The article is published in Ukrainian in the journal.
The English text is given in the author's version.
UDC 616.12-008.318-06:612.013.7
THE DYNAMICS OF THE ACTIVITY OF LIPID PEROXIDATION UNDER CONDITIONS OF ACUTE REGIONAL MYOCARDIAL ISCHEMIA.
KryghnaS.I., BereznyakovaM.E., Litvinova O.М., Fomina G.P., Karabut L.V.,AvidzbaYu. N.
National Pharmaceutical University, Kharkov City
Key words: hinolinamidetane, lipids, membranes, peroxidate oxidation.
The influence of hinolinamidetane on activity peroxidate oxidation of lipids isinvestigated at regional ischemia of the heart muscle. Is shown, that antioxidantical effect of hinolinamidetane is implemented at the expense of lowering a contents of primary molecular products of peroxidate oxidation of lipids in a tissues and blood.Results of biochemical research were confirmed by submicroscopic examination of myocardium, when attention was paid to ultrastructure of cardiomyocyte nucleus, sarcoplasm, availability of mitochondria in juxtanuclear area, borders of “insert disc” between sarcomeres, etc. After administration of quinoline-amide-ethane in animals, number of mitochondria significantly increased, borders of “insert discs” between sarcomeres slightly “expanded”, a lot of different organelles appeared in sarcoplasm. Restoration of myocardiocyte ultrastructure was confirmed by detected changes which appear as a result of hypoxia and in case of antioxidant action of both, tocopheryl acetate and quinoline-amide-ethane. Data сontent of conjugated dienes and trienes in myocardium and blood serum of animals show that vitamin E and quinoline-amide-ethane significantly inhibited generation of both, primary and secondary products of free-radical oxidation. Unidirectionality of shifts in myocardium and blood serum is also notable.Results of studying the content of Schiff bases showed that Vitamin E and quinoline-amide-ethane inhibited generation of such bases both, in myocardium and in blood serum. In comparison of agents’ activity, it was found that quinoline-amide-ethane to a greater degree prevented accumulation of peroxidation products.
Introduction
Interest in the use of quinoline-carboxylic acids and their derivatives for creation of pharmaceuticals is caused by the important part which their metabolites play in mitochondrial energy processes and the high efficacy which they demonstrate during various pathological conditions accompanied by disorders of energy metabolism and tissue respiration [5, 7].
Work objective is to study the impact of quinoline-amide-ethane substance on the activity of lipid peroxidation under conditions of acute regional myocardial ischemia.
Work connection with scientific programs, plans and topics. This research was performed in accordance with the plan of scientific and research works of the National Pharmaceutical University on the topic of “Pharmacological research of biologically active substances and pharmaceuticals of synthetic and biological origin, their application in medical practice”.
Materials and methods of research
Studies were conducted on 21 male rats weighing 180-200 g.Acute regional myocardial ischemia was caused by RB Jennings method under ethaminal-sodium anesthesia (40 mg/kg intravenous) [11]. Activity of lipid peroxidation and key anti-oxidase system enzymes was determined by means of sampling in decapitated animal blood serum and myocardium. The latter was homogenized in five-fold volume of molar saline solution, centrifuged under 41.67s-1 within 15 min., and supernatant was taken for examination. In serum and myocardium, content of TBA-products (conjugated dienes and trienes), Schiff bases (fluorescent products) and superoxide scavenger activity under method of V.B. Garvrilov and M.I. Mishkorudnaya, were studied [9, 10].
Degree of injury to myocardium and its membranes was studied with electron microscope EM 100 BR by means of examining contrast uranyl acetates and citrates of lead of ultrathin myocardial sections.
All animals were divided into three groups: 1st – control group (biological control) without impact of pharmaceuticals, 2nd – animals receiving vitamin E, 3rd – rats receiving quinoline-amide-ethane intragastrically.
Quinoline-amide-ethane, which is a new derivative of quinoline-carboxylic acids, was administered in the dosage of 40 mg/kg of animal mass orally once per 24 hours. Vitamin E (natural antioxidant) was prescribed according to the same scheme in the form of 10% oil solution in the dosage of 1 ml containing 100 mg of synthetic α-tocopheryl acetate [8].
Obtained results and their discussion
There was detected a direct correlation (r = +0.58) between the content of free-radical-oxidation products in myocardium and degree of myocardiocyte injury, and reverse correlation (r = -0.52) with the content of superoxide dismutase. Tables 1-4 demonstrate comparative antioxidant efficacy of products (in percentage as compared to control).
Table 1.
Content of TBA-reactive primary products in myocardium and blood serum of animals (X ± Sx), % to control, п=7
Group / Myocardium / Blood serum1
2
3 / 100.0 ± 5.0
37.6 ± 5.3
26.4 ± 4.1 / 100.0 ± 6.3
38.0 ± 4.2
28.2 ± 3.3
*р <0.001
Table 2.
Content of conjugated dienes and trienes in myocardium and blood serum of animals (X ± Sx), % to control, п=7
Group / Myocardium / Blood serumdienes / trienes / dienes / trienes
1
2
3 / 100.0±12.2
82.2±8.5
22.6±5.4** / 100.0±10.5 72.5±12.5* 14.3±3.2** / 100.0±9.9
86.5±10.3
39.2±6.1** / 100.0±11.5 73.2±9.0*
26.4±5.6**
*р <0.05 as compared to control;
** р <0.001.
Data of Tables 1 and 2 show that vitamin E and quinoline-amide-ethane significantly inhibited generation of both, primary and secondary products of free-radical oxidation. Unidirectionality of shifts in myocardium and blood serum is also notable.
Results of studying the content of Schiff bases showed that Vitamin E and quinoline-amide-ethane inhibited generation of such bases both, in myocardium and in blood serum (Table 3). In comparison of agents’ activity, it was found that quinoline-amide-ethane to a greater degree prevented accumulation of peroxidation products.
Table 3.
Content of Schiff bases in myocardium and blood serum of animals (X ± Sx), % to control, п=7
Group / Myocardium / Blood serum1
2
3 / 100.0 ± 5.4
83.6 ± 4.5 *
52.6 ± 7.9 ** / 100.0 ± 5.1
91.0 ± 2.7 *
58.2 ± 5.1 **
* р <0.05 as compared to control;
** р <0.001.
Superoxide dismutase plays one of the important parts in tissue protection from free-radical oxidation [10]. It has been established that preliminary administration of the latter prevents from stress-ischemic and reperfusion injury of myocardiocytes, while preserving strength and rate of myocardium contraction, eliminates inhibiting impact of low pH on the rate of calcium absorption by sarcoplasmic reticulum. It is very important that superoxide dismutase detoxicates hydroxyl radicals from all sources of their production [9].
Increase of myocardium superoxide-dismutase activity and blood serum superoxide-eliminating activity has been detected under the impact of quinoline-amide-ethane and tocopherol (Table 4) [9, 10].
Conducted research showed that use of quinoline-amide-ethane significantly reduced the content of primary molecular products of lipid peroxidation in cardiac tissue and blood, reduced cholesterol/phospholipids ratio [2, 6], and stimulated superoxide-dismutase activity of myocardial tissue and superoxide-entraining activity of blood serum.
By its impact on lipid peroxidation processes, quinoline-amide-ethane had greater impact than vitamin E.
Results of biochemical research were confirmed by submicroscopic examination of myocardium, when attention was paid to ultrastructure of cardiomyocyte nucleus, sarcoplasm, availability of mitochondria in juxtanuclear area, borders of “insert disc” between sarcomeres, etc. After administration of quinoline-amide-ethane in animals, number of mitochondria significantly increased, borders of “insert discs” between sarcomeres slightly “expanded”, a lot of different organelles appeared in sarcoplasm [12]. Restoration of myocardiocyte ultrastructure was confirmed by detected changes which appear as a result of hypoxia and in case of antioxidant action of both, tocopheryl acetate and quinoline-amide-ethane.
Table 4.
Dynamics of growth of superoxide-dismutase activity of myocardium and blood serum (X ± Sx), % to control, п=7
Group / Myocardium / Blood serum1
2
3 / 100.0 ± 5.4
128.8 ± 7.5 *
220.1 ± 9.3 ** / 100.0 ± 6.1
118.3 ± 7.9 *
195.2 ± 10D **
* р <0.05 as compared to control;
** р <0.001.
Analysis of data in literature [1, 2, 3], which speak of the ability of tocopheryl acetate to improve metabolism and contractive activity of myocardium, reduce oxygen consumption by myocardium, take part in tissue respiration and in other important processes of cell metabolism, allows with great probability, while taking into account results of own research, assuming that quinoline-amide-ethane possesses similar properties.
Conclusion
1. Antioxidant effect of quinoline-amide-ethane is realized as a result of reduction of primary molecular products of lipid peroxidation in myocardium and blood, stimulation of superoxide-dismutase activity of cardiac muscle and superoxide-entraining activity of blood serum.
References
- Березнякова М.Є. Стан кардіогемодинаміки щурів при гострому інфаркті міокарда під впливом сучасних антигіпоксантів // Пробл. екол. та мед. –Полтавська укр. стомат. академія. - №3-4,2006. - С. 18-12.
- Храпак ВВ. Нормовані величини основних структурних елементів ЕКГ статевозрілих щурів-самців // В кн.: Доклінічні дослідження лікарських засобів (методичні рекомендації) / За ред. чл.-кор. О.В. Стефанова. - К.: Авіцена. 2007. - С. 271-283.
- Бугаенко В.В. Особенности внутрисердечной гемодинамики у пациентов и ишемической болезнью сердца со стабильной стенокардией и без стенокардии в зависимости от числа пораженных венечных артерии Укр. мед. часопис. - 2007. - № 6. - С. 94-98.
- Ватутин Н.Т., Калинкина Н.В., Долженко Е.В. и соавт. Безболевая ишемия миокарда // Укр. кардиолог, журн. - 2007. - № 3. - С. 97-103
- Воронков Л. Г. Есть ли перспективы у так называемой метаболической терапии при хронической коронарогенной дисфункции левого желудочка Взгляд сквозь призму доказательной медицины // Укр. кардиолог. журн. - 2009. - дод. 1. - С. 165-170.
- Литвицкий П.Ф. Патогенные и адаптивные изменения в сердце при его регионарной ишемии и последующем коронарного кровотока // Патол. физиол. и эксперим. терапия. - 2004. - №3. - С.2-11.
- Маколкин В. И. Ишемическая дисфункция миокарда и пути ее коррекции / Форум «Ишемическая болезнь сердца». - М., 2000. - №2. – С. 2-4.
- Сернов Л.Н. Гацура В.В. Элементы экспериментальной фармакологии - М., 2000 - 352 с.
- Сапрунова В.Б. Выявление цитохром С-оксидазной активности в митохондриях кардиомиоцитов изолированной ткани миокарда при длительном действии гипоксии / В.Б. Сапрунова, И.М. Солодовникова, Л.Е. Бакеева // Цитология – 2008. – Т.50 №3. – С. 268-274.
- Плужников Н.Н. Прооксидантная система. Подходы к выбору алгоритма исследования / Н.Н. Плужников // Весник оториноларингологии – 2010. – №5. – С. 72-75.
- Jennings R.B., Reimer К.А. Lethal myocardial ishemic injury // Am. J.Paton.- 1981. -Vol. 102. P.241-255.
- Haalliwel В. The biological effects of the superoxide radical and its products. Bull. Eur. Physio-path. Res. 1981; 21,17: 21-29.