Tag Archives: mitochondria
Curcumin exerts protective effects against doxorubicin-induced cardiotoxicity
O. О. Klymenko1*, T. I. Drevytska1, O. O. Gonchar1, K. V. Tarasova2,
V. I. Nosar1, V. Ye. Dosenko1, I. M. Mankovska1
1Bogomoletz Institute of Physiology, National Academy of Sciences of Ukraine, Kyiv;
2Bogomolets National Medical University, Kyiv, Ukraine;
*e-mail: klymenkooks2018@gmail.com
Received: 16 October 2024; Revised: 03 December 2024;
Accepted: 21 February 2025; Available on-line: 03 Marchy 2025
The mechanism of doxorubicin (Dox) cardiotoxicity involves different pathways, including oxidative stress and mitochondrial dysfunction. It’s supposed that pharmacological effect on HIF gene expression may protect the heart against the detrimental effects of the doxorubicin-induced injury. We hypothesized that the cardioprotective effects of Curcumin (Curc) are exerted by regulating HIF and its target genes expression. To test this, an in vitro model of Dox-induced injury to primary myocardial cardiomyocytes was used. Isolated Wistar rat neonatal cardiomyocytes were incubated in the culture medium for 24 h in control, either with Dox (0.5 μmol/ml) or Curc (20 μmol/ml), or in their combination in the same doses. Mitochondria were isolated from rat cardiomyocytes culture. It was demonstrated that cardiomyocytes exposure to Dox led to an increase in the activity of oxidative stress markers in isolated mitochondria, a decrease in the efficiency of the respiratory chain and phosphorylation processes, decline of membrane potential and the rate of K+ ions entry into mitochondria. Doxorubicin inhibited the expression of mRNA of both HIF-1α, 2α, 3α subunits and its important target genes PDK-1 and IGF-1 in mitochondria. A negative impact on the cardiomyocyte contractile activity was observed. The combined use of doxorubicin with curcumin led to an increase of cardiomyocytes viability and attenuation of oxidative stress in mitochondria, prevented the development of mitochondrial dysfunction and significantly improved the contractile activity of cardiomyocytes.
Thiacalix[4]arene chalcone amides effect on myometrium contraction
O. V. Tsymbalyuk1*, S. G. Shlykov2, L. G. Babich2, О. Yu. Chunikhin2,
R. V. Rodik3, S. G. Vyshnevskyi3, O. A. Yesypenko3, S. O. Kosterin2
1Educational and Scientific Institute of High Technologies,
Taras Shevchenko National University of Kyiv, Ukraine;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
3Institute of Organic Chemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: otsymbal@knu.ua
Received: 26 April 2024; Revised: 26 June 2024;
Accepted: 25 July 2024; Available on-line: 04 September 2024
Calixarenes are macrocyclic compounds, the biochemical effects of which are actively studied. In this study we synthesized thiacalix[4]arene chalcone amides С-1191 and С-1192, which have a sulfur atom in their structure and different spatial arrangement of chalcone amide groups, and studied their effect on myometrium functioning. Experiments were conducted with the use of rat uterine smooth muscles preparations, isolated myometrial mitochondria and permeabilized myometrial cells. The relative value of mitochondria membrane potential (Δψ) was assayed with a voltage-sensitive fluorescent probe TMRM. The spontaneous contractive activity was studied by tenzometric method followed by mechanokinetic analysis. It was shown that C-1191 and C-1192 induced mitochondria hyperpolarization and increased the basal tension of myometrium smooth muscle preparation. Thiacalix[4]arene С-1191 did not change the uterine cycle, but increased the force, velocity and impulse parameters of muscle contractive activity. On the contrary, С-1192 modified the uterine cycle considerably, increased the total efficiency of the myometrium spontaneous contractive activity and decreased the force, time and impulse parameters. It is concluded that changes in the mechanokinetic parameters of myometrial contractile activity induced by С-1191 and С-1192 are determined by the functional activity of mitochondria.
ATP as a signaling molecule
L. G. Babich*, S. G. Shlykov, S. O. Kosterin
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: babich@biochem.kiev.ua
Received: 22 January 2024; Revised: 13 February 2024;
Accepted: 31 May 2024; Available on-line: 17 June 2024
The review considers the effects of extracellular ATP mediated by plasma membrane purinoreceptors in the cells of different tissues, in particular, myometrium. Recently published results suggest that cytosolic ATP may also play a role of signaling molecule, as indicated by the detection of the ATP receptor not only in the plasma membrane, but also in mitochondria. The authors have shown that ionized Ca2+ concentration in the rat myometrium mitochondria matrix is regulated by ATP at the absence of exogenous Ca2+. ATP concentration-dependent increase of [Ca2+]m was not affected in the presence of the mitochondrial Ca2+-uniporter blocker ruthenium red, the mitochondrial pore blocker cyclosporine A, or ATP synthase inhibitor oligomycin. It is assumed that cytosolic ATP could be a signaling molecule that regulates at least the Ca2+ ions exchange in mitochondria.
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.
Choline derivatives as natural ligands of mitochondrial nicotinic acetylcholine receptors
O. Lykhmus, M. Izmailov, M. Skok*
Department of Molecular Immunology, Palladin Institute of Biochemistry,
National Academy of Sciences of Ukraine, Kyiv;
*e-mail: skok@biochem.kiev.ua
Received: 16 March 2023; Revised: 28 April 2023;
Accepted: 05 June 2023; Available on-line: 20 June 2023
Nicotinic acetylcholine receptors (nAChRs) regulate mitochondria-driven apoptosis; however, their intracellular ligands are unknown. In the present paper, we show that choline and its derivatives (phosphocholine (PC), L-α-glycerophosphocholine (G-PC) and 1-palmitoyl-sn-glycero-3-phosphocholine (P-GPC)) dose-dependently influence cytochrome c release from isolated mouse liver mitochondria. Choline inhibited Ca2+-stimulated cytochrome c release, while PC attenuated wortmannin-induced cytochrome c release. Small doses of G-PC and P-GPC (up to 0.1 µM) were protective against either Ca2+ or wortmannin, while larger doses (up to 1 µM) stimulated cytochrome c release by themselves. Choline and PC disrupted interaction of VDAC1, Bax and Bcl-2 with mitochondrial α7 nAChRs and favored their interaction with α9 nAChR subunits. It is concluded that choline metabolites can regulate apoptosis by affecting mitochondrial nAChRs.
Biochemical and molecular-physiological aspects of the nitric oxide action in the utera
H. V. Danylovych, Yu. V. Danylovych
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: danylovych@biochem.kiev.ua
Received: 18 May 2021; Accepted: 12 November 2021
The sources of the nitric oxide (NO) formation in the uterus and the dynamics of changes in its content in different periods of organ functioning in human and animals are analyzed. The biochemical mechanisms of NO action on the myometrium contractile activity, the significance of NO in the physiological processes during pregnancy and labor, the importance of mitochondria as a reliable NO source in the smooth muscle and the possible ways of NO influence on Ca2+ transport and bioenergetic processes in mitochondria are considered. The authors’ data concerning ionic and membrane mechanisms of NO action on Ca2+-homeostasis of uterine myocytes, identification of nitric oxide in uterine smooth muscle mitochondria, biochemical characteristics of the NO-synthase reaction and the possible role of NO in the regulation of Ca2+ transport in these subcellular structures and in the electron transport chain functioning are presented and discussed.
Application of petri nets methodology to determine biophysicochemical parameters of mitochondria functioning
H. V. Danylovych*, A. Yu. Chunikhin, Yu. V. Danylovych, S. O. Kosterin
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: danylovych@biochem.kiev.ua
Received: 01 Nobember 2020; Accepted: 17 May 2021
With the use of Petri net methodology a mathematical simulation model able to predict simultaneous changes in biophysicochemical parameters of mitochondria functioning was developed. The model allowed to interconnect in time the changes in mitochondria hydrodynamic diameter, electronic transport chain functioning, endogenous fluorescence of adenine nucleotides, DCF fluorescence signal of ROS production and NaN3 effects. It was shown that the calculated values of the studied biophysicalchemical parameters correspond to those obtained experimentally. The model permit to link mitochondrial functional changes and their structural representation and to optimize significantly experimental procedures.
Effects of thiazole derivatives on intracellular structure and functions in murine lymphoma cells
V. P. Hreniukh1, N. S. Finiuk1,2, Ya. R. Shalai1, B. O. Manko1,
B. V. Manko1, Yu. V. Ostapiuk1, O. R. Kulachkovskyy1,
M. D. Obushak1, R. S. Stoika1,2, A. M. Babsky1*
1Ivan Franko National University of Lviv, Ukraine;
2Institute of Cell Biology, Nationl Academy of Sciences of Ukraine, Lviv;
*e-mail: andriy.babsky@gmail.com
Received: 22 December 2019; Accepted: 27 March 2020
Thiazole derivatives have cytotoxic effects towards tumor cells, such as glioblastoma, melanoma, leukemia and lymphoma. However, the intracellular mechanism of this action is not clear. The aim of our study was to investigate the action of N-(5-benzyl-1,3-thiazol-2-yl)-3,5-dimethyl-1-benzofuran-2-carboxamide (BF1) and 7-benzyl-8-methyl-2-propylpyrazolo[4,3-e]thiazolo[3,2-a]pyrimidin-4(2H)-one (PP2) on cellular structure, and bioenergetic functions of mitochondria in Nemeth-Kellner lymphoma cells (NK/Ly). The structure of treated NK/Ly cells and their mitochondria was examined using electron microscopy. The rate of oxygen uptake by isolated mitochondria was recorded by a polarographic method using a Clark electrode. The mitochondrial potential relative values were registered using fluorescence dye rhodamine 123. In the short-term (15 min), incubation with BF1 and PP2 in 10 and 50 µM concentrations induced apoptotic and necrotic changes in the structure of NK/Ly cells, such as fragmentation and disintegration of the nucleus, destruction of the plasma membrane, and an increase in numbers of lysosomes and mitochondria. A polarographic method did not show significant metabolic shifts in lymphoma mitochondria, in either in vitro or ex vivo actions of the thiazole derivatives. However, fluorescent microscopy showed a significant decrease in mitochondria potential, following a 15 min incubation of cells with 50 µM of PP2. Thus, the electron and fluorescent microscopy data suggest that mitochondria are involved in the mechanism of cytotoxic action of the studied thiazole derivatives.
Sources and regulation of nitric oxide synthesis in uterus smooth muscle cells
H. V. Danylovych, Yu. V. Danylovych, T. V. Bohach,
V. T. Hurska, S. O. Kosterin
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: danylovych@biochem.kiev.ua
Received: 28 February 2019; Accepted: 17 May 2019
It was proved that NO synthesis in isolated mitochondria of rat uterus smooth muscle depended on the entry of exogenous Ca2+ to mitochondria (inhibited by 1-10 mM Mg2+ in the absence of ATP and by 10 μM ruthenium red) and was suppressed by calmodulin antagonists (0.1-10 μM calmidazolium and 1-100 μM trifluoperazine). It was blocked by NG-nitro-L-arginine, a known antagonist of the constitutive NO-synthase, with a half-maximal inhibition effect at about 25 μM. Moderate deholesterinization of the plasma membrane of myocytes after processing with 0.01% digitonin was followed by increased nitric oxide biosynthesis by cells. The data obtained suggested that mitochondria and plasmalemma is a possible source of NO synthesis in uterine myocytes.