceral fat mass was slightly but significantly elevated in rats on a high-fat high-sugar diet. Both diets did not affect plasma glucose or triglyceride levels but caused a modest elevation of total plasma cholesterol. Respiration and oxidative phosphorylation of isolated liver mitochondria were not affected by any experimental diet. In pancreatic acinar cells, a high-fat diet caused a significant decrease of basal respiration by ~15%, but no effects were observed on the maximal rate of uncoupled respiration, mitochondrial membrane potential, or NAD(P)H autofluorescence. In these cells, a ketone body 3-hydroxybutyrate caused elevation of uncoupled respiration and NAD(P)H level irrespectively of the diet. Diets did not cause any change in sperm concentration, viability or motility. Surprisingly, in animals on a high-fat high-sugar diet, a significant increase in both basal and maximal respiration of sperm cells was observed. Collectively, these data show that while the elevated fat and sugar content in the diet does not cause significant obesity, no detrimental effects on mitochondria of the liver, pancreas, and sperm cells are observed.

Prooxidant and antioxidant processes in the liver homogenate of healthy and tumor-bearing mice under the action of thiazole derivatives

Ya. R. Shalai1*, M. V. Popovych1, S. M. Mandzynets1,
V. P. Hreniukh1, N. S. Finiuk1,2, A. M. Babsky1

1Ivan Franko National University of Lviv, Ukraine;
2Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
*e-mail: Yaryna.Shalay@lnu.edu.ua

Received: 12 October 2021; Accepted: 17 May 2021

Thiazole derivatives were shown to have toxic effects in vitro on cancer cells of different origin and can be considered as potentially antineoplastic, but their effect on the normal tissues needs to be studied. In this research the newly synthesized thiazole derivatives of N-(5-benzyl-1,3-thiazol-2-yl)-3,5-dimethyl-1-benzo-furan-2-carboxamide (BF1) and 8-methyl-2-Me-7-[trifluoromethyl-phenylmethyl]-pyrazolo [4,3-e] [1,3] thiazolo [3,2-a] pyrimidin-4 (2H)-one (PP2) were used and their effect on the pro- and antioxidant processes after adding in a 1, 10 and 50 μM concentrations to the liver homogenate of healthy and NK/Ly lymphoma-bearing mice was estimated. The level of superoxide radical and TBA-active products as well as catalase, SOD and glutathione peroxidase activity were measured. It was shown that superoxide radical and TBA-active products­ level and catalase activity were significantly higher in the liver of tumor-bearing mice than in the liver of the healthy mice. Neither BF1 no PP2 influenced the studied indices in the liver homogenate of healthy and tumor-bearing animals with the exception that PP2 significantly reduced the level of TBA-positive products in both cases. The data obtained showed that the studied thiazole derivatives did not cause severe liver toxicity in both healthy and tumor-bearing mice.

Effects of ethylthiosulfanylate and chromium (VI) on the state of pro/antioxidant system in rat liver

B. І. Kotyk1, R. Ya. Iskra1, O. M. Slivinska1, N. M. Liubas1,
A. Z. Pylypets1, V. I. Lubenets2, V. I. Pryimych3

1Institute of Animal Biology, NAAS of Ukraine, Lviv;
2Lviv Polytechnic National University, Ukraine;
3Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv, Ukraine;
e-mail: kicyniabo@gmail.com

Received: 04 April 2020; Accepted: 25 June 2020

Ethylthiosulfanylate is alkyl ester of thiosulfoacid and belongs to the class of thiosulfonate compounds. Structurally, thiosulfonates are synthetic analogues of natural phytoncides. It is known that, natural organic sulfur-containing compounds are characterized by antioxidant and detoxification properties against heavy metals toxicity. Therefore, the purpose of the study was to investigate the influence of ethylthiosulfanylate, as a synthetic analogue of natural phytoncides, on the state of the pro/antioxidant system in the liver of laboratory rats exposed to Cr(VI). It was found that ethylthiosulfanylate exposure at a dose 100 mg/kg body weight daily for 14 days led to a decrease in the intensity of increasing of the lipid hydroperoxides (LHP) content in the rat liver caused by Cr(VI) action. In addition, ethylthiosulfanylate pretreatment prevented depletion of reduced glutathione (GSH) pool under the action of potassium dichromate oxidative stress and performed the accumulation of cellular GSH in rat liver.

Vitamin D(3) regulates hepatic VEGF-A and apelin expression in experimental type 1 diabetes

D. O. Labudzynskyi1*, I. O. Shymanskyi1, O. O. Lisakovska1,
A. O. Mazanova1, L. V. Natrus2, M. M. Veliky1

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Bogomolets National Medical University, Kyiv, Ukraine;
*e-mail: konsument3@gmail.com

Received: 09 July 2019; Accepted: 15 May 2020

The deficiency of vitamin D is associated with the risk of various chronic diseases, including diabetes mellitus and its complications. Given the strong genomic action of vitamin D hormone-active form, its deficiency can lead to dysfunction of cytokine signaling pathways, including those dependent on vascular endothelial growth factors (VEGFs) and apelin. The present study was carried out to define the link between VEGF-A and apelin expression in liver, hepatocytes viability and vitamin D status at experimental type 1 diabetes in mice. We established that chronic hyperglycemia at streptozotocin-induced diabetes was accompanied by a 2.2-fold decrease in 25OHD content in the serum and increased hepatocytes apoptosis and necrosis. Vitamin D deficiency correlated with increased apelin and VEGF-A (8- and 1.6-fold respectively) expression. Almost complete restoration of circulatory 25OHD content in serum was achieved at vitamin D3 treatment (800 IU/kg, per os, for 2 months) followed by reduced apelin and VEGF-A expression in liver and the decline of hepatocytes apoptosis. We conclude that vitamin D3 can be involved in cell survival, angiogenesis and fibrogenesis by modulating  VEGF-A and apelin dependent regulatory systems in diabetic liver.

Corvitin modulates the content of lipids in rat liver bile

T. V. Vovkun1, P. I. Yanchuk1, L. Ya. Shtanova1,
S. P. Veselsky1, N. B. Filimonova1, I. V. Komarov2

1ESC “Institute of Biology and Мedicine”, National Taras Shevchenko University of Kyiv, Ukraine
2“Institute of High Technologies
”, National Taras Shevchenko University of Kyiv, Ukraine;
e-mail: shtanova@ukr.net

Received: 23 March 2019; Accepted: 18 October 2019

Quercetin (QUE) and its water-soluble form сorvitin are medicinally important members of the fla­vonoid family and the most prominent dietary antioxidants. Numerous pharmacological effects of quercetin include protection against diseases, such as atherosclerosis, myocardial infarction, and cerebrovascular disea­ses. Corvitin modulates liver blood flow but its effects on liver lipid metabolism have not been understood. We investigated the influence of corvitin (2.5, 5, 10 mg/kg) on the formation and secretion of cholesterol (Chol), cholesterol esters (EChol), phospholipids (PLs), free fatty acids (FFAs), and triglycerides (TGs) into the bile. Secreted bile was collected during 2.5 h of the experiment. Lipid fractions of the bile were separated by thin-layer chromatography. We defined that administration of corvitin caused a significant (P < 0.05–0.001) increase in levels of all studied lipid components of bile. At a dose of 2.5 and 5 mg/kg corvitin increased bile content of Chol, PLs and FFAs,and at a  dose of 10 mg/kg had the largest effect on the total production of EChol and TGs. We concluded that corvitin activates the liver lipids metabolism and processes of bile formation.

Long-term hypocholesterolemic effect of amidated alginate in rats

M. Marounek1, Z. Volek1, T. Taubner1, D. Dušková1, L. Kalachniuk2

1Institute of Animal Science, Prague, Czech Republic;
e-mail: marounek.milan@vuzv.cz;
2National University of Life and Environmental Sciences of Ukraine, Kyiv;
e-mail: kalachnyuk_liliya@nubip.edu.ua; lilkalachnyuk@gmail.com

The effect of octadecylamide of alginic acid on blood serum and hepatic cholesterol, and the faecal output of fat and sterols was examined in female rats fed diets containing cholesterol and palm fat at 10 and 50 g/kg, respectively for 10 weeks. Amidated alginate, supplied at 10 and 20 g/kg, significantly decreased serum cholesterol from 5.25 to 2.99 and 2.39 µmol/ml, respectively, and decreased hepatic cholesterol from 30.7 to 12.3 and 9.4 µmol/g, respectively. Amidated alginate increased the faecal output of fat and at higher dosing significantly decreased faecal output of bile acids. Faecal output of bile acids and hepatic cholesterol significantly correlated (r = 0.791; P < 0.001). The results of the present experiment showed that hypocholesterolemic effect of amidated alginate persisted within 10 weeks of feeding.

Liver cytochrome P450-hydroxylation system of tumor-bearing rats under the influence of ω-3 polyunsaturated fatty acids and vitamin D(3)

I. O. Shymanskyi1, O. V. Ketsa2, M. M. Marchenko2, М. М. Veliky1

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: ihorshym@gmail.com;
2Fedkovich Chernovtsy National University, Chernovtsy

The study was performed to investigate the effects of the separate and combined action of omega-3 polyunsaturated fatty acids (ω-3 PUFAs) and vitamin D3 on the activity of the components of the oxygenase and reductase chains of the monooxygenase system (MOS) in the microsomal fraction isolated from the liver of rats with transplanted Guerin carcinoma. In the liver of the tumor-bearing rats during the intensive growth of the tumor (14 days, which corresponds to the logarithmic phase of tumor growth), the functional activity of the MOS was weakened. N-demethylase, p-hydroxylase and NADPH-cytochrome P450 reductase activity decreased, with the simultaneous enhancement of cytochrome P450 inactivation rate due to its transformation into an inactive form, cytochrome P420. In turn, we found an increase in the functional activity of the reductase chain of MOS, which components are known to transfer electrons from the reduced NADH through NADH-cytochrome b5-reductase and cytochrome b5 to cytochrome P450. In particular, the activity of NADH-cytochrome b5-reductase and the rate of reduction of cytochrome b5 were elevated with a simultaneous decrease in its content. Both ω-3 PUFAs and vitamin D3 administration to tumor-bearing rats for 42 days (28 days of preliminary administration and 14 days of tumor growth) significantly normalized the oxygenase activity of MOS, increasing NADPH-cytochrome P450-reductase, N-demethylase and p-hydroxylase activity of cytochrome P450 and blocking cytochrome P450 inactivation rate in the microsomal fraction of the liver. Administration of ω-3 PUFAs in combination with vitamin D3 led to the synergy. Changes in the activity of the components of the reductase chain of MOS in liver of tumor-bearing rats were observed mainly after ω-3 PUFAs supplementation. The content of cytochrome b5 increased and the rate of its reduction was significantly diminished. In the absence of a pronounced individual effect of vitamin D3 on the reductase chain of MOS, its co-administration with ω-3 PUFA was also found to be ineffective.