CC BY
9"BicHUK стоматологИ", № 1, 2016
УДК 57.083.3 - 035.2:(612.176+599.323.4)
S. A. Shnaider, M.D.
SE "the Institute of Stomatology of the National Academy of Medical Sciences of Ukraine"
THE STUDY OF THE ADAPTOGEN PECULIARITIES OF THE PREPARATION FROM VEGETATIVE POLYPHENOLS AT THE RESTORATION OF ACUTE EMOTIONAL PAINFUL STRESS
ABSTRACT
The aim of study is the studying protective properties preparation of plant polyphenols phitosed and tranquilizer diazepam in conditions of manifestations emotionally-algesic stress of rats. Materials and methods. The experiment was carried out on 5-month-old 24 male rats. They were devided into 4 groups (6 rats per group). The first group was intact; second, third and fourth groups were in conditions of acute emotionally-algesic stress. The rats were given per os : second group - water, third group - diazepam at a dose 1,25 mg/kg body weight of rats; fourth group - phitosed at a dose 1,0 ml/kg (at a dilution of 1:1) for 60 minutes before of stressing.
Conclusions. Researches showed stress-protective effects of di-azepam and strongly pronounced protective adaptogenic properties of phitosed in conditions of acute emotionally-algesic stress of rats.
Key words: acute emotionally-algesic stress, plant polyphenols, phitosed, diazepam, adaptogenic effects.
С. А. Шнайдер
Государственное учреждение «Институт стоматологии Национальной академии медицинских наук Украины»
ИЗУЧЕНИЕ АДАПТОГЕННЫХ СВОЙСТВ ПРЕПАРАТА РАСТИТЕЛЬНЫХ ПОЛИФЕНОЛОВ В УСЛОВИЯХ ВОСПРОИЗВЕДЕНИЯ ОСТРОГО ЭМОЦИОНАЛЬНО-БОЛЕВОГО СТРЕССА
В экспериментах на 24 белых крысах изучено влияние препарата растительных полифенолов (ПФ) - фитоседа и стандартного транквилизатора диазепама на показатели периферической крови, состояние тест-органов и ульцеро-генные эффекты при воспроизведении острого эмоционально-болевого стресса (ЭБС) у крыс. Установлены стресс-протективые эффекты диазепама и ярко выраженные защитные адаптогенные свойства препарата фи-тосед в условиях воспроизведения острого эмоционально-болевого стресса.
Ключевые слова: острый эмоционально-болевой стресс, растительные полифенолы, фитосед, диазепам, адапто-генные эффекты.
С. А. Шнайдер
Державна установа «1нститут стоматологи Нацюнально! академй медичних наук Украши»
ВИВЧЕННЯ АДАПТОГЕНИХ ВЛАСТИВОСТЕЙ ПРЕПАРАТУ РОСЛИНИХ ПОЛ1ФЕНОЛ1В В УМОВАХ В1ДТВОРЕННЯ ГОСТРОГО ЕМОЦШНО-БОЛЬОВОГО СТРЕСУ
крот, стан тест- оргатв та ульцерогент ефекти в умовах вiдтворення гострого емлоцшно-больового стресу (ЕБС) у щурiв. Встановлею стрес-протективш ефекти дiазепаму та яскраво виражем захисн адаптоген властивостi препарату фтосед в умовах вiдтворення гострогь емоцтно-больового стресу.
Ключовi слова: гострий емоцшно-больовий стрес, рослинш полiфеноли, фтосед, дiазепам, адаптоген ефекти.
The stomatological intrusions, accompanied by the emotional painful stress (EPS), result in inhibition of the regenerative processes in oral tissues, and the use of the currently in use sedative preparations-anxiolytics does not always eliminate its consequences [1].
Lately it was determined, that polyphenols (PP) of vegetative origin take part in general and local resistibility of organism, being the substitutes of hormones, humoral controllers, etc. [2].
The representatives of the main classes and groups of polyphenol compounds display anti-inflammatory effect. The flavonoids quercethin, rutin, kaempferol and their glycosides are the famous anti-inflammatory preparations. The anti-inflammatory activity is observed in baicalein, apigenin, luteolin and their glycosides [3]. In nature fla-vonoids are mostly found in higher plants and are met in the form of glycosides, rarer - aglycons.
In this connection the PP from plants attract attention of the researchers as the promising substances for the production of medical preparations.
The aim of this investigation. The study of the protective characteristics of the preparation from vegetative polyphenols - phytosed and tranquilizer diazepam at display of acute emotional and painful stress in rats.
The materials and the methods. The experiment was held with 24 white he-rats of 5 months old of Vistar line of herd breeding. The influence of Phytosed SC "Effect", Kharkov, Ukraine) and standard anxiolytic Diazepam ("Zdorovye Narody", LTD, Kharkiv, Ukraine) upon the indices of periphery blood, the state of test-organs and ulcerogenic effects at the restoration of acute emotional painful stress (EPS) by Desiderato method [4].
All animals (6 in each group) were kept to standard diet of vivarium: the 1st group - intact one; in rats from the 2nd- 4th groups the acute EPS was restored. 60 minutes before the stressing the rats were given per os the following: the animals from the 2nd group - water; the 3rd one -diazepam (seduxen) dosed at 1,25 mg/kg of rat's weight; the 4t group - phytosed dosed at 1.0 ml/kg (in solution 1:1).
Phytosed is the complex of vegetative polyphenols, possessing sedative, anxiolytic characteristics. It contains the extracts from the fruits of hawthorn, coriander, oats; herbs of Melissa, melilot, motherwort; multiple fruit of hop. The biologically active substances of phytosed are polyphenols - flavonoids, coumarins, phenol acids and essential oils. The flavonoids - quercethin, quercetrin, vitexin, glycosides of apigenin, luteolin and others - are the most studied among them.
В docnidax на 24 быих щурах вивчено вплив препарату рос-линних полiфенолiв (ПФ) - фтоседу та стандартного транквiлiзатору - дiазепаму на показники перiферично'í
© Shnaider S.A., 2016.
"BicHUK cmoMamojioziï", № 1, 2016
Table 1
The influence of the investigated preparations on the cellular contents of periphery blood in rats at the restoration of acute EPS in rats (M±m; p; р1)
Groups of animals The investigated preparation blood corpuscles
Neutrophils Eosinophils (%) Monocytes (%) Lymphocytes (%) Leucocytes (g/l)
banded segmented
Intact 2,2±0.9 25,4±1,6 4,4±1,1 6,2±1,2 71,8±4,2 13,9±0,7
EPS +water (control) 3,8±0,3 67,5±16,9 p=0,03 2,3±0,2 p=0,09 4,5±1,8 22,5±4,3 p<0,001 14,7±0,4
+ diazepam 2.7±0,4 p1=0,05 62,8±2,3 p<0,001 2,5±0,7 6,5±0,9 26,3±2,4 p<0,001 10,8±0,5 p=0,004 p1<0,001
+ phytosed 2,7±0,8 56,3±4,2 p<0,001 4,0±1,1 5,5±1,4 32,3±5,2 p=0,002 10,3±2,3
No te . In tabl. 1-3 the certainty index p is calculated related to the intact group; p1 - control one.
Table 2
The influence of the investigated preparations upon the relative mass of organs of rats at the restoration of acute EPS in rats (M±m; p; р1)
Investigated preparations Groups of animals
Intact EPS
+water (control) +diazepam +phytosed
Relative mass of organs Adrenal glands 26,7±1,8 22,5±1,3 p=0,08 24,9±1,1 26,7±1,5 p1=0,06
Spleen 421±37,7 286±32.2 p=0,02 342±30,9 312±22,4 p=0,03
The acute EPS of anxious expectation of painful affection was made by 3-hour keeping of rats in cages of Desiderato construction, in which direct current feed of 56 mA was performed with random intervals of time either for half of the cage, or for the platform over the floor [4].
After the completion of the stressing in rats the periphery blood was taken intravitally from the caudal vein and the number of neutrophils, eosinophils, monocytes, lymphocytes, the total number of leucocytes, were estimated.
The animals were taken out of the experiment by total phlebotomy from heart, executed under thiopental narcosis (40 mg/kg). After the slaughter in animals the relative mass of adrenal glands and spleen [organ mass (mg)/rats mass (g)] x100, as well as ulcerogenic effect, were determined. Stomach was extracted, opened by lesser curvature, washed, dried with tissue and the thickness of the wall of mucous coat of stomach (MCS) was measured. Then, the number of injuries, appeared at MCS, was calculated, their linear size (in mm) was measured with trammel. The frequency of injuries of MCS was revealed (in %), also the multiplicity - the quantity of erosions and ulcers. The graveness of injuries of MCS was estimated according to 4-points system: up to 1mm - 1 point, 1-2.9 mm - 2 points, 3-4,9 mm - 3 points, 5 mm and more - 4 points.
The results, achieved at investigation, were processed by standard method with determination of t-criteria of the significance of differences by Student.
The findings. The life-time study of the periphery
blood corpuscle in rats after the restoration of EPS in them has shown the considerable shifts in its cellular contents. So, the number of segmented neutrophils grew by 2.7 times (p=0,03) and the number of lymphocytes shortened by 3,2 times (p<0,001; tabl. 1).
Under the influence of diazepam, introduced simultaneously to acute EPS, the number of stab neutrophils, eosinophils and monocytes normalized. The number of segmented neutrophils and lymphocytes remained almost at the level of these control groups. Diazepam simultaneous to stress caused the true shortening of the general number of leucocytes (G/l) in comparison to the control (pj<0,001) and intact (p=0,004) groups (tabl. 1).
The results of the studies of the cellular contents of blood of rats under the influence of vegetative PP of the preparation phytosed have revealed the following. The number of stab neutrophils, eosinophils, monocytes and the general number of leukocytes under the influence of phytosed do not differ considerably from the data from intact groups. The number of segmented neutrophils and lymphocytes changed uncertainly in comparison to the data of control groups. But then the number of lymphocytes reduced by 2,2 times (p=0,002), and segmented neu-trophils grew (p<0,001) relative to the intact group (tabl. 1).
The findings, given in the table 2, speak of the fact that the acute EPS caused the considerable decrease of the relative mass of spleen and the tendency of the reduction of mass of adrenal glands. These phenomena correspond to the commonly known notions on the after-effects of the
"BiCHUK cmoMamonoziï", № 1, 2016
restored stress.
In the group with the introduction of diazepam simultaneous to stress the relative mass of test-organs of rats did not differ essentially from such of intact animals. Under the influence of phytosed the relative mass of adrenal glands grew in comparison to the control group (pi=0,06) and achieved the level of intact one (table 2). So, phytosed has normalized the relative mass of adrenal glands of the stressed rats. At that the relative mass of spleen was really lower, than in intact group.
The results of the morphometric studies of the mucous coat of stomach for the determination of the ulcerogenic effect are presented in the table 3.
The studies have shown, that acute EPS caused
the real narrowing of the mucous coat of stomach of rats (by 1,3 times; p=0,03) (tabl. 3). The number of erosions under the influence of EPS increased by 2,3 times (tendency; p=0,08), and the number of ulcers -by 20,3 times (p=0,02). The acute EPS caused 100 % frequency of MCS of rats in comparison to 5,7 % in intact group. After the restored stress the appearance of the great number of large erosions, also ulcers on mucous coat of stomach, were observed. Thus, the graveness of affections with erosions (in points) grew trice, and with ulcers - by 24,3 times (tabl. 3).
Table 3
The influence of the investigated preparations on the displays of ulcerogenic effect at restoration
of acute EPS in rats (M±m; p; р1)
Groups of animals
Investigated indices Intact Emotional painful stress
+water (control) +diazepam +phytosed
Thickness of MCS 1,20±0,07 0,96±0,08 p=0,03 0,96±0,09 p=0,05 0,91±0,07 p=0,011
Frequency of affections (%) 5,7 100 85,7 50
Erosions 1,1±0,2 2,5±0,7 2,1±1,0 1,5±0,7
Number of p=0,08
Ulcers 0,4±0,3 8,1±2,9 p=0,02 5,4±2,2 p=0,04 3,8±2,3
Graveness of af- Erosions 2,1±0,5 6,3±2,1 5,3±2,4 3,8±1,8
fection p=0,08
(in points) Ulcers 0,6±0,3 14,6±3,3 p=0,001 9,9±4,6 p=0,07 5,1±3,3 P1=0,07
Diazepam, introduced simultaneously to the acute EPS, reduced by 14,3 % the frequency of injuries on mucous coat of stomach in comparison to 100 % in group of the stressed rats. The number and graveness of the affection with ulcers reduced by 1,5 times in comparison to the control group (p>0,05). At that the number of ulcers and the graveness of affection with them under the influence of diazepam were considerably higher, than in intact groups. This fact speaks of insufficient prevention by this preparation of the consequences of the restored acute EPS.
The preparation of the vegetative PP - phytosed reduced the frequency of the affection of MCS of rats compared to the control group by 50 %. The quantity of ulcers on MCS under the influence of phytosed twice as shortened (p>0,05), and the graveness of ulcer injuries - reduced by 2,9 times (tendency; pi=0,07) in comparison to the data of control groups. The number of erosions corresponded to this index in intact animals.
So, the held investigations have shown stress-protective effects of diazepam and vivid protective adaptogenic characteristics of the preparation from vegetative polyphenols - phytosed at the restoration of acute emotional painful stress in rats.
The conclusions. 1. Diazepam normalized the relative mass of test organs (adrenal glands and spleen) at display of acute emotional painful stress.
2. Phytosed had positive influence on the indices of periphery blood of stressed rats.
3. Phytosed has revealed the protective characteristics versus the mucous coat of stomach of stressed rats, at that the degree of their displays was much higher, than the ones of standard tranquilizer - diazepam.
REFERENCES
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2. Levitskiy A.P., Voskresenskiy O. N., Nosiychuk S. V. The role of food polyphenols in the formation of the local non-specific resistance of oral tissues. Visnikstomatologii. 2005;3:2-8.
3. Theoharides T. S., Alexandrakis M. Anti-inflammatory actions of flavonoids structural reguirements for new design. Int. J. Immupathol. Pharmacol. 2001;14:119.
4. Desiderato O., Kinnon J., Hisson H. Development of gastric ulcers in rats following stress termination. J. Comp. Physiol. Psyhol. 1974;87:208-214.
Acted 05.01.16
