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Ткачова О.В. Вплив гелю «Живітан»® на біохімічні показники крові в умовах гострого тромбофлебіту вени вуха у кролів
14.05.2014, 09:26

Резюме
Ткачова О.В. Вплив гелю «Живітан»® на біохімічні показники крові в умовах гострого тромбофлебіту вени вуха у кролів.
За результатами біохімічних досліджень в умовах експериментального гострого тромбофлебіту встановлений позитивний вплив гелю «Живітан»® на стабілізацію мембран, зменшення запального процесу, відновлення показників ліпідного та білкового обміну. Порівняно з референс-препаратами дія гелю «Живітан»® проявлялася швидше і була більш виразнішою.
Ключові слова: фармакологічне дослідження, тромбофлебіт, гель «Живітан»®.
Резюме
Ткачева О.В. Влияние геля «Живитан»® на биохимические показатели крови в условиях острого тромбофлебита вены уха у кроликов.
По результатам биохимических исследований в условиях экспериментального острого тромбофлебита установлено положительное влияние геля «Живитан»® на стабилизацию мембран, уменьшение воспалительного процесса, восстановление показателей липидного и белкового обмена. По сравнению с референс-препаратами действие геля «Живитан»® проявлялось быстрее и было более выраженным.
Ключевые слова: фармакологическое исследование, тромбофлебит, гель «Живитан»®.
Summary
Tkachevа O.V. Impact of «Zhyvitan»® gel on biochemical parameters of blood in the presence of acute thrombophlebitis of ear vein in rabbits.
According to the results of biochemical studies in experimental acute thrombophlebitis found a positive effect of the gel «Zhyvitan»® at stabilizing the membranes, reducing inflammation, restoring lipid and protein metabolism. Compared to the reference-preparations action gel «Zhyvitan»® was quicker and more pronounced.
Key words: pharmacology study, thrombophlebitis, gel «Zhyvitan»®.

УДК 615.272.4:616.14-002:615.454

 

Національний фармацевтичний університет (Харків)

 

Национальный фармацевтический университет (Харьков) 61002, г. Харьков, ул. Пушкинская, 53
National University of Pharmacy (Kharkov) Ukraine, 61002, Kharkov, 53, str. Pushkinska

 

tkachevaov@gmail.com

Introduction. Chronic venous insufficiency (CVI) of the lower extremities is the most common peripheral vascular disease. Number of patients with this pathology increases worldwide, despite the variety of treatments. Various manifestations of venous insufficiency are found in 50-70% of the population, but only 8% of patients receive timely treatment [1]. This situation leads to the fact that CVI assumes the character of an important medical problem because the consequences of delayed medical care are complications of venous diseases - trophic ulcers and pulmonary embolism, requiring long and quite expensive treatment [2].

Treatment of CVI should be combined, aimed at all pathogenetic links through a combination of conservative methods (compression therapy, physical therapy, drug therapy) and surgical treatment [2].

Conservative treatment of CVI is crucial, since surgical intervention, which is applied to 10% of patients, provides no guarantee of recurrent relapse. In the combined treatment of CVI venotropic drugs, that reduce venous pressure in varicose veins, strengthen capillaries and improve the blood supply of the affected tissues, are very important [4].

Modern combined vein protective remedies should not only normalize the structure and function of the venous wall, but also stimulate lymphatic drainage, improve rheological properties of blood and microcirculation, suppress inflammatory processes in the soft tissues of the extremities, improve trophism and stimulate tissue repair. The combination of different types of pharmacological action is only possible in case of the creation of combined drugs.

Taking into account current trends of CVI pharmacotherapy, staff of the Central Research Laboratory of the National University of Pharmacy together with scientists from JSC “CPP” Red Star” created a new combination drug – “Zhyvitan”® gel. The “Zhyvitan”® gel contains an extract of horse chestnut seed and an extract of comfrey root. The combination of two extracts will enhance pharmacodynamics of the drug and indications for use: both for a topical treatment of CVI, and for the prevention and treatment of trophic ulcers.

The aim of this work is to study the impact of “Zhyvitan”® gel on biochemical parameters of blood on the model of acute thrombophlebitis of ear vein in rabbits.

Materials and methods of study. The experiment was carried out on underbred male rabbits weighing 2,0-2,5 kg. To reproduce thrombophlebitis a ligature was applied to a marginal vein of the rabbits’ ears, after previous depilation and disinfection, and above it the vein cavity was injected with 0,2 ml of Lugol's solution. The ligature was removed in 2 hours [3]. The model allowed to obtain blood flow disturbance due to stasis and inflammation of the venous wall, which is typical for most thrombotic states in humans.

In the experiment 30 rabbits divided into 6 groups of 5 animals in the group were used: 1 – intact control; 2 – positive control – untreated animals with reproduced pathology; 3 – animals that in the setting of thrombophlebitis were treated by “Zhyvitan”® gel; groups 4 – 6 – animals that in the setting of thrombophlebitis were treated by reference drugs. As comparator drugs, soft drugs of “Dr. Theiss Naturwaren GmbH” company (Germany) were used: 1) “Venen Theiss Gel” containing 3% extracts of horse chestnut seed and 0.4% calendula blossoms; 2) “Dr. Theiss Comfrey Gel” containing 5% comfrey root extract; 3) “Dr. Theiss Comfrey Ointment” containing 10% comfrey root tincture and 1% Vitamin E.

The study medication and comparator drugs were applied a thin coat to the affected area of the rabbits’ ears twice a day (in the morning and in the evening) in a conventional therapeutic dose of 930 mg/kg, which was established in previous studies [5]. Removal of animals from the experiment was conducted on the 10th day after the development of thrombophlebitis by air embolism. Parameters to assess the overall status of animals and the impact of drugs on the course of pathology were selected based on the pathogenesis and clinical manifestations of thrombophlebitis.

Assessment of clinical parameters and the pharmacological action of drugs was carried out on the basis of measurements of the dynamics of thrombus length. In order to confirm systemic development of pathological process on the 7th and 10th day in blood serum using a set of “BIOLA-TEST” reagents the biochemical parameters: aspartate aminotransferase (AST), total protein, urea, total lipids, were studied.

All the interventions and euthanasia of animals were carried out in compliance with the ethical code (Strasbourg, 1986). The results obtained were processed by Statistica 6.0 program based on analysis of variance for data with repeated measurements using Kruskal-Wallis criteria and Newman-Keuls criteria at the level of probability р< 0,05.

The results and their discussion. The pathology reproduction in animals was accompanied by the development of thrombosis of the ear marginal vein. The thrombus length in the beginning of observations was about 49 mm in animals of all experimental groups, indicating the hemodynamic abnormalities in animals (Fig. 1).

Under the influence of “Zhyvitan”® gel the thrombolysis rate increased – the size of thrombosed areas beginning from the 6th day of treatment was significantly lower than the findings in PC group. When compared with the reference drugs the thrombolytic action of “Zhyvitan”® gel was at the level of the drug “Dr. Theiss Comfrey Ointment” and according to the speed of action it had a slight advantage over the drugs “Venen Theiss Gel” and “Dr. Theiss Comfrey Gel”.

 

Table 1

Impact Gel "Zhyvitan» ® products and comparing the length of the thrombus
in acute vein thrombosis in rabbits ears, M±m

Length of thrombus, mm

Positive control

“Zhyvi-tan”® gel

“Venen Theiss Gel”

“Dr. Theiss Comfrey Gel”

“Dr. Theiss Comfrey Ointment”

24 hours

48,6±1,6

48,8±1,9

48,2±1,5

48,6±1,4

49,2±1,7

4 day

40,8±2,1

34,8±3,6

39,2±1,7

37,8±0,6

28,6±3,3*

6 day

35,8±2,3

27,2±2,9*

34,6±0,8

31,2±0,7

22,6±3,2*

8 day

28,4±2,5

18,8±2,6*

20,2±1,4*

21,6±2,2*

18,0±2,1*

10 day

22,2±2,6

13,8±1,5*

15,6±1,4*

14,0±1,8*

13,6±0,5*

Note: * – differences are likely related to the group of positive control, р<0,05.

 

The development of thrombophlebitis led to systemic inflammation and expressive intoxication, which was reacted by the liver. In animals, the level of intracellular enzyme AST increased due to membrane-destructive processes in the liver. The development of thrombophlebitis also led to certain changes in the metabolism of animals: probably, the level of total protein reduced, the level of urea and total lipids in blood serum increased (Table 2).

In the group of animals treated with “Zhyvitan” ® gel AST level remained at the level of intact control during the entire period of study and was significantly higher than PC group findings, indicating an expressive membrane stabilizing and vein protective effect of the drug.

Applying of “Zhyvitan” ® gel on the 7th day contributed to a significant increase of total protein and reduction of urea level and total lipids to the level of intact control.

Thus, the results of biochemical studies showed the positive impact of “Zhyvitan” ® gel on membrane stabilization, reduction of the inflammatory process, restoration of lipid and protein metabolism.

 

Table 2

Effect of drugs on biochemical indices in acute vein thrombosis in rabbits ears, M±m

Рarame-ters

Days
expe-riment

Groups of animals

Intact control

Positive control

“Zhyvi-tan”® gel

“Venen Theiss Gel”

“Dr. Theiss Comfrey Gel”

“Dr. Theiss Comfrey Ointment”

AST, mmol / h*l

0

0,25±

0,02

0,23±

0,01

0,25±

0,02

0,26±

0,02

0,25±

0,02

0,25±

0,02

3

0,24±

0,03

0,27±

0,01

0,19±

0,02**,

***

0,31±

0,03*

0,26±

0,06

0,29±

0,01*

7

0,24±

0,03

0,31±

0,02*

 

0,19±

0,03**, ***

0,31±

0,02*

 

0,25±

0,03

 

0,29±

0,02*

 

Total protein, g/l

0

75,89±

3,61

75,97±

0,13

77,92±

2,31

74,97±

1,16

73,03±

2,54

76,35±

2,93

3

75,79±

2,24

69,94±

0,35*

72,15±

0,54**

70,94±2,52

69,78±

1,23*

70,85±

1,93*

7

76,30±

3,67

71,10±

1,56*

81,13±

1,29**, ****

75,34±

2,17

71,28±

1,16*

73,72±

2,80

Urea,

mol/l

0

3,90±

0,41

3,85±

0,26

3,68±

0,48

4,05±

0,39

3,36±

0,18

3,56±

0,55

3

3,69±

0,34

6,92±

0,66*

6,34±

0,74*

6,65±

0,67*

6,63±

0,49*

5,78±

0,70*

7

3,67±

0,37

6,23±

0,49*

4,73±

0,35**

5,11±

0,49*

4,96±

0,20**

5,23±

0,42*

Total lipids,

g/l

0

3,10±

0,27

3,25±

0,47

3,56±

0,49

3,42±

0,44

3,50±

0,28

3,35±

0,22

3

3,01±

0,29

6,0±

0,62*

6,33±

0,34*

5,78±

0,58*

6,22±

0,14*

5,28±

0,49*

7

3,67±

0,37

6,10±

0,46*

4,40±

0,27**

4,73±

0,84

4,14±

0,28**

4,58±

0,50

Notes:

1. * – differences are likely related to the group intact control, р<0,05;

2.   ** – differences are likely related to the group of positive control, р<0,05; 

3. *** – probable differences regarding drug Venen Theiss gel, р<0,05;

4. **** – probable differences on gel comfrey Dr. Theiss, р<0,05.

 

In the presence of acute thrombophlebitis an expressive anti-inflammatory, membrane stabilizing and antithrombotic activity of “Zhyvitan” ® gel according to the effect on the dynamics of hematological parameters: fibrinogen, leukocytes, prothrombin time and clotting time, was found [5].

This dynamics of biochemical and hematological parameters indicates almost complete lysis of thrombus and restoration of the metabolism in the system under the influence of “Zhyvitan” ® gel. Compared with the reference drugs the effect of “Zhyvitan” ® gel manifested faster and was more expressive.

Conclusions

1. “Zhyvitan” ® gel in the presence of acute thrombophlebitis contributed to the expressive acceleration of thrombolysis and normalization of blood biochemical parameters.

2. Compared with the reference drugs the effect of “Zhyvitan”® gel was expressive and faster, which will allow to use efficiently the drug for topical treatment of acute and chronic venous insufficiency.

Література
1. Богачев В.Ю. Начальные формы ХВН нижних конечностей: эпидемиология, патогенез, диагностика, лечение и профилактика [Електронний ресурс] / В.Ю. Богачев // Consilium–Medicum. – 2004. – Т. 6, № 4. – Режим доступу до журналу: http://www.consilium-medicum.com/article/10188.
2. Мишалов В.Г. Лечение ХВН нижних конечностей / В.Г. Мишалов, Н.Ю. Литвинова // Серце і судини. – 2010. – № 3. – С. 29–35.
3. Спосіб моделювання тромбофлебітів периферичних судин: Інформаційний лист НФаУ від 20.11.2002 р. / Л.М. Малоштан, О.В. Должикова, І.О. Батура. – К.: Укрмедпатентпром, 2002. – 2 с.
4. Стойко Ю.М. Патогенетические аспекты консервативной терапии ХВН у беременных [Електронний ресурс] / Ю.М. Стойко, А.В. Замятина // Consilium–Medicum. – 2007. – Т.9, №6. - Режим доступу до журналу: http://www.consilium-medicum.com/medicum/article/13397.
5. Ткачова О.В. Дослідження венопротекторної дії гелю «Живітан®» на моделі гострого тромбофлебіту вени вуха у кролів / О.В. Ткачова, Л.В. Яковлєва // Український медичний альманах. – 2012. – Т. 15, № 5. – С. 263–265.

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