Резюме
Корнієнко В.І.¹, Самура Б.А.², Романенко Н.І.³ Вплив бенфураму на ліпідний обмін при експериментальній гіперхолестеринемії.
Експериментальна гіперхолестеринемія у тварин  супроводжувалася зниженням рівня ліпопротеїдів високої  щільності (ХС-ЛПВЩ) на 22,2% з одночасним  збільшенням вмісту загального  холестерину (ЗХС) в 2,11 рази, підвищенням рівня ліпопротеїдів низької щільності (ХС-ЛПНЩ) в 5,89 рази, ліпопротеїдів дуже низької щільності  (ХС-ЛПДНЩ) в 2,59 рази,  загальних ліпідів в 1,61 раза и триацилгліцеролів в 2,52 рази. Проведене лікування експериментальної  гіперхолестеринемії  бенфурамом сприяло підвищенню рівня  ліпопротеїдів високої  щільності (ХС-ЛПВЩ) на 18,1%  з одночасним  зниженням в плазмі крові щурів вмісту загального холестерину на 37,8%, зменшенням  вмісту  ХС-ЛПНЩ на 22,8%, ліпопротеїдів дуже низької щільності  (ХС-ЛПДНЩ) – на 68,6%, триацилгліцеролів на 59,1% та загальних ліпідів на 3,5%  у порівнянні з ЕГ. Бенфурам є перспективною сполукою для подальшого  вивчення специфічної активності з метою створення  ефективного препарату для регуляції ліпідного обміну.
Ключові слова: бемфурам, експериментальна  гіперхолестеринемія, ліпідний обмін, нікотинова кислота.
Резюме
Корниенко В.И, Самура Б.А., РоманенкоН.И. Влияние бенфурама на липидный обмен при экспериментальной гиперхолестеринемии.
Экспериментальная гиперхолестеринемия у животных спровождалась снижением уровня липопротеидов высокой плотности (ХС-ЛПВП) на 22,2% с одновременным увеличением содержания общего холестерина (ОХС) в 2,11 раза, повышением уровня липопротеидов низкой плотности (ХС-ЛПНП) в 5,89 раза, липопротеидов очень низкой плотности (ХС-ЛПОНП) в 2,59 раза, общих липидов в 1,61 раза и триацилглицеролов в 2,52 раза. Проведенное лечение экспериментальной гиперхолестеринемии бенфурамом способствовало повышению уровня липопротеидов высокой плотности (ХС-ЛПВП) на 18,1%  с одновременным снижением в плазме крови крыс содержания общего холестерина на 37,8%, уменьшением содержания ХС-ЛПНП на 22,8%, липопротеидов очень низкой плотности (ХС-ЛПОНП) – на 68,6%, триацилглицеролов на 59,1% и общих липидов на 3,5% по сравнению с ЭГ. Бенфурам является перспективным веществом для дальнейшего изучения специфической активности с целю создания эффективного препарата для регуляции липидного обмена.
Ключевые слова: бемфурам, экспериментальная гиперхолестеринемия, липидный обмен, никотиновая кислота.
Summary
Kornienko V.I., Samura B.A., Romanenko N.I. Benfuram’s effect on lipid metabolism under experimental hypercholesterolemia conditions.
Experimental hypercholesterolemia in animals was accompanied by the reduce of the high density lipoproteins (HDL-C) level by 22.2% with a simultaneous increase in total cholesterol (TC) of 2.11 times, increased of low density lipoprotein (LDL-C) levels in 5.89 times and very low density lipoproteins (LDL-VLDL) by 2.59 times, the common lipid in 1.61 times and in 2.52 times for triacylglycerols. The treatment of the experimental hypercholesterolemia by benfuram helped to raise the high density lipoprotein (HDL-C) level by 18.1% with a simultaneous decrease of total cholesterol by 37.8% in rat plasma, decrease in the LDL-C amount by 22.8 %, very low density lipoproteins (VLDL-cholesterol) - 68.6%, triacylglycerol 59.1%, total lipids for 3.5% in comparison with EH. Benfuram is a promising substance for further search of the specific activity with the purpose to create an effective drug for the lipid metabolism regulation.
Key words: bemfuram, experimental hypercholesterolemia, lipid metabolism, nicotinic acid.
Рецензент: д.вет.н., проф. І.О. Жукова

УДК 615.011:547.857.4

¹Харківська державна зооветеринарна академія

Харьковская государственная зооветеринарная академия

Украина, 62341, Харьковская обл., Дергачевский район,пгт. Малая Даниловка

Kharkiv state zooveterinary academy

62341, Kharkivdistrict, Dergachivskiy region, Malaya Danilivka

²Національний фармацевтичний університет (Харків)

Национальный фармацевтический университет (Харьков)

61002, г. Харьков, ул. Пушкинская, 53

National University of Pharmacy (Kharkov)

Ukraine, 61002, Kharkov, 53, str. Pushkinska

³Запорізький державний медичний університет

Запорожский государственный медицинский университет

пр. Маяковского, 26, 69035, г. Запорожье, Украина

Zaporozhye State Medical University

69035, Mayakovsky avenue 26, Zaporozhye, Ukraine

kornienko-valentina1966@mail.ru

Introduction. Lipid metabolism disorders occupy one of the core places in the developed countries. Lipids are contained in the blood and deposited in the tissues. They are an essential component of cells and contribute to the normal function of the body. Disorders of fat metabolism, high cholesterol level lead to atherosclerotic disease of the coronary arteries, cause heart attack or ischemic stroke [4, 13, 17].

In a group of diseases with lipid disorders main focus is on the hypercholesterolemia, since between high levels of cholesterol - low-density lipoprotein and high-risk of coronary heart disease (CHD) there is a causal relationship. Currently, the concentration of total cholesterol in the blood to the risk of mortality from major diseases associated with hypercholesterolemia (CHD, cerebral infarction) is significant [8, 14].

Reducing the concentration of total cholesterol in the blood and increasing of the – HDL-cholesterol reduces the pathological conditions propagation [11]. Reduction of blood cholesterol by 10% leads to a reduction of heart disease mortality by 20%. Increasing the HDL-cholesterol concentration by 0.03 mmol/l reduces the risk of coronary heart disease by 2-3%. Regardless of the total blood cholesterol the relationship between the HDL-cholesterol contain and the heart disease frequency is stored [4, 12].

Increasing of LDL-cholesterol above 2.59 mmol/l is observed while rich in animal fat and cholesterol food consumption. Hypodynamia, unbalanced diet contribute to the development of overweight, hypertension, and diabetes are factors that increase risk of coronary heart disease [13, 16].

The concentrations of HDL-cholesterol less than 1.3 mmol/l and triglycerides in the blood of more than 2.3 mmol/l are predictive indicators of the probability of death from coronary heart disease. Therefore, the HDL-cholesterol level less than 1.3 mmol/l is assumed as the more accurate predictor of mortality against CHD than the concentration of total cholesterol [10, 14].

Reducing the LDL-cholesterol level - is a part of the program to reduce the risk of cardiovascular disease [1, 8]. Smoking cessation leads to an increase in HDL-cholesterol level. Similar properties have LDL-cholesterol, overweight and physical-load [5, 13, 15].

Drinking of 80 ml per day increases the amount of cholesterol and reduces cases of coronary heart disease. A doctor may not recommend alcohol drinks to a patient, but do not prevent the initiative of the patient, if their needs are included in the given scope. [8]

Effect of lipid-lowering agents should be aimed on reducing of LDL, VLDL levels and increase of HDL levels. According to these parameters statins are leading compounds, which are effective in conditions of hypercholesterolemia, secondary hyperlipidemia, developing while poor nutrition, renal and endocrine diseases, while steroids, estrogens, thiazide diuretics therapy [7, 12]. Hyperlipidemiс drugs are contraindicated while hepatic disfunction, renal failure, pregnancy, reducing their therapeutic value. In connection with this research for more effective and safe drugs for the correction of a lipid metabolism is an actual problem of experimental pharmacology.

Connection with academic scientific programs, plans, themes. The research was performed as a part of scientific and research work of the Kharkiv State Academy of Animal Health, National University of Pharmacy and Zaporozhye State Medical University on the topic "Obtaining, physical and chemical properties, biological activity and the study of xenobiotics effect on biological processes" (state registration №0105U002815, code IN 15.00.02.01).

The aim of this research was to investigate the effect of benfuram on lipid metabolism while experimental hypercholesterolemia.

Materials and methods.

The study was performed on Wistar rats weighing 185-215 g. Experiments with animals were performed in compliance with the principles of humanity, according to directives of the European Community (86/609/EES), and of the declaration of Helsinki of the World Medical Association on humane treatment with animals (1996). The experimental rats were divided into 3 groups of 6 animals each: 1st - intact, 2nd – with the experimental hypercholesterolemia, 3rd - with experimental hypercholesterolaemia with benfuram administration (intragastrically in daily dose of 50 mg/kg at 9:00 am, feeding time at 12 hours, that decreased possible benfuram’s interaction with food components), 4th group – with experimental hypercholesterolaemia with administration of reference drug - nicotinic acid at a dose of 300 mg/kg.

Experimental hypercholesterolaemia was simulated by adding of cholesterol diet (4%), 6 methyluracil (12%), an oil sunflower (25%) for 14 days [2]. For animal blood animals were decapitated under ethyl ether narcosis. Blood was collected in a clean tube with heparin solution, settled for 10 min and centrifuged at 3000 rpm for 10 min. Before the determination, plasma was stored in a freezer at -20 °C. Lipid profile of blood serum was tested by biochemical «Lasistems» analyzer “FP-901” (Finland). Condition of the lipid metabolism was assessed on the content of serum total lipids, triacylglycerols (TG), total cholesterol and HD-lipoproteins and LD- lipoproteins. The content of total lipids (TL) in the serum of rats was determined by a method based on the color reaction of decomposition products of unsaturated lipids with sulfophosphovaniline reagent: triacylglycerol (neutral fats) – by colorimetric method [3]; total cholesterol (TC) by the Ilk, HD-lipoprotein cholesterol β-(HDL-C) - by the method based on the precipitation of β-lipoproteins with heparin in the presence of magnesium ions in the blood serum of rats. The level of cholesterol of very low density lipoproteins (LDL, VLDL) was calculated as TG/2.2 mmol/l. The content of low density lipoprotein cholesterol was calculated using W.T.Fridewald equation: LDL-C = TC-(HDL-C) - (TG/2,2) and atherogenic index (AI) was calculated as follows: AI = TC-(HDL: VHDL) [16].

The experimental data were processed by conventional statistical methods using the Microsoft Office Excel 2003 software. The significance of differences between the experimental groups were evaluated using the Student t-test and a “STATISTICA” computer program for Windows 6.0» [6, 9].

Results and discussion.

Results of benfuram’s effect research on animals with experimental hypercholesterolaemia lipid metabolism are shown in Table 1. The data indicate that in laboratory rats in group 2, which were on 14 daily atherogenic diet an experimental alimentary hypercholesterolemia was formed. In the blood serum of rats in this group a reduction in HDL-C level by 22.2% and rising of LDL-C in 5.89 times were observed. Rising of LDL-cholesterol levels can stimulate the deposition of the resulting higher than normal cholesterol on the walls of blood vessels. The rapid rising of the LDL-C level may indicate a negative impact of EH on lipid metabolism. It can be assumed that storing on vessel walls, LDL particles lead to the development of atherosclerosis and its complications - myocardial infarction or ischemic stroke. Direct correlation between raised LDL-cholesterol levels and coronary heart disease [1] has been established.

The animals in group 2 with EH show raised total cholesterol (TC) level in 2.11 times and increased very low-density lipoprotein (VLDL) level in 2.59 times, an increase in total lipids in 1.61 times triacylglycerols – in 2.52 times. Direct correlation between increased levels of LDL-cholesterol and the incidence of coronary heart disease has been shown. A course of treatment by benfuram helped raise high density (HDL-C) by 18.1%, with a simultaneous decrease in total cholesterol by 37.8%, decrease of the LDL-C by 22.8%, very low-density (VLDL) - by 68.6%, 59.1% triacylglycerols and total lipids by 7.4% compared to the animals in group 2 with EH. Reducing the concentration of LDL-cholesterol and raising of  HDL-cholesterol concentrations suggests therapeutic effect of benfuram in experimental hypercholesterolemia of laboratory animals [12]. It can be assumed that the mechanism of benfuram’s action on lipid metabolism may be associated with an effect on low-density lipoproteid receptors [4].

Nicotinic acid in a dose of 300 mg/kg after intragastric administration caused a decrease in total cholesterol (TC) in 1.61 times and increase in high density lipoprotein (HDL-C) level by 9.4% and decreased the very low-density lipoproteins (VLDL) by 1.89 times and triacylglycerols – in 1.79 times and total lipids by 6.3%.

Thus, our studies have shown that in experimental hypercholesterolemia under the influence of benfuram and reference drug nicotinic acid the shift of the majority of the lipid metabolism indices in the blood serum were observed to be aside normalization.

Conclusions

1. While experimental hypercholesterolemia under the influence of benfuram and nicotinic acid the shift of the majority lipid metabolism indices aside normalization were observed.

2. Benfuram is a promising organic compound for subsequent research of the pharmacological properties and safety in order to create an effective drug for the correction of lipid metabolism on its basis.

Литература

1. Возиян П.А. Структурная организация липопротеинов плазмы крови и их взаимодействие с клеткой / П.А. Возиян, Ю.Д. Холодова // Укр. Биохим.журн. - 1989. – Т. 61, № 3. – С. 22-37.
2. Доклінічні дослідження лікарських засобів / За ред. О.В. Стефанова. – К.: Авіцена, 2001. – С. 433-443.
3. Клинико-биохимическая лабораторная диагнотика: справочник в 2-х томах / Под ред. В.С. Камышникова. – [2-е изд.]. - Минск: Интерпрессервис, 2003. – С. 106-177.
4. Климов А.Н. Обмен липидов, липопротеидов и его нарушения / А.Н. Климов, Н.Г.Никульчева. - СПб.:Питер Ком, 1999. – 512 с.
5. Компендиум-2007 – Лекарственные препараты / Под ред. В.Н. Коваленко, А.П.Викторова. – К.: Морион, 2007. – 2270 с.
6. Лапач С.Н. Статистические методы в медико-биологических исследованиях с использованием EXCEL / С.Н. Лапач, А.В. Чубенко, П.Н.Бабич. – К.: Морион, 2000. – 320 с.
7. Машковский М.Д. Лекарственные средства / М.Д. Машковский. – [15-е изд., перераб., испр. и доп.]. – М.: Новая волна, 2009. – 1206 с.
8. Сернов Л.Н. Элементы экспериментальной фармакологии / Л.Н.Сернов, В.В. Гацура. – М.: Медицина, 2000. – С. 308-328.
9.  Смирнов А.В. Характеристика дислипопротеидемии у больных гломерулонефритом / А.В. Смирнов // Нефрология. – 1998. – Т.2, № 3. – С. 76-83.
10.  Титов В.Н. Атеросклероз как патологияполиеновых жирных кислот. Биологические основы теории атерогенеза / В.Н. Титов. - М.: Фонд «Клиника ХХI века», 2002. – 495 с.
11.  Титов В.Н. Фундаментальная медицина. Различие переноса жирных кислот в липопротеинах очень низкой плотности и низкой плотности и биологические основы коррекции семейных гиперлипопротеинемий / В.Н. Титов // Клиническая лабораторная диагностика. –2005. – № 7. – С. 3-10.
12. Effects of loop diuretics on angiotensin II-stimulated vascular smooth muscle cell growth / P. Muniz, A. Fortuno, G. Zalba [е.а.] // Nephrol. Dial. Transplant. – 2001. – Vol. 16, № 1. – P. 14-17.
13. Сastello I.B. Hyperlipidemia. A risk factor for chronic allograft dysfunction / I.B. Сastello // Kidney Inf. - 2002. – Vol. 61, № 63. – P. 1964-1976.
14.  Castrop H. Modulation of adenosine receptor expression in the proximal tubule: a novel adaptive mechanism to regulate renal salt and water metabolism / H. Castrop // Am. J. Physiol. Renal. Physiol. – 2008. – Vol. 295, № 1. – P. 35-36.
15. Ross R. Atherosclerosis: an inflammatory disease / R.Ross // New Engl. J. Med. - 1999. - Vol. 340.-. P. 115-126.
16. Ross R. Atherosclerosis: a question of endothelial integrity and growth cjntrol of smooth muscle / R.Ross // The Harvey Ltctures. - 1983. – Ser. 77, № 4. – Р. 161-181.