ISSN 2415-3060 (print), ISSN 2522-4972 (online)
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УЖМБС 2017, 2(2): 26–30
Experimental Medicine

The Role of Disorders of Metabolism of Nitric Oxide for Pathogenesis of Experimental Bronchial Asthma

Kolishetska M. A., Regeda M. S.

The aim is to study the activity of nitric oxide synthase (endotelial and inducible), arginase activity of peripheral blood lymphocytes of Guinea pigs with experimental asthma. Materials and methods. All experiments on laboratory animals carried out with following the European Convention for the protection of vertebrate animals used for experimental and other scientific purposes (Strasbourg, 1986), Council Directive 2010/63 / EU, the Law of Ukraine 3447- IV “protection animals from the cruelty” the general ethics of animal experimentation adopted by the first national Congress on bioethics in Ukraine (2001). The researches were carried out on 72 Guinea-pigs (category: Cavia porcellus, males). The weight of each one was 180-220g. They were divided into 5 groups for 15 animals in each of them. Among the first group (control) there were intact Guinea-pigs. Among the second group there were animals with an experimental BA (on 5th day of experiment). Among the third group there were Guinea-pigs on the 19th day of the experiment. The fourth group consisted of animals with an experimental BA (on 26th day of the experiment) and the fifth group included Guinea-pigs with an experimental BA (33rd day). Two periods of experimental asthma were separated in order to make detailed analysis and interpretation of indicators of endogenous intoxication in different days of the experiment: early period and late one. The early period included a group of animals with BA on the 5th and 19th days of the experiment. Late period included Guinea pigs on 26th and 33rd day of asthma. The experimental asthma model reproduced in Guinea pigs by Babich method (1979). Animals were sensitized by a normal horse serum (0.1 ml intraperitoneally). Next three days they were entered subcutaneously 0.1 ml of normal horse serum (NKS) with a setting in an autoclave BCG (1 mg BCG 1.0 ml NKS). The next 14 days the animals were exposed to inhalation (using a spray) NKS to 1.0 ml of serum for each Guinea pig daily for 30 minutes in a sealed chamber. After this period every 7 days, Guinea pigs had these NKS inhalation. Then the animals were decapitated under the influence of ether anesthesia. In all groups of Guinea pigs there were determined NO synthase activity by specific cleavage NADPH (H +), arginase lymphocyte activity by the formation of urea. Numerical results were adapted with static method using Student’s criteria. Results of the research. The dynamics of formation of asthma found that arginase activity of blood lymphocytes increases almost linearly at an early period and reaches a maximum term experiment at the latest. Since the 5-day study of BA formation rate increased by 8.8% (r≤0,05) and after 19 days the activity of arginase is much progress relative to the control group (an increase of 46.8% (r≤0, 05)). In the later period of asthma continues increasing arginase activity, namely 65% (r≤0,05) and 87.8% (r≤0,05) respectively 26th and 33rd day modeling process when compared with intact animals. To assess the activity of endotelial NO-synthase (eNOS) it was recorded its reduction in the dynamics of the formation of experimental asthma. Thus, the level of eNOS on the 5th day was reduced by 12.4% (r≤0,05) compared with control values. Later, on the 19th day of the experiment it was observed gradual regression of the studied index by 23.9% (r≤ 0.05) compared with the first group of animals and 32.2% (r≤ 0.05) on the 26th day experimental model against the first group of animals. In the latest period on the 33rd day eNOS activity by 44.1% (r≤ 0.05) was lower compared to intact animals. Studying inducible No-synthase (iNOS) activity in peripheral blood lymphocytes of Guinea pigs in experimental asthma the following picture was observed: on 5th day iNOS was increased by 22.4% (p ≤ 0,05) compared with control values. Next, a 19 day process model was found and this figure has increased by 40.7% (p ≤ 0,05) the relative of groups of animals, and later, on the 26th and 33rd day of the experiment, it was found that it was even harder progression respectively 56.5% (p ≤0,05) and 93% (r≤0,05) compared with the control group. Conclusions. As a result of studies there were changes in the activity of arginase-NO-synthase system of blood lymphocytes, which improve arhinase activity of lymphocytes and increased activity of inducible NO-synthase and they were accompanied by compensatory inhibition activity of endothelial isoforms of NOS.

Keywords: bronchial asthma, nitric oxide, lymphocytes, arginase, NO-synthases

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