BioDiscovery : Research Article
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Corresponding author: Vladimir Kulchitsky (vladi@fizio.bas-net.by)
Academic editor: Nikolai Zhelev
Received: 30 Jun 2017 | Accepted: 08 Aug 2017 | Published: 18 Oct 2017
© 2017 Khalil L. Gainutdinov, Svetlana G. Pashkevich, Vyatcheslav V. Andrianov, Guzel G. Yafarova, Margarita O. Dosina, Tatiana Kh. Bogodvid, Julia P. Stukach, Dinara I. Silant'eva, Aleksandra S. Zamaro , Timur V. Sushko, Vladimir Kulchitsky
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation: Gainutdinov K, Pashkevich S, Andrianov V, Yafarova G, Dosina M, Bogodvid T, Stukach J, Silant'eva D, Zamaro A, Sushko T, Kulchitsky V (2017) Participation of NO-synthase in Control of Nitric Oxide Level in Rat Hippocampus after Modelling of Ischaemic and Haemorrhagic Insult. BioDiscovery 20: e14810. https://doi.org/10.3897/biodiscovery.20.e14810
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Electron paramagnetic resonance (EPR) was used as a method for recording the content of the nitric oxide (NO) in hippocampal tissues of intact rats and rats after modelling of ischaemic and haemorrhagic stroke. Based on direct measurements of NO by EPR spectroscopy, it was shown that, within 5 hours after the onset of symptoms of ischaemic and haemorrhagic stroke, the formation of NO in the hippocampus was reduced by a factor of 2-3 and this reduction was maintained for a period of between 24 and 72 hours. The results show that a systemic character of a decrease in the intensity of NO production during the modelling of ischaemic events in the brain reflects the effects of central dysregulation of the functions at the level of the whole organism such that it is appropriate to consider implementing the correction of the vital systems of the body in a stroke. It has indicated that non-selective NO-synthase blocker L-NAME reduced the low level of NO production by a factor of 3 by its administration within 72 hours after post-ischaemic and haemorrhagic stroke. It was discovered however that L-NAME returns the level of NO production to baseline (control) by its administration within 5 hours after ischaemia.
Nitric oxide, Electron paramagnetic resonance, Spin trap, Ischaemic brain stroke, Haemorrhagic brain stroke
There appears to be increasing evidence about the impact of regulatory substances of intestinal microflora on the functional state of the brain of animals (
In the vital functions of animals, the role of NO is especially significant in the cardiovascular function (
Modelling of ischaemic and haemorrhagic stroke was produced on rats in the Institute of Physiology of NAS of Belarus, Minsk. Animals were kept in standard vivarium conditions (12/12- light and dark rhythm, air temperature of 23±1°C and stable supply and exhaust ventilation) with free access to water and food (ad libitum) and a diet in accordance with the standards for keeping laboratory animals. For the modelling of ischaemic stroke, animals were subjected to 5-minute hypoxia (conditional rise to a height of 4500m above sea level which corresponded to a pressure of 432mmHg and a decrease in oxygen partial pressure pO2 from 159mmHg to 90mmHg on average) (
The difficulty in determining the maintenance of the free NO in the tissues of the organism is due to its short lifetime which appears in low concentrations in tissues. Recently, one of the most effective methods for the detection and quantification of NO in biological tissues is the method of electron paramagnetic resonance (
The results are shown as mean ± SEM. The unpaired Student’s t-test and non-parametric Mann–Whitney test were used for comparison between two groups. One-Way ANOVA followed by the Tukey post-hoc test and a repeated Two-Way ANOVA were used for comparison between statistical groups. The statistical software SigmaStat32 was used. The statistical significance criterion was p<0.05.
Fig.
Fig.
The relative content of NO in the hippocampus of healthy rats (Control) and rats after 5 (Ischaemia, 5h) and 72 (Ischaemia, 72h) hours of the haemorrhagic stroke and also after using of inhibitor L-NAME (Ischaemia, 5h + L-NAME) and (Ischaemia, 72h + L-NAME) respectively. The ordinates axis is the average integral intensity of the signal.
Fig.
The relative content of NO in the hippocampus of healthy rats (Control) and rats after 5 (Ischaemia, 5h) and 72 (Ischaemia, 72h) hours of the ischaemic stroke and also after using of inhibitor L-NAME (Ischaemia, 5h + L-NAME) and (Ischaemia, 72h + L-NAME) respectively. The ordinates axis is the average integral intensity of the signal.
Taking into account that the NO may play a pathogenic role in a number of pathological conditions of the nervous system, including ischaemia, some NOS inhibitors have become the subject of intensive study as potential neuroprotective agents (
The problem of cerebral ischaemia is most acute in the world (
At the present time, the development of cerebral ischaemia and the subsequent occurrence of stroke are associated with impaired cerebral blood flow, as well as with impaired regulation of the blood supply to the brain tissues by the system of NO (
Thus, the analysis of literature and the results of our experiments show the ambiguity of the data that reflects the well-known fact of dose-dependent effects of NO in the brain. In the present work, it was demonstrated that the process of the comprehensive approach and the application of precision methods for measuring NO levels made it possible to receive data for the dynamics of NO production in nerve tissue.
The work is performed according to the Russian Government Program of Competitive Growth of Kazan Federal University; by Russian Foundation for Basic Research (Grant No. 16-54-00098) and by Belarusian Republican Foundation for Fundamental Research (Grant B16R-166). The authors thank V.S. Iyudin for help in EPR measurements.
Khalil L. Gainutdinov carried out research using electron paramagnetic resonance and formalised the results of this section of the study. He formulated the conclusions of the work. Svetlana G. Pashkevich conducted experimental studies in rats and formalised the results of the study. Vyatcheslav V. Andrianov carried out research using electron paramagnetic resonance and formalised the results of this section of the study. Guzel G. Yafarova carried out research using electron paramagnetic resonance and formalised the results of this section of the study. Margarita O. Dosina conducted experimental studies in rats and formalised the results of the study. Tatiana Kh. Bogodvid carried out research using electron paramagnetic resonance and formalised the results of this section of the study. Julia P. Stukach conducted experimental studies in rats and formalised the results of the study. Dinara I. Silant'eva carried out research using electron paramagnetic resonance and formalised the results of this section of the study. Aleksandra S. Zamaro conducted experimental studies in rats and formalised the results of the study. Timur V. Sushko conducted experimental studies in rats and formalised the results of the study. Vladimir Kulchitsky conducted experimental studies in rats and formalised the results of the study. He formulated the conclusions of the work.
No potential conflict of interest was disclosed by any of the authors.