Tag Archives: liver

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.

The relationship between the ionized Ca concentration and mitochondrial functions

L. G. Babich1, S. G. Shlykov1, A. M. Kushnarova-Vakal1, N. I. Kupynyak2, V. V. Manko2, V. P. Fomin3, S. O. Kosterin1

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: babich@biochem.kiev.ua;
2Ivan Franko National University of Lviv, Ukraine;
3University of Delaware, Newark, USA

The aim of the study was to show the relationships between ionized Ca concentration ([Ca2+]m) in the mitochondria matrix and functional activity of this organelle. [Ca2+]m was determined using the fluorescent probe Fluo-4, AM. Total level of Ca2+ accumulation in mitochondria was monitored using 45Ca2+ as radioactive tracer. It was shown that incubation of myometrium mitochondria with 3 mM Mg2+ resulted in the low level of [Ca2+]m. Subsequent addition of 100 µM Ca2+ resulted in 8 times increase of [Ca2+]m but in low level of total calcium accumulation. Normalized fluorescence of Ca2+-sensitive probe Fluo-4 in response to the Ca2+ addition was higher than 2.5. At the same time, [Ca2+]m was considerably higher in the medium containing­ 3 mМ АТР and 3 mМ Mg2+. Subsequent addition of 100 µM Ca2+ to the incubation medium resulted in only 2.4 times increase of [Ca2+]m but considerably higher level of total calcium accumulation was observed. Normali­zed fluorescence of Fluo-4 in response to the Ca2+ addition was lower than 1.3. In liver mitochondria higher rate of oxygen consumption was detected in the presence of an oxidative substrate succinate than of pyruvate or α-ketoglutarate. At the presence of an oxidative substrate succinate normalized fluorescence of Fluo-4 in liver mitochondria in response to the Ca2+ addition was lower than 1.3. It was concluded that low level of [Ca2+]m was correlated with low functional activity of this organelle and, vise versa, high level of [Ca2+]m was correlated with high functional activity. It was suggested that normalized fluorescence changes in response to the Са2+ addition could be used as a test of the mitochondrial functional activity: lower normalized fluorescence values − higher functional activity.

Metal-binding capacity of metallothioneins of the liver of rats poisoned with heavy metals

B. O. Tsudzevich, I. V. Kalinin

Taras Shevchenko Kyiv National University, Ukraine;
е-mail: ikalin@rambler.ru

The functioning of metallothioneins in the liver of rats, poisoned with copper sulfate and cadmium sulfate has been investigated. By sequential chromatography on sephadex G-50 and DEAE-cellulose the authors obtained metallothioneins (MT-1, MT-1A, MT-2, MT-2a), which differ in molecular weight and composition of associated metals. Heavy metal poisoning leads to activation of synthesis and metal-binding function of metallothioneins, as well as to changes in the composition of their isoforms.

The liver and kidneys biochemical indices at the experimental pancreatitis in case of the administration of nitric oxide synthesis modulators and recombinant superoxide dismutase

O. Z. Yaremchuk, K. A. Posokhova

SHEI «I. Ya. Horbachevsky Ternopil State Medical University», Ukraine;
e-mail: yaremchukolya@rambler.ru

The rats liver and kidneys function indices were studied in case of administration of recombinant superoxide dismutase drug, precursor of nitric oxide L-arginine and the blocker of inducible NO-synthase aminoguanidine. The disturbances in functioning of prooxidant-antioxidant system (a decrease of activity of superoxide dismutase, katalaze, amount of restored glutathione, growth of the level of hydroperoxide lipids, TBA-active products­), mitochondrial electron-transport pathways (a decrease in activity of succinatedehydrogenaze, cytochrome oxydaze), a rise of nitrite-anion level in the liver and kidneys, increase of α-amylase activity and tumor necrosis factor α serum concentration were established on the model of pancreas injury in white male rats. Under these circumstances aminoguanidine attenuated the oxidative stress in the liver and kidneys due to normalization of nitric oxide synthesis. The ability to activate the antioxidant system was proved by combined usage of recombinant superoxide dismutase and aminoguanidine. It was determined that recombinant superoxide dismutase partially decreases the negative influence of L-arginine and improves the biochemical indices of the liver and kidneys function in rats with acute experimental pancreatitis.

Effects of thiosulfonates on the lipid composition of rat tissues

A. Z. Pylypets1,2, R. Ya. Iskra2, V. V. Havryliak1,2, A. V. Nakonechna1, V. P. Novikov1, V. I. Lubenets1

1Lviv Polytechnic National University, Ukraine;
2Institute of Animal Biology, NAAS of Ukraine, Lviv;
e-mail: vlubenets@gmail.com

Thiosulfonates are synthetic analogs of organic sulfur-containing compounds isolated from plants. Recent studies have shown that these substances lowering cholesterol content in the body, are effective against hyperlipidemia. Therefore, the aim of our investigation was to study the effect of synthesized thiosulfonates on the content of lipids and their spectrum in rats blood, liver and kidney. The amount of total lipids and their fractional profile were determined by thin-layer chromatography. The administration of methyl-, ethyl-, and allylthiosulfonates at a dose of 300 mg/kg of body weight did not cause significant changes in the content of total lipids and phospholipids, but led to the redistribution of their classes in the examined tissues. The content of triacylglycerols in the blood plasma under the action of ethyl- and allylthiosulfonates was decreased by 29.14 and 23.19% (P < 0.05-0.01), respectively, whereas the injection with methyl- and ethylthiosulfonates was accompanied by a significant decrease in mono-, di-, triglycerides and free fatty acids in the liver compared to control. The most significant changes in the lipid profile of kidney tissue were detected under the action of methylthiosulfonate.

Influence of antitumor system rhenium–platinum on biochemical state of the liver

V. V. Ivchuk1, T. N. Polishko1, O. A. Golichenko2,
O. V. Shtemenko2, N. I. Shtemenko1

1Oles’ Gonchar Dnepropetrovsk National University, Ukraine;
2Ukrainian State University of Chemical Technology, Dnepropetrovsk;
e-mail: n.shtemenko@i.ua

Influence of the antitumour rhenium-platinum system on biochemical liver characteristics in the model of tumor growth (Guerin carcinoma) was studied and possible hepatoprotective activi­ty of rhenium cluster compounds when introducing them in different forms was shown, that was confirmed by decreasing of diagnostic enzymes activity in blood (aminotransferase – AST 6 times and ALT 5.6 times, lactatedehydrogenase 4.9 times, γ-glutamyltranspeptidase 3.6 times) and normalization of morphological state of the liver cells. The hepatoprotective activity of the cluster rhenium compound with adamanthyl ligands was confirmed in the model of acute toxic hepatitis. Introduction of this compound led to reduction of the concentration of MDA in homogenates of liver tissue (2 times), and in blood plasma (3.8 times); to reduction of levels of diagnostic liver­ enzymes in blood – AST and ALT 5.8 and 5.5 times respectively in comparison with control group. Some aspects of the mechanism of hepatoprotection were discussed, that included the presence of conjugated systems around the quadrupol rhenium-rhenium bond and alkyl radicals with significant positive inductive effects.

Generation of superoxide anion-radical in the liver monooxygenase system of preliminary radiation-exposed tumor-bearing rats

М. М. Marchenko, O. V. Ketsa

Yuriy Fedkovych Chernivtsi National University, Ukraine;
e-mail: ketsa80@mail.ru

Generation of superoxide anion-radical (O2•-) in reductase and oxygenase electron-transport chains in the liver monooxygenase system was investigated in tumor-bearing rats exposed to preliminary irradiation. Preliminary irradiation of rats (before transplantation of Guerin’s carcinoma) resulted in the increased generation of superoxide anion-radical by components of the liver monooxygenase system in the logarithmic phase of oncogenesis­.
It is shown that the increased NADPH-cytochrome P-450 reductase activity is accompanied with the intensification of superoxide anion-radical generation in liver microsomal fraction of preliminary radiation-exposed rats. At the same time the cytochrome P-450 hydroxylase activity in the microsomal fraction of tumor-bearing rats subjected to preliminary irradiation was decreased.

Antitoxical effects of N-stearoylethanolamine in suspension and in nanocomposite complex in the organs of mice with the Lewis carcinoma under doxorubicin administration

I. A. Goudz1, N. M. Gula1, T. O. Khmel1, T. M. Goridko1, Y. M. Bashta1,
R. R. Panchuk2, R. S. Stoika2, A. A. Ryabtseva3, O. S. Zaichenko3

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: ngula@biochem.kiev.ua;
2Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
3National University Lviv Politekhnika, Ukraine

The antioxidant effects of N-stearoylethanolamine (NSE) in the nanocomplex composition and in suspension are shown on the model of intoxication by doxorubicin in conditions of development of the Lewis carcinoma in the heart, kidneys and liver tissue and in the blood plasma of female mice. The NSE suspension reduces the level of urea in the blood plasma of mice with the Lewis carcinoma, which growth was revealed as a result of introduction of doxorubicin. Under introduction of nanocomplex the amount of urea remains at the level of that in the intact mice. In the blood plasma of mice with the Lewis carcinoma the NSE suspension and nanocomplex reduce activity of aspartate aminotransferase, the basic marker of necrosis of the heart tissue, growth of which was caused by the tumour development. Doxorubicinum increases activity of alanine aminotransferase, the marker of the liver lesion; introduction of NSE in the nanocomplex composition prevents the growth of the enzyme activity. N-stearoylethanolamine, both in the nanocomplex and in suspension, modulates activity of enzymes of antioxidantive protection of the heart, kidney and liver tissue of mice with the Lewis carcinoma.

Activity of enzymatic detoxification systems in the mice liver under conditions of different retinoid provision

М. M. Marchenko, G. P. Kopylchuk, I. O. Shmarakov, I. M. Buchkovska

Yuriy Fedkovych Chernivtsi National University, Ukraine;
e-mail: ivannabuchkovska@mail.ru

The activity of cellular components of liver detoxification system was studied under the conditions of the absence of vitamin A stores. It is shown that a decrease of p-hydroxylase and N-demethylase activity of cytochrome P-450 simultaneously with a decrease of glutathione-S-transferase activi­ty takes place in the liver microsomal fraction of vitamin A-deficient animals. At the same time the absence of retinoid stores in knock-out animals influences the decrease of only p-hydroxylase activity of cytochrome P-450 system. The increase in glutathione-S-transferase activity is observed in the liver postmicrosomal fraction in mice, kept on vitamin A-deficient diet, while its parametres in knock-out group animals were not statistically different compared to the control.

Alterations of prooxidant-antioxidant system of rat liver at ethanol and tetracycline action

Kh. Yu. Nedoshytko

I. Ya. Horbachevsky Ternopil State Medical University, Ukraine;
e-mail: khrystynan@ukr.net

The state of antioxidant system and fatty acid composition of lipids in the liver tissues of rats of different sex at the ethanol and tetracycline action and at the influence of biologically active additives (BAA) “Alpha + Omega” at a dose of 0.5 mg/kg b.w. per os was investigated. It was found that the contet of lipid peroxidation products in the liver was increased at the action of 40 % ethanol at a dose of 7 ml/kg b.w. per os and tetracycline – 500 mg/kg and more profound at their joint using. Howe­ver, the content of diene conjugates was stronger increased in the liver of females at the action of ethanol, while in the liver of males at the action of tetracycline (Р < 0.05). It was shown that the application of the investigated compounds led to the reduction of an antioxidant defense system activity of males and females liver, as evidenced by the decrease of superoxide dismutase activity by 46 and 43% and reduction of glutathione content by 39 and 38% (Р < 0.05). The activity of alanineaminotransferase, aspartateaminotransferase and alkalinephosphatase was increased in the liver of males and females under the influence of ethanol and tetracycline and more profound at their joint usa­ge (Р < 0.05). It was established that ethanol and tetracycline unidirectionally changed fatty acid composition of total lipids of rat liver, but at the ethanol action the changes were more expressed in females while at the tetracycline action in males. The application during 14 days of BAA “Alpha + Omega” to male and female rats with an acute tetracycline damage at subacute ethanol action led to partial normalization of prooxidant-antioxidant system and the relative content of total lipids fatty acids of the liver of both sexes animals.