Category Archives: Uncategorized
NO-synthase activity and nitric oxide content in lymphoid cells of thymus and spleen of rats under conditions of diet-induced obesity
L. Kot*, V. Konopelnyuk, K. Dvorshchenko, V. Vereschaka
ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine;
*e-mail: kot_lora@ukr.net
Received: 10 August 2020; Accepted: 17 May 2021
Metabolic disorders under conditions of obesity are known to be accompanied by systemic inflammation and immune system imbalance while nitric oxide (NO) system is one of the main key links in the immune responses regulation. The aim of the research was to evaluate NO-synthase (NOS) activity and NO content in the cells of thymus and spleen of rats with experimental obesity induced by 14 weeks of high-calorie diet. NO content in cell suspensions was determined using Griess reaction, NOS activity was estimated by NADPH++Н+ specific cleavage. It was shown that under conditions of experimental obesity NO content in cells of thymus and spleen as well as NOS activity in the spleen were decreased with simultaneous increase in both NO content and NOS activity in the serum. The obtained data testify the changes in lymphoid cells functional activity and in organism general adaptive capacity under conditions of obesity.
Sex dependent differences in oxidative stress in the heart of rats with type 2 diabetes
N. I. Gorbenko1*, O. Yu. Borikov2, O. V. Ivanova1, T. V. Kiprych1,
E. V. Taran1, T. I. Gopciy2, Т. S. Litvinova1
1SI “V. Danilevsky Institute for Endocrine Pathology Problems of the NAMS of Ukraine”, Kharkiv;
2V.N. Karazin Kharkiv National University, Kharkov, Ukraine;.
*е-mail: Gorbenkonat58@ukr.net
Received: 17 September 2020; Accepted: 17 May 2021
Type 2 diabetes mellitus is known to double mortality from cardiovascular diseases (CVD), in which oxidative stress plays an important role. It is suggested that the impact of diabetes on CVD risk may vary depending on gender. The aim of the study was to assess oxidative stress parameters in the heart of 12 weeks old male and female Wistar rats with type 2 diabetes mellitus (T2DM) induced by high-calorie diet followed by intraperitoneal streptozotocin injections. The level of advanced oxidation protein products, superoxide dismutase, glutathione reductase and glutathione peroxidase activity in the isolated heart mitochondria and NADPH-oxidase and xanthine oxidase activity in the post-mitochondrial supernatant fraction were determined. It was shown that T2DM induced more pronounced oxidative stress confirmed by the increased level of advanced oxidation protein products in the heart mitochondria of males than females. The data obtained indicate that the main reason of oxidative stress in the heart of diabetic males is the activation of non-mitochondrial sources of reactive oxygen species. While in the heart of diabetic female rats it is the decrease in antioxidant enzymes activity in mitochondria. These results justify the necessity of gender-specific therapy for the prevention and management of diabetic CVD.
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.
Comparative cytotoxic activity of carboplatin and β-cryptoxanthin in free and liposomal forms against breast cancer cell line
M. W. Shafaa*, N. S. Elkholy
Physics Department, Medical Biophysics Division, Faculty of Science,
Helwan University, Cairo, Egypt;
*e-mail: medhatwi@hotmail.com
Received: 1 June 2020; Accepted: 17 May 2021
The study of the effectiveness of the synthetic and natural anticarcinogenic compounds in liposomal form is urgent for their possible use in therapy. In this work, the alkylating agent carboplatin and the representative of carotenoids β-cryptoxanthin were used. The aim of the research was to study the toxicity of these compounds in free and liposomal forms against breast cancer MCF-7 cell line. According to DSC and FTIR data, when carboplatin or β-cryptoxanthin were added to liposomal bilayers, a single peak was observed indicating their mutual mixing. Integration of β-cryptoxanthin into bilayer was found to be more proper for the creation of PE acyl chains ordered and cooperative state. It was found that MCF-7 cells sensitivity was much higher to the free β-cryptoxanthin than to the free carboplatin with IC50 42 and 235 μg/ml, respectively. The IC50 values for β-cryptoxanthin loaded into liposomes and for free carboplatin were similar. At the same time, no cytotoxic effect of carboplatin-loaded liposomes was observed. The data obtained allow proposing a possible antitumor treatment regimen where carboplatin is replaced by free β-cryptoxanthin or its liposomal form to increase the effectiveness of breast cancer therapy.
The effect of CO donor hemin on the antioxidant and osmoprotective systems state in Arabidopsis of a wild-type and mutants defective in jasmonate signaling under salt stress
M. A. Shkliarevskyi1, Yu. E. Kolupaev1,2*, T. O. Yastreb1,
Yu. V. Karpets1, A. P. Dmitriev3
1Dokuchaev Kharkiv National Agrarian University, Ukraine;
2Karazin Kharkiv National University, Ukraine;
3Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: plant.biology.knau@gmail.com
Received: 6 December 2020; Accepted: 17 May 2021
The role of the gasotransmitter carbon monoxide (CO) in signaling and adaptive processes in plants has been studied insufficiently. There are indirect data indicating jasmonate signaling participation in realization of CO effects, but the possible connection between carbon monoxide and jasmonate signaling during plant adaptation to salt stress remains unclear. We studied the carbon monoxide donor hemin effect on the Arabidopsis of a wild-type (Col-0) and defective in jasmonate signaling coi1 and jin1 mutants response to the salt stress. Arabidopsis thaliana 4-week-old plants were grown on a modified Hoagland’s medium. Plants were incubated for 24 h in usual or 2 µM hemin containing culture medium, then transferred to 150 mM NaCl containing media and incubated for 24 h before the medium was replaced with the usual one. It was shown that salt stress caused water deficiency and superoxide dismutase and catalase activity decrease in the plants of all three genotypes. Treatment with 2 μM hemin stabilized the levels of catalase activity and photosynthetic pigments and increased guaiacol peroxidase activity in a wild-type, but not in coi1 and jin1 mutant plants after stress induction. Treated with hemin wild-type Arabidopsis plants accumulated more proline and sugars in response to stress than treated coi1 and jin1 mutants. It was concluded that jasmonate signaling can be involved in adaptive processes induced by exogenous carbon monoxide.
Administration of platelet-rich plasma or concentrated bone marrow aspirate after mechanically induced ischemia improves biochemical parameters in skeletal muscle
А. Т. Pidlisetsky1, G. V. Kosiakova2, T. M. Goridko2, A. G. Berdyschev2,
O. F. Meged2, S. I. Savosko3*, О. V. Dolgopolov4
1Lviv Regional Hospital for War Veterans and Repressed named after Yu. Lypa, Ukraine;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
3Bogomolets National Medical University, Kyiv, Ukraine;
4SI “Institute of Traumatology and Orthopedics of NAMS of Ukraine”, Kyiv;
*e-mail: savosko_s@ukr.net
Received: 16 September 2020; Accepted: 17 May 2021
Ischemia is known to cause structural and functional damage to skeletal muscle. In this research platelet-rich plasma (PRP) and cell bone marrow aspirate (CBMA) which are considered to be promising in regenerative medicine were used to improve the animal muscles condition after ischemia. Mechanically induced ischemia (MII) of the rabbit skeletal muscles was caused by applying a tourniquet to the hind limb for 6 hours. At 6 h after tourniquet was removed the rabbit PRP or CBMA solutions were injected into the tibia muscle and animals were divided into experimental groups (n = 15): 1 – ischemia + saline, 2 – ischemia + PRP, 3 – ischemia + CBMA. Intact animals were the control group (n = 5). Animals were removed from the experiment on 5, 15, and 30th days of reperfusion after ischemia. A progressive hypotrophy, muscle fibers destruction and leukocyte infiltration was observed after tibia muscle MII. The ischemic state was accompanied by catalase and superoxide dismutase activation, persistent NO2– overproduction and a significant decrease in glutathione peroxidase activity in the muscle homogenates. It was shown that after PRP or CBMA local injection into the ischemic muscle catalase activity remained at the elevated level, SOD activity approached the control level and the most pronounced effects were NO2– level decrease and glutathioperoxidase activity restoration. After CBMA injection angiogenesis activation and the increase in the number of nuclei in the muscle fibrils were observed. The obtained data allow to expand the range of biochemical parameters for assessing the muscles condition after ischemia and indicate a positive effect of platelet-rich plasma and cell bone marrow aspirate injection on skeletal muscle regeneration.
Lipid peroxidation and DNA fragmentation in fresh and cryopreserved spermatozoa of men at different spermatogenesis state
T. O. Yurchuk*, O. V. Pavlovich, G. O. Gapon,
A. Y. Pugovkin, M. P. Petrushko
Institute for Problems of Cryobiology and Cryomedicine, National Academy of Sciences of Ukraine, Department of Cryobiology of Reproductive System, Kharkiv;
*e-mail: taisiya.yur@gmail.com
Received: 11 July 2021; Accepted: 17 May 2021
Cryopreservation of spermatozoa is widely used in the treatment of infertility by assisted reproductive technologies. However, the cryopreservation causes an oxidative stress which can induce pathological changes in the male gametes. The aim of the research was to evaluate lipid peroxidation (LPO) and DNA fragmentation as well as correlation between these parameters in the fresh and cryopreserved spermatozoa of men with normozoospermia or oligoasthenoteratozoospermia (OAT) and spermatozoa derived from the epididymis of men with azoospermia. The level of malondialdehyde (MDA) in a TBA test, superoxide dismutase (SOD) activity and total antioxidant activity (AOA) were assessed. DNA integrity was estimated by acridine orange staining technique. It was shown that MDA level and SOD activity were significantly higher in the fresh spermatozoa of oligoasthenoteratozoospermic group compared with the fresh spermatozoa of normozoospermic group. After cryopreservation the MDA level increased in all groups and was the highest in OAT group where the greatest AOA decline was detected. DNA fragmentation frequency was 2.6 and 4.1 times higher in the fresh and cryopreserved OAT spermatozoa respectively as compared with the fresh normozoospermic ones (7.2%). DNA fragmentation was found to be the lowest in the fresh (6.2%) and cryopreserved (5.8%) epididymal spermatozoa. After cryopreservation SOD activity in epididymal spermatozoa was lower than in normozoaspermic. In spermatozoa of the studied groups the MDA level positively correlated with DNA fragmentation (0.79 Pearson’s correlation coefficient) both before and after cryopreservation. It is suggested that due to the detected low DNA fragmentation and LPO level in epididymal spermatozoa their use could be an alternative approach for infertility treatment by assisted reproductive technologies.
Astrocyte specific proteins content in the different parts of the rat and mongolian gerbil brain during ontogenesis
Y. P. Kovalchuk*, H. N. Shiyntum, G. A. Ushakovа
Oles Honchar Dniprо National University, Ukraine;
*e-mail: yulka.5868152@ukr.net
Received: 28 October 2021; Accepted: 17 May 2021
Astrocyte-specific proteins are used as markers of astrocytes, particularly in the case of age-related brain dysfunctions and neurodegenerative disorders. However, the data on the content of these proteins in different parts of the brain during ontogenesis are insufficient. In this research, the content of astrocyte specific Ca2+-binding protein S100B and glial fibrillary acidic protein (GFAP) in cerebellum, thalamus, and hippocampus of Wistar rats and Mongolian gerbils during ontogenesis were determined. Animals were divided by age into four groups (n = 6-10): 1 – newborn (1-day-old); 2 – 30-day-old; 3 – 90-day-old; 4 – 180-day-old. Fractions of soluble and filamentous proteins from the different parts of the brain were obtained by differential centrifugation and solubilization with 4M urea. The quantitative levels of S100B and GFAP were determined with ELISA. It was revealed that the content of S100B protein increased significantly by day 180 in all studied parts of the rat and gerbil brain. The content of GFAP both soluble and filamentous forms in the brain of 1 day-old animals was low, but raised significantly after 30 days in all studied parts of gerbil brain, and increased gradually up to 180 days in the rat brain. The magnitude of the increase in the content of the studied astrocyte-specific proteins was shown to be different and to depend on the brain part, postnatal development stage of the animals and their species.
Regulation of ionized calcium concentration in mitochondria matrix in the absence of exogenous Са(2+)
A. V. Sylenko*, S. G. Shlykov, L. G. Babich,
О. Yu. Chunikhin, S. O. Kosterin
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: sylenkoanna@ukr.net
Received: 15 September 2021; Accepted: 17 May 2021
Functional activity of mitochondria depends on ionized calcium content in its matrix. Thus, investigation of the ways to influence Ca2+ transport and accumulation in these organelles is an urgent issue. The aim of this work was to evaluate ionized calcium concentration in the mitochondria matrix in the absence of exogenous Ca2+ and in the presence of Mg2+ and ATP in incubation media. The concentration of ionized calcium in mitochondria isolated from the myometrium of non-pregnant rats was measured with the fluorescent probe Fluo-4 AM, mitochondria hydrodynamic diameter was assessed using dynamic light scattering. The ATP concentration-dependent increase in ionized calcium concentration in mitochondria matrix in the absence of exogenous Са2+ and the high level of the total Са2+ accumulation after further addition of exogenous Са2+ were registered. The effect of ATP on Са2+ concentration did not depend on the blockage of mitochondrial PTP with cyclosporine A or on H+-ATPase/ATP-synthase inhibition with oligomycin, instead the addition of 10mM theophylline and 30 mM NaHCO3 to the Mg2+-containing medium led to Са2+ concentration elevation in mitochondria matrix. It was shown that low Ca2+ concentration in the matrix correlated with the increase in mitochondria size, whereas high Ca2+ concentration correlated with relatively smaller size of the organelles. Activation of the soluble adenylyl cyclase by NaHCO3 with simultaneous inhibition of phosphodiesterase by theophylline was accompanied by the decline in Са2+ probe normalized fluorescence in mitochondria matrix under conditions of exogenous Ca2+ introduction. It was suggested that the soluble adenylyl cyclase may be involved in regulation of Са2+ concentration in mitochondria matrix.