Tag Archives: nitric oxide

Nitric oxide cycle activity in rat biceps femoris muscle under conditions of bacterial lipopolysaccharide influence, experimental metabolic syndrome and their combination

O. Ye. Akimov1*, A. O. Mykytenko2, V. O. Kostenko1

1Department of Pathophysiology, Poltava State Medical University, Poltava, Ukraine;
*e-mail: o.akimov@pdmu.edu.ua;
2Department of Biological and Bioorganic Chemistry, Poltava State Medical University, Poltava, Ukraine

Received: 22 April 2023; Revised: 05 June 2023;
Accepted: 7 September 2023; Available on-line: 12 September 2023

There is evidence that long-term organism stimulation with bacterial lipopolysaccharides (LPS), which promotes the secretion of pro-inflammatory cytokines and nitric oxide, may play an important role in metabolic syndrome (MetS) development. Changes in NO production under conditions of MetS have different directions and depend on a specific organ. The purpose of this work was to study the production of nitric oxide and its metabolites in the biceps femoris muscle of rats under conditions of lipopolysaccharide stimulation of the organism, metabolic syndrome and their combination. The study was conducted for 60 days on 24 male Wistar rats divided into control, MetS, LPS and LPS+MetS groups. MetS was reproduced by adding 20% fructose solution to food, LPS stimulation was carried out by intraperitoneal injection of S. typhi LPS. It was demonstrated that stimulation of the rat organism with LPS under conditions of experimental metabolic syndrome increased the production of nitric oxide by L-arginine-dependent pathway, but limited metabolic syndrome-induced increase in nitric oxide production by L-arginine-independent pathway, reduced the concentration of S-nitrosothiols, while increasing the concentration of peroxynitrites and nitrites in the biceps femoris muscle of rats.

The level of nitric oxide and arginase activity in patients with arterial hypertension and diabetes mellitus during COVID-19

O. Y. Sklyarova1, S. R. Mahiiovych2, N. V. Denysenko3,
L. I. Kobylinska3*, Y. Y. Sklyarov2

1Department of Family Medicine FPGE, Danylo Halytsky Lviv National Medical University, Ukraine;
2Department of Therapy No 1 and Medical Diagnostics FPGE, Danylo Halytsky Lviv National Medical University, Ukraine;
3Department of Biological Chemistry, Danylo Halytsky Lviv National Medical University, Ukraine;
*e-mail: lesyaivanivna.biochemistry@gmail.com

Received: 28 September 2022; Revised: 06 November 2022;
Accepted: 11 November 2022; Available on-line: 19 December 2022

The aim of this study was to assess the level of nitric oxide production and arginase activity in patients with arterial hypertension and type II diabetes mellitus during infection with SARS-CoV-2. The study groups included patients with arterial hypertension, patients with arterial hypertension combined with a severe course of COVID-19 and patients who, in addition to arterial hypertension and COVID-19, were suffering from type II diabetes mellitus. The volunteers without any clinical signs of diseases and normal blood pressure formed the control group. It has been established that arterial hypertension, combined with COVID-19 occurs along with reduced L-arginine, nitric oxide, superoxide dismutase activity and increased arginase activity. At the same time, the presence of arterial hypertension in patients with diabetes and coronavirus disease is accompanied by a decline in the content of L-arginine and arginase activity. Our study’s results may help scientists find new pharmacological targets in the future treatment of coronavirus disease and comorbid disorders.

Involvement of Cu-containing amine oxidases in the development of lung pathology in ovalbumin-induced bronchial asthma in guinea pigs

O. Hudkova1*, S. Luhovskyi2, L. Drobot1, N. Latyshko1

1Palladin Institute of Biochemistry, National Academy of Sсiences of Ukraine, Kyiv;
2Dmitry F. Chebotarev Institute of Gerontology, National Academy of Medical Sciences of Ukraine, Kyiv;
*e-mail: ogudkova@biochem.kiev.ua

Received: 03 August 2022; Revised: 15 September 2022;
Accepted: 29 September 2022; Available on-line: 06 October 2022

Bronchial asthma is developed as an immune response to allergen challenges accompanied by inflammation and fibrosis implicated in airway remodeling. To reveal the causative implication of Cu-containing amine oxidases, semicarbazide-sensitive amine oxidase (SSAO), diamine oxidase (DAO), and lysyl oxidase (LOX), in bronchial asthma development we used their irreversible inhibitor, semicarbazide, and ovalbumin-induced pathology in guinea pigs. Semicarbazide was introduced to asthmatic animals via drink or inhalation. At the 16th week after disease induction, the increase in the activity of pro-inflammatory SSAO and DAO in plasma (1.6 and 2 times, respectively) was observed. The introduction of semicarbazide to asthmatic animals via drink or inhalation significantly decreased activities of these enzymes compared to the untreated asthmatic animals. A considerable­ increase in IL-13 content and LOX activity in the lung tissue of asthmatic animals were observed that evidenced airway inflammation and pulmonary fibrosis development. The uptake of semicarbazide by guinea pigs with bronchial asthma led to normalization of LOX activity. Histological studies confirmed that semicarbazide attenuated morphopathological changes in the lungs of asthmatic animals. Thus, the data obtained indicate the direct participation of the studied enzymes in the progression of pathological processes in atopic bronchial asthma as well as the potential use of semicarbazide as a drug in complex anti-asthmatic therapy.

Unraveling the mystery of nitric oxide: Nobel prize winners Robert Furchgott, Louis Ignarro, and Ferid Murad

T. V. Danylova1*, S. V. Komisarenko2

1Institute for Social and Political Psychology, National Academy of Educational Sciences of Ukraine, Kyiv;
*e-mail: danilova_tv@ukr.net,
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;

Received: 21 February 2022; Accepted: 01 July 2022

In the 21st century, none of the scientists denies the determining role of the cardiovascular system and its central organ, the heart. The ongoing attempts to design new medications, elaborate effective trainings, heart transplant programs testify that humanity does not abandon attempts to improve and prolong human life, especially given the fact that the world’s biggest killer is ischemic heart disease. The most significant achievements in this field receive the highest rating in the scientific community – the Nobel Prize. In 1998, the Nobel Prize in Physiology or Medicine was awarded jointly to Robert F. Furchgott, Louis J. Ignarro and Ferid Murad “for their discoveries concerning nitric oxide as a signaling molecule in cardiovascular system”. Their discovery triggered an international boom in research on nitric oxide. The paper aims to outline briefly the main stages of the scientific activity of R.F. Furchgott, L.J. Ignarro and F. Murad.

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.

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++ 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.

Indexes of nitric oxide system in experimental antiphospholipid syndrome

O. Z. Yaremchuk, K. A. Posokhova, І. P. Kuzmak,
M. I. Kulitska, I. М. Klishch, M. M. Korda

I. Horbachevsky Ternopil National Medical University, Ukraine;
e-mail: yaremchuk@tdmu.edu.ua

Received: 11 November 2019; Accepted: 21 January 2020

Antiphospholipid syndrome (APS) is an autoimmune disease characterized by the presence of antibo­dies to negatively charged membrane phospholipids (aPL).  Endothelial dysfunction is one of the most dangerous­ APS manifestations followed by thrombosis, placental insufficiency and often foetal death due to circulatory disorders in placenta blood vessels. It is established that synthesis and bioavailability of nitric oxide (NO) in the endothelium are impaired at APS, but the role of NO system in pregnancy failure at this pathology remains ambiguous. The aim of this research was to estimate the indexes of the nitric oxide system in animals with an experimental antiphospholipid syndrome before pregnancy and on the 18th day of pregnancy, without­ treatment and under treatment with nitric oxide synthesis modulators (L-arginine and aminoguanidine). In the blood serum and liver of the BALB/c mice with experimental APS, the content of eNOS and iNOS by ELISA and the level of NO2 and NO3 with the use of Gris reagent were determined before pregnancy and on the 18th day of pregnancy. The data obtained indicate the relative inefficient NO production by eNOS and NO hyperproduction by iNOS in the blood serum and liver of mice in the pathogenesis of experimental APS. Thus, in mice with APS before pregnancy and on the 18th day of the pregnancy, the eNOS content and NO2 level were decreased while the iNOS content and NO3 level were increased compared to the indexes in the control animal group. L-arginine administration to the animals with APS at the follow-up periods resulted in an increased eNOS content and NO2, NO3 levels in blood serum and liver with the simultaneous decrease in iNOS content in the liver as compared to indexes in untreated mice with APS. The combined use of L-arginine and selective iNOS inhibitor aminoguanidine caused a significant increase in eNOS content and a decrease in iNOS content followed by normalization of NO2 and NO3 levels in blood and liver of mice with  experimental APS before pregnancy and on the 18th day of pregnancy compared to untreated mice with APS.

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.

Effect of sodium nitroprusside and S-nitrosoglutathione on pigment content and antioxidant system of tocopherol-deficient plants of Arabidopsis thaliana

N. M. Semchuk, Yu. V. Vasylyk, O. I. Kubrak, V. I. Lushchak

Vassyl Stefanyk Precarpathian National University, Ivano-Frankivsk, Ukraine;
e-mail: lushchak@pu.if.ua

Sodium nitroprusside (SNP) and S-nitrosoglutathione (GSNO) were used as a source of exogenous nitric oxide (NO) to investigate their effects on biochemical parameters and antioxidant enzyme response in leaves of wild type Columbia and tocopherol-deficient vte4 and vte1 mutant lines of Arabidopsis thaliana plants and possible tocopherol involvement in regulation of antioxidant response under NO-induced stress. SNP enhanced the activity of the enzymes, that scavenge hydrogen peroxide in leaves of all studied lines, and increased glutathione reductase and glutathione-S-transferase activity there. In addition, it decreased the intensity of lipid peroxidation in vte1 mutant line leaves. At the same time, GSNO increased the levels of protein carbonyls and inactivated enzymes ascorbate peroxidase, guaiacol peroxidase and dehydroascorbate reductase in almost all investigated plant lines. In contrast to wild type, GSNO increased superoxide dismutase activity and decreased catalase activity and chlorophyll a/b ratio in the leaves of two mutant lines. It can be assumed that tocopherols in some way are responsible for plant protection against NO-induced stress. However the mechanisms of this protection remain unknown.