THE FEATURES OF ARTERIAL THYMUSATNALBUPHINEACTION

The article presents and analyzes the results of the pilot study, at which opioid nalbuphine was daily administered in white rats-males of reproductive age (1,5–3,5 – month) for six weeks and one week after discontinuation of therapy. The drug “nalbuphine” was injected intraperitoneally daily 1 per day over 42 days. The data were compared with a control group of intact animals. The study was conducted on 52 animals, which were divided into eight groups. Morphometric study was performed using a system of visual analysis of histological preparations. Dynamics of changes in the external and internal diameters of medium caliber arteries thymus, media and lumen area, Vohenvorta ratio was studied. The increase of the external diameter and reduction of the internal diameter of the medium caliber arteries, increase of the area of media, increase of the area of the lumen, increase of the Vohenvortaratio.The blood vessels are dilated, gaps filled elements of the blood vessels around the swelling, the process of vascular sclerosis.


Introduction
Urgent social and medical issues of our time are opioid addiction [1][2][3]. Opioids have been used in medicine for therapeutic purposesfor a long time. Representative opioid is nalbuphine (nubain) a narcotic analgesic, semi-synthetic opioid derived phenanthrene [4].
The effect of opioids on the organs of the immune system is insufficiently studied, particularly at primary lymphoid organs, the thymus. In the special literature there are data about influence on the structural organization of the thymus various harmful factors [5,6] such as indomethacin, histynat, methyl alcohol, naloxone, hyperosmolar solution [7], imunofan [8], cyclophosphamide [9][10][11] but there are only a few data on the impact of drugs in this important primary lymphoid organs. Application of imunofan has the pronounced effect on the maturation and differentiation of thymocytes in the thymus of rats of reproductive age [12]. After 1 day after administration of immunosuppressive cyclophosphamide there takes place the breachof hematotymusnoho permeability barrier [13,14]. Within 30 days after administration of cyclophosphamide differences in the structure of rat thymus substantially leveled [15]. The data on the effect of opioids on the cerebellum, pancreas, tongue, eyeball, skin, cerebrumwere published [16][17][18].
The thymus as the primary organ of the immune system provides antigen independent proliferation and differentiation of a subpopulation of T lymphocytes. The exterior is covered with a connective tissue capsule that gives partitions into the parenchyma, dividing it into segments. Each particle consists of cortex and medulla.

Aim of the research
To study the dynamic changes of structural parameters of medium caliber arteries thymus during a six-week action of nalbuphine experiment.
Experimental animals were divided into 8 groups of 5 animals ( Table 1). 12 white males served as the control rats thatwere injected with 0.9 % sodium chloride solutioninstead of nalbuphine.
Morphometric study was performed using a system of visual analysis of histological preparations. Image of histological preparations on a computer monitor was taken out of the microscope MICROmed SEO SSAN using the camera Vision CCD Camera. Morphometric study was conducted using software VideoTesT-5.0 KAAr Image Base and Microsoft Excel on a personal computer in the histological preparations stained with hematoxylin and eosin. We determined the outer and inner diameter of medium caliber arteries thymus, the average area of membrane and lumen of the vessel, the Vohenvortaratio. Digitally statistically processed values presented the average and standard deviation (M±m). Probability differences of average values (p) were determined by Student's test. Differences were considered likely at p<0,05.
The drug "nalbuphine" was daily injected intraperitoneally (10-11 am) 1 per day over 42 days (6 weeks). Every week dose of nalbuphine injections was increased ( Table 1) according to recommendations [19]. Based on the cooperation agreement between the Department of Human Anatomy and Histology of the Medical Faculty SHEE "Uzhhorod National University" and the Department of Anatomy Lviv National Medical University named Danylo Galician, research conducted at the Lviv National Medical University. D. Galician All the animals were kept in vivarium conditions of Lviv National Medical. Experiments were conducted under the provisions of the "European Convention for the protection of vertebrate animals used for experimental and other scientific purposes" (Strasbourg, 1986) [20], Council Directive 86/609 / EEC (1986) [21], the Law of Ukraine №3447-IV "On protection of animal" treatment common ethical animal experimentation adopted the first national Congress on bioethics Ukraine (2001) [22].

Results
The dynamics of changes in the external and internal diameters of medium caliber arteries was studied. The gradual increaseof external diameter in maximum 5 weeks and of internal diameter in 1 week was revealed. Further performance decreases ( Table 2).
Results of Table 1 are shown schematically in Fig. 1. We can see the dynamics of internal and external diameters of medium caliber arteries during the experiment.  The dynamics of changes in the area of media and lumen area, Vohenvorta ratio in arteries of medium caliber. There was revealed a gradual increase, with a maximum of 5 weeks in the area of media and after 2 week in the lumen area. Further performance decreases (Table 3). 50 Table 2 is shown schematically in Fig. 2. You can see the dynamics of the media area and lumen area of medium-caliber arteries during the experiment.  On histological preparations were analyzed the microscopic changes in thymic medulla slices of intact white-male rats (Fig. 4, a) before and after 1 week of introduction of nalbuphine (Fig. 4, b). No significant changes were detected. There was analyzed the microscopic state of the cortex slices thymus ofanimals after three weeks of the experiment. The discovered was expanded and filled with thebloodveins (Fig. 5).
After five weeks of the experiment were revealed many microscopic changes in the cortex slices thymus of animals. Plot thickened interlobular connective tissue, indicating leukocyte diapedesis through the wall of venules; expanded and filled with blood vessels (Fig. 6).
A week after discontinuation microscopic changes in the thymic cortex slices of animals are not returned to normal. Pathological process, developed in the thymus was not come back (Fig. 7). 290  (2) and filled with blood vein (3); perivascular edema (4); 5 -Tlymphocytes. Stained with hematoxylin and eosin Coll.: lens×20, glasses×15

Discussion
At the one-week opioid nalbuphine exposure to male rats of reproductive age the was significantly increased the outer diameter of arteries of medium caliber 5,1 % and 3,3 % inner diameter. After 2 weeks, the external diameter decreases by 0,6 % and the internal diameter by 0,8 %. After 3 weeks of daily administration of nalbuphine outer diameter decreases by 0,4 % and an internal diameter by 12,5 %, due to the swelling of endothelial cells. Within 4 weeks of the experiment is the growth of the external diameter of the arteries by 3,6 %, but inside diameter further reduced (to 0,9 %). After 5 weeks of the experiment the outer diameter increases (2,5 %) and domestic one (3,4 %). This can be explained by the loosely placed fibers of connective tissue, edema and sclerosis. After 6 weeks of daily administration of opioid indicator external diameter decreases by 0,6 % and domestic by 6,9 %. After 1 week after discontinuation of the drug outside diameter arteries decreased by 5,3 % and is higher than this indicator in intact animals of 4,3 %, and the inner diameter is reduced slightly (by 0,2 %) and is lower than in intact animals to 14,8 % (Table 2, Fig. 1).
After a week of opioid nalbuphine action in the body of male rats of reproductive age significantly (p<0,05) increases the average area of membrane (media) medium caliber arteries by 9,2 % and lumen area by 6,2 %. After 2 weeks, media area increased by 1,4 % and lumen area by 0,9 %. After 3 weeks of daily administration of nalbuphine media area increased by 2,6 % and the lumen area decreased by 2 %. Within 4 weeks of the experiment area media increased by 6,1 %, due to the thickness of basement membrane, lumen area decreased by 1,5 %. After 5 weeks of the experiment the area of media increases (1,7 %) and lumen area (by 2,1 %). After 6 weeks of daily administration of opioid media area index decreased by 1,2 % and lumen area by 0,9 %. After 1 week after discontinuation media area decreased by 3,2 % and is higher than this indicator in intact animals of 17,4 % and lumen area decreased slightly (by 0,6 %) and is lower than in intact animals 4 % (Table 3, Fig. 2).
These structural changes in the walls of arteries medium caliber prove the dynamics of Vohenvorta ratiochange (Table 3, Fig. 3).

Conclusions
As a result, the impact of opioid nalbuphine on male rats of reproductive age for six weeks revealed: 1) increased external diameter arteries of medium caliber 10.1 %; 2) reduced internal diameter arteries of medium caliber 14.6 %; 3) increased area of media medium caliber arteries by 21.3 %; 4) increased area of the lumen of medium caliber arteries by 4.7 %; 5) Vohenvorta ratio increased by 11.3 %; 6) gradual increase in the area of media medium caliber arteries thymus with maximum 5 weeks by 22 %. Then the media area begins to decrease; 7) the blood vessels are dilated, gaps filled with elements of the blood vessels around the swelling; 8) the process of sclerosis of blood vessels and circulatory hypoxia. After one week after discontinuation of the drug: 1) the outer diameter of the arteries decreases, but remains higher than the indicator in intact animals of 4.3 %; 2) internal diameter decreases, but remains lower than in intact animals to 14.75 %; lumen area decreases, but remains lower than in intact animals by 4 %; 3) the area of media medium caliber arteries thymus begins to decrease but is higher by 17,4 % than in intact animals.