An effect of the indolinoren on the degree of hydration of the lung tissue in experimental pulmonary edema abstract

UDC 616.24-005.98:547.757
T. I. Tiupka, A. Yu. Markina, N. M. Kononenko, A. O. Minaieva
The National University of Pharmacy, Kharkiv
AN EFFECT OF THE INDOLINOREN ON THE DEGREE OF HYDRATION OF THE LUNG TISSUE IN EXPERIMENTAL PULMONARY EDEMA

ABSTRACT

In a study on rats found that the use of new derivatives of 2-oxindolin with diuretic properties – indolinoren – in experimental hemodynamic pulmonary edema reduces the degree of hydration of the lung tissue. Compared to hypothiazide, indolinoren has a pronounced anti-edema effect, as evidenced by a significant reduction in lung coefficient and increase the dry residue of lung tissue in experimental pulmonary edema.

Keywords: diuretics, pulmonary edema, pulmonary coefficient

INTRODUCTION
Pulmonary edema is a serious complication of various diseases associated with excessive impregnation of tissue fluid to the surface of the diffusion alveolar-capillary membrane [1]. The purpose of immediate therapeutic measures in pulmonary edema is a struggle with acute respiratory insufficiency and elimination of factors that affect the process of accumulation of water, electrolytes and proteins in the extravascular space of the lungs. Traditionally, patients with pulmonary edema is prescribed diuretics [6, 11]. They cause rapid effect because the effect on the venous circulation and reduce microvascular perfusion of the lungs. The principal mechanism of their pharmacological activity is the excretion by kidneys of sodium chloride and water. With the increase in urine output decreases final diastolic pressure in the cavity of the left ventricle, that, in turn, leads to a decrease in hydrostatic pressure in the pulmonary capillaries. These pharmacological properties of diuretics can reduce hydrostatic pressure, pulmonary microvascular filtration [8]. Of all the existing diuretic medicines preference is given to furosemide. Also used bumetanide, torsemide, ethacrynic acid, but with these drugs is associated a number of undesirable effects: with an increase in urine output increased loss of potassium and large amounts of chlorine, leading to cardiac arrhythmias [9, 10]. In this regard, the search for new diuretics to treat pulmonary edema is an actual problem of pharmacology.

The aim – to study the effect of new derivatives of acylated 2-oxindolin with diuretic properties (conventional name – "Iindolinoren") to the hydration degree of lung tissue in experimental hemodynamic pulmonary edema in rats.

MATERIALS AND METHODS

Hemodynamic pulmonary edema caused by the introduction of rats intramuscular epinephrine hydrochloride ("Zdorovya", Ukraine) at a dose of 5 mg/kg [4]. Experiments were conducted on 40 white male rats nonlinear weight 180-220 g, which were on a standard diet of food and water in accordance with sanitary regulations. During the experiment, animals treated in accordance with international principles of "European Convention for the protection of vertebrate animals used for experimental and other scientific purposes" (Strasbourg, 03/18/1986) and "General ethical animal research" (Ukraine, 2001). The experimental animals were divided into 4 groups: 1st group - intact control; 2nd group – rats with pulmonary edema without treatment; 3rd – rats with pulmonary edema, which was administered indolinoren; 4th – rats with pulmonary edema, which was injected drug comparison – hypothiazide. Indolinoren and hypothiazide administered 2 hours prior to simulation effective in swelling intragastric doses of 29.5 mg/kg and 25 mg/kg, respectively. Pre-anesthetized animals of the intact control deduced from the experiment by decapitation. Rats in the 2nd, 3rd and 4th groups died within 7-25 minutes of acute respiratory failure caused by adrenalin pulmonary edema.

The presence and degree of edema was assessed by lung coefficient size and the dry residue of lung tissue [7].

Pulmonary coefficient was calculated using the formula:

Also, the degree of hydration of the lung tissue was assessed by histological study. Pieces of lung tissue of all groups of animals were fixed in 10% formalin solution, dehydrated in alcohols of increasing strength, poured in celloidin-paraffin. Sections were stained with hematoxylin and eosin [3]. For ease of comparison, the state of the lung parenchyma of animals of different experimental groups were determined on specimens conditioned degree of severity of alveolar edema and preservation of alveolar pattern on a 5-point scale. The basis was semiquantitative visual assessment of capacity color of microstructures with histochemical reactions by the method of Sokolovsky [5]. Expression of alveolar edema was evaluated as follows: 0 points – no signs of edema; 1 point – little pronounced swelling (fluid found in the alveoli 1/3 of the drug); 2 points – moderately expressed swelling (fluid found in 1/3 of the alveoli in the microscope field of view); 3 points – pronounced swelling (fluid found in 2/3 of alveoli per field); 4 points – very distinct swelling (fluid found in almost 100% of the alveoli of the drug). The degree of preservation of alveolar pattern was evaluated according to the following scale: 0 points – histoarchitectonic of respiratory tissue department destroyed completely; 1 point – a typical pattern tissue saved to 1/3 square specimens; 2 points – 50% of the specimens; 3 points – at 2/3 square specimens; 4 points – a typical pattern of tissue can be traced throughout the area specimens.

Microscopy and photographed carried out the microscope micropreparations Micros 400 (Austria), beefed up with a digital camera Nikon Cool Pix 4500. Photographing held in the system Aver Media, pictures processed on a computer Pentium 2,4 GHz using Nikon View 5.

Results of the study were processed statistically parametric and non-parametric methods and nonparametric counterparts comparison of central tendencies in several groups (univariate analysis analogue – Kruskal-Wallis test, Mann-Whitney test, adjusted Bonferoni, dvovybirkovyy Kolmogorov-Smirnov test and the Wald-test series Volfovitsa). Data processing was carried out using the statistical software package Statistica 6.0 [2].

RESULTS AND DISCUSSION

Analysis of the results of the study showed that the use of indicated in the "Materials and Methods" ways of modeling pulmonary edema led to the development of its full clinical picture in all groups of experimental animals. On the severity of edema indicating a significant increase in lung weight coefficient and reduce the relative weight of the dry residue of tissue (Table 1).

Thus, in animals with pulmonary hemodynamic pulmonary edema factor increased almost 3.5 times as compared with intact control, where it stood at 0.65%. Since lung ratio may increase not only due to excess water in the lungs, but also as a result of increased blood supply, for a complete description of edema, we determined the dry residue of the lungs, which to a great extent reflects the degree of excess hydration. Found that the hemodynamic pulmonary edema dry residue of the lung tissue was in 1,5 times lower than in intact animals. The results are consistent with the literature, according to which the decrease in the dry residue of the lung tissue is likely a sign of accumulation of fluid and the degree of reduction depends on its severity of pulmonary edema [1].

In the 3rd and 4th groups of animals which were administered prophylactically indolinoren and hypothiazide, the severity of pulmonary edema was lower in comparison with the control diseases. This was evidenced in lung ratio, that decreased by 1.6 and 1 and 3 times, respectively. In addition, the application hypothiazide observed tendency to normalize the number of dry residue of the lungs, which increased in the 3rd and 4th group at 1.3 and 1.2 times, respectively.

Thus, prophylactic administration of indolinoren hypothiazide and animals with experimental pulmonary edema leads to a decrease in hydration effects in the lungs. The most pronounced anti-edema effect was noted in the application of the indolinoren.

To compare expression decongestants action during histological examination of lung tissue was performed semi-quantitative assessment of lung tissue of rats of different groups of the experiment (Table 2).

Analysis of the results of the study pointed out the effectiveness of indolinoren in experimental pulmonary edema. A statistically significant difference between efficiency and indolinoren drug comparison – hypothiazide not observed.

CONCLUSIONS

Prophylactic indolinoren in hemodynamic experimental edema reduces the degree of hydration in the pared to hypothiazide, indolinoren has a pronounced anti-edema effect, as evidenced by a significant reduction in lung coefficient and increase the dry residue of lung tissue in terms of hemodynamic edema in rats.

REFERENCES:

1. Гриппи легких / Гриппи; - М.: Бином, Х.: МТК-книга, 2005. – 304 с.

2. , , Бабич методы в медикобиологических исследованиях с использованием Ехсеl. – 2001. – 320 с.

3. Меркулов патологогистологической техники. – М.: Медицина, 19с.

4. , // Элементы экспериментальной фармакологии. - М.: Медицина, 2000. – С. 192.

5. Соколовский исследования в токсикологии. Л.: Медицина, 1971. – 176 c.

6. Сумин состояния /Сергей Александрович Сумин. – Омск: МИА, 2006. – 800 с.

7. І. Порушення водно-електролітного обміну при гемодинамічному і гострогіпоксичному набряку легень та можливі шляхи його корекції / Т. І. Тюпка// Запорожский медицинский журнал№3. - С. 36-39.

8. Шанин критических состояний /. –СПБ: Элби, 2003. – 435 с.

9. Штриголь ічні ефекти сечогінних засобів / , єва // Ліки№3-4. - С.58-64.

10. Groeneveld A. B. Vascular pharmacology of acute lung injury and acute respiratory distress syndrome / A. B.Groeneveld // J. Vascul. pharmacol. – 2002. – Vol. 39, № 4-5. – P. 247-256.

11. Mattu A. Cardiogenic Pulmonary Edema / A. Mattu // Emerg. Med. Clin. North. Am. – 2000. - Vol. 2. - P. 9-16.

Table 1.

Influence of the indolinoren on the degree of lungs hydration in hemodynamically pulmonary edema in rats, Х±Sx, n=10

Group number	Object of study	Pulmonary coefficient, %	The dry residue, %
1	Intact control	0,65±0,01	20,70±0,24
2	Hemodynamic edema	2,25±0,02*	13,97±0,09*
3	Hemodynamic edema + indolinoren	1,38±0,04*/**	18,24±0,22*/**
4	Hemodynamic edema + hypothiazide	1,75±0,02*	16,28±0,15*

Notes: p<0.05 * – relatively the intact control; ** – relatively animals with lung edema without treatment; *** – relatively reference drug hypothiazide.

Table 2.

Semi-quantitative assessment of lung tissue of rats, points

Group of experiment	Signs
Distinctiveness of alveolar edema (Ме[LQ; UQ])	Distinctiveness of alveolar pattern (Ме[LQ; UQ])
Intact control	0,0 (0,0; 0,0)	4,0 (4,0; 4,0)
Control Pathology	4,0 (3,0; 4,0)*	0,0 (0,0; 1,0)*
Indolinoren	2,0 (1,0; 2,0)*/**	2,0 (2,0; 3,0)*/**
Hypothiazide	1,0 (1,0; 2,0)*/**	2,0 (1,0; 2,0)*/**

Notes: p <0.05 * – relatively the intact control; ** – relatively the control pathology.

УДК 616.24-005.98:547.757

Т. И. Тюпка, А. Ю. Маркина, Н. Н. Кононенко, А.А. Минаева

ВЛИЯНИЕ ИНДОЛИНОРЕНА НА СТЕПЕНЬ ГИДРАТАЦИИ ЛЕГОЧНОЙ ТКАНИ ПРИ ЭКСПЕРИМЕНТАЛЬНОМ ОТЕКЕ ЛЕГКИХ

АННОТАЦИЯ

В исследовании на крысах установлено, что применение нового производного 2-оксоиндолина с диуретическими свойствами – индолинорена – при экспериментальном гемодинамическом отеке легких уменьшает степень гидратации легочной ткани. По сравнению с гипотиазидом, индолинорен оказывает более выраженный противоотечный эффект, о чем свидетельствует достоверное уменьшение легочного коэффициента и увеличение сухого остатка легочной ткани при экспериментальном отеке легких.

Ключевые слова: диуретики, отек легких, легочной коэффициент.

УДК 616.24-005.98:547.757

Т.І. Тюпка, іна, , А. О. Мінаєва

Національний фармацевтичний університет, м. Харків

ВПЛИВ ІНДОЛІНОРЕНУ НА СТУПІНЬ ГІДРАТАЦІЇ ЛЕГЕНЕВОЇ ТКАНИНИ ПРИ ЕКСПЕРИМЕНТАЛЬНОМУ НАБРЯКУ ЛЕГЕНЬ

АНОТАЦІЯ

У дослідженні на щурах встановлено, що застосування нового похідного 2-оксоіндоліну з діуретичними властивостями – індолінорену – при експериментальному гемодинамічному набряку легень зменшує ступінь гідратації легеневої тканини. У порівнянні з гіпотіазидом, індолінорен чинить більш виражений протинабряковий ефект, про що свідчить достовірне зниження легеневого коефіцієнту та збільшення сухого залишка легеневої тканини при експериментальному набряку легень.

Ключові слова: діуретики, набряк легень, легеневий коефіцієнт.

Відомості про авторів:

Тюпка Тетяна Іванівна — д. мед. н., професор каф. патологічної фізіології НФаУ

e-mail: *****@***ru

Кононенко Надія Миколаївна — д. мед. н, професор, зав. каф. патологічної фізіології НФаУ;

Маркіна Анна Юріївна — здобувач каф. патологічної фізіології НФаУ;

Мінаєва Аліна Олексіївна — аспірант каф. патологічної фізіології НФаУ.