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A Study on Rat Cranial Blood Vessels Under the Influence of Ethanol

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Published: 8th Apr 2021

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Morphological changes of cranial venae cavae of rats in the early postnatal ontogenesis under the influence of ethanol

Abstract

Background: Morphology of major arterial and venous lines at different ages, and especially their changes under the experimental exposure gives a deeper understanding of problems in the structure of blood vessels in normal and pathological conditions. Aim: To study the changes of intrapericardial cranial venae cavae of rats in postnatal development under the influence of ethanol.

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Methods and materials: Investigated intrapericardial part of the right (RCVC) and left (LCVC) cranial venae cavae in 120 rats 6, 11, 16, and 22 to 30 days of ages. 60 of them received ethanol with mother’s milk. For this females were daily intragastrically administered 40 ° ethanol in a dose of 8 ml / kg during the lactation period (22 days). The control group consisted of 60 rats of the same age, females – mothers who daily intragastrically received distilled water in the same dose. Results: It is established that under the influence of ethanol decreases the thickness of the wall of the cranial vena cava. Thickening of the collagen and elastic fibers, reticular fibers and destruction of myocytes observed. Conclusion: To study the changes of intrapericardial cranial venae cavae of rats in postnatal development under the influence of ethanol.

Keywords: postnatal ontogenesis; cranial venae cavae, ethanol.

Introduction

Morphology of major arterial and venous lines at different ages, and especially their changes under the experimental exposure gives a deeper understanding of problems in the structure of blood vessels in normal and pathological conditions.

Distortion of the structure of veins often leads to chronic venous insufficiency, varicose disease and thrombosis [1]. Effect of ethanol is primarily manifested in violation structures of the blood vessels and the heart [2, 3].

Ethanol has the ability to penetrate into the mammary gland and flow in milk [4]. In the available literature, we found no data on the effect of alcohol on the state and development of the vena cava in postnatal ontogenesis.

The purpose of the study. To study the changes of intrapericardial cranial venae cavae of rats in postnatal development under the influence of ethanol.

Materials and methods.

Investigated intrapericardial part of the right (RCVC) and left (LCVC) cranial venae cavae in 120 rats 6, 11, 16, and 22 to 30 days of ages. 60 of them received ethanol with mother’s milk. For this females were daily intragastrically administered 40 ° ethanol in a dose of 8 ml / kg during the lactation period (22 days).

The control group consisted of 60 rats of the same age, females – mothers who daily intragastrically received distilled water in the same dose.

The animals were kept under standard vivarium conditions at t 21-22 ° C and a natural photoperiod, on a normal diet. Slaughtering rats was conducted in accordance with the “Rules of work with experimental animals”, adopted by the Committee on Bioethics of the Republic of Uzbekistan.

Slaughtering rats was performed under ether anesthesia. After opening the thoracic and abdominal cavities material was fixed in 12% neutral formalin. Then vena cava was isolated and carried on the battery alcohols increasing concentrations, embedded in paraffin. Prepared histological specimens longitudinal and transverse sections of intrapericardial (orifice and non-orifice part – the area between the mouth and the attachment of the pericardium to the wall of the vein) part of the RCVC and LCVC.

Applied paint sections with hematoxylin and eosin, Van Gieson, Weigert methods, and impregnation method Foote under the modification N.A. Yurin. Using ocular line with magnification ob.90, ok.7.measured the thickness of the wall of the cranial venae cavae in the orifice and out parts. Mathematical processing of data was performed using the application Microsoft Excel 2010 in the section descriptive statistics.

The data obtained were processed statistically by standard methods for sampling with the definition of the mean (M), error of the arithmetic mean (m), quadratic deviation (σ), Student’s t test (t). Differences were considered significant at p≤ 0.05.

Results

In 6-day-old rats treated with ethanol through mother’s milk from the first day of birth, in the wall of the orifice of RCVC and LCVC subendothelial layer contains dense bundles of elastic fibers compared with the control. In the outer layer, elastic fibers in experimental rats are presented in separate fragments, or missing. In control rats the elastic fibers in the outer layer of the same area are absent. In non-orifice part of the RCVC and LCVC elastic fibers, located in the subendothelial layer, also have a greater density than the control. Reticular fibers form large incomplete loops.

At 11 days of age are most exposed to the effects of ethanol endothelial layer of the orifice part of the RCVC and LCVC. Endothelial cells contain pyknotic nuclei, which are located far from each other. At this age, the collagen and elastic fibers have a higher density than the control. Reticular fibers in experimental animals are placed chaotically in some places where they are totally absent. At 11 days of age, the walls of RCVC and LCVC non-orifice area in rats, received ethanol with milk, elastic fibers also thicker than in the controls. Given area in experimental rats in the wall of both cranial veins found zones devoid of reticular fibers. On day 16 of the experiment in comparison with the control, in the orifice part of RCVC and LCVC elastic and reticular fibers have different thickness and density of the location.

In some places, elastic and reticular fibers are thick, in others not. Collagen fibers are notably thickened and form dense tufts. In the mid – layer non – orifice part of the RCVC and LCVC, nuclei in myocytesare less than in controls. Collagen fibers of the RCVC under the influence of ethanol become much thicker than in controls. There is no significant change in the collagen fibers of LCVC at given age. Reticular fibers of the middle layer in many places are absent, and in those places where they are, have the form of non-closed loop. In 22-day-old rats exposed to ethanol, in the middle layer orifice part of the vein increase the area of non-nuclear sites in comparison with the 16-day-animals of given group. Elastic fibers become loose in some parts of the veins, in the other, on the contrary form dense tufts. In experimental rats, endothelial cells’ nuclei of the non-orifice part of the cranial venae cavae have elongated shapes, in some places desquamation of cells is observed. In the middle layer, the area of non-nuclear sites increases. Elastic fibers in the subendothelial layer are thinner than in the control group.

In the middle and outer layers, the elastic fibers are arranged as separate fragments. Reticular fibers of the RCVC and LCVC do not form loops and are scattered.

By 30-th day of the experiment the orifice part of the RCVC and LCVC is mainly covered by connective tissue. The outer layer is thicker than in the controls. Collagen fibers are densely forming the thick bundles, they occupy a large area of wall thickness. Elastic fibers between the subendothelial layer, and muscular tunic and at the boundary of the outer and middle layers loosened. On the 30th day in the non-orifice part, collagen fibers are thicker than in the orifice part, especially in RCVC. In carrying out morphometric studies revealed that the wall thickness of both paired cranial venae cavae in the orifice part is smaller than in controls. Differences were significant in RCVC and LCVC 11, 16, 22 and 30 days after birth (Table 1).

Increase in wall thickness of the orifice part RCVC and LCVC in 11-day-old rats compared with 6-day amounted to 10% and 8.3%. By 16 days of age, the thickness of the orifice RCVC and LCVC compared with the previous period increased by 24.9% and 16.1%. Increase in wall thickness of the RCVC and LCVC in the orifice part in 22 days rats decreased to 11.3% and 10% by day 30 it occurs differently in RCVC (17.8%) and LCVC (3.2%). Table 1

Wall thickness of the orifice part of RCVC and LCVC in experiment and control (micron), n = 50

age

(days)

RCVC

LCVC

experiment

control

experiment

control

6

16,1±0,45

17,4±0,51

20,6±0,52

21,5±0,65

11

17,7±0,47*

19,5±0,47

22,3±0,57*

26,5±0,69

16

22,1±0,47*

23,6±0,41

25,9±0,64*

28,7±0,82

22

24,6±0,63*

29,6±0,63

28,4±0,73*

33,0±0,67

30

29,0±0,65*

36,9±0,57

29,3±0,82*

38,1±0,56

         

* – Р<0,05 comparing both groups

In non-orifice area of 11-day-old rats in the experimental group, increase wall thickness of RCVC and LCVC as compared with 6 -day-old rats is slight – 7.2% and 5.1%. Wall thickness of 16-day-old experimental rats increased as compared with 11-day in RCVC to 21.6%, in LCVC to 17.1%.In 22-day-old rats increase of the wall thickness of RCVC is above and is 31.1%, whereas LCVC is 2.1%.The most substantial growth in thickness of the RCVC observed on day 30 of the experiment (56,3%), while of the LCVC, it was 19.3%. Consequently, the greatest increase in the thickness of the orifice part of RCVC and LCVC in experimental rats observed at 16 days of age. In non-orifice area under the influence of ethanol, a strong increase in wall thickness of the RCVC begins 16 days after birth, but it becomes a maximum on day 30, after the cessation of ethanol poisoning females. In LCVC growth occurs in waves, minimum on 11-and 22-days and increased on 16 and 30-days.

The wall thickness of both pair cranial venae cavae in non-orifice part in experimental rats on 6, 11, 16 and 22 days is less, and on day 30, it becomes larger than in control.

Certain differences in RCVC and LCVC on 11, 16, 22 and day 30(Table 2).

Table 2

Wall thickness of the RCVC and LCVC in non-orifice area in experiment and control (micron), n = 50

age

(days)

RCVC

LCVC

experiment

control

experiment

control

6

13,8±0,33

15,6±0,45

15,6±0,39

16,2±0,47

11

14,8±0,42*

17,2±0,48

16,4±0,55*

19,1±0,70

16

18,0±0,55*

20,5±0,45

19,2±0,58*

22,2±0,62

22

23,6±0,79*

25,9±0,80

19,6±0,49*

27,7±0,63

30

36,9±0,60*

34,6±0,83

23,4±0,64*

35,9±0,66

* – Р<0,05 comparing both groups

The study showed that in the context of alcohol intoxication, observed a breach of postnatal histogenesis of the intrapericardiac part of cranial venae cavae. Entering of ethanol for 6 days causes thickening of elastic fibers and decreasing reticular fibers in the non-orifice area. Prolonged alcohol intoxication leads to further destruction of reticular fibers and myocytes, degenerative changes of endothelial cells are occurred. Deformation of the vascular wall integrity, including tunica adventitia, found in alcoholic human disease in brain vessels.

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In the intima, also observed endothelial desquamation, loosening of muscular layer and adventitia [5]. 22 days after birth in experimental rats to marked changes in the wall of the cranial venae cavae joins thickening of collagen fibers. After the cessation of receipt of ethanol through mother’s milk (from 22 to 30 days) elastic and reticular fibers do not restore their structures, their loosening or formation of thick beams happens. There is a substantial thickening of the collagen fibers, which is a manifestation of multiple sclerosis of the vein wall. Destruction of elastic and reticular fibers of the vessels and the soft core of the spleen, the reduction of cell elements (smooth muscle cells, fibroblasts and fibrocytes) and connective tissue stromacollagenization reflect the aging process [6].

Wall thickness of the RCVC and LCVC in intrapericardial part throughout the experiment was significantly less, except RCVC on the 30th day in non-orifice area. In rats with alcohol intoxication disturbed growth rate of RCVC and LCVC. Previously we have determined that the wall thickness of RCVC a day after birth is 16,3 ± 0,44 micrometers and LCVC is – 17,8 ± 0,42 micrometers.

Substantial growth of wall thickness in the orifice part of RCVC occurs on 16-30 days, and LCVC – already on 6 and 11 days after birth, in non-orifice area of RCVC from 16 to 30 days, LCVC – from 11 to 30 days. In the

first 6 days under the influence of ethanol there is no increase in the thickness of the orifice part of RCVC and LCVC in 6-day-old rats in the experimental group compared with 1-day-old control group [7].

At a later date , increase the thickness of the orifice part of RCVC observed only on day 16,then the growth rate goes down, and in LCVC it occurs later than in the control, and also on 16th day. Increase in thickness of the non-orifice area of RCVC under the influence of ethanol does not significantly differ from control, while LCVC – significantly less in all periods of the study. Morphometric parameters of vessels reflect their functional features.

Pronounced morphometric changes indicative of impaired hemodynamic, had a vein of the liver lobules in 3 days after shin injury [8].

Thus, the effect of ethanol at the early stages of postnatal ontogenesis leads to pathological changes, which are expressed in endothelial desquamation, destruction of reticular, elastic fibers, myocytes and collagenization of connective tissue stroma. These changes occur with violation of histogenesis of the cranial vena cava wall that appears in the orifice and non-orifice areas of the intrapericardiac part. From both cranial venae cavae, the largest histogenesisviolation occurs in the LCVC.

References

  1. Shvalb P.G. , Ukhov Y.I., Tsaregorodcev A.A. Nature of the changes of the venous wall, depending on the cause of recurrent varicose veins. // Flebologiya. – 2009. – â„-4. p. 26-31.
  2. Porsukov E.A. On the morphological diagnosis of alcoholic heart lesion // Forensic Medicine. – 2009. – â„-6. – P.21-24.
  3. Steinweg D.L., Worth H. Alcoholism: the keys to the CAGE // Medicine.-1993.-Vol.94.-No.5.-P.520-523.
  4. Berejnoy R.V.,Smusin Y.S.,Tomilin V.V., ShirinskyP.P. Guidelines for forensic medicine poisoning. Moscow, M.1980, p.227.
  5. Krachun G.P., Pishenko E.E., Razygraeva N.L., Petrovina I.A., Kushnir P.X.Functional Histomorphology of intertissue changes in the wall of brain vessels, with alcoholism. // Xist, Chernov, issue 15. 2013, p.270.
  6. Alphonsova E.B., Functional morphology of connective tissue stroma of the spleen in the age aspect. // Achievements in Gerontology. Ed. “Aesculapius” (St. Petersburg). 2012-T 25, â„-3.- S. 415-421.
  7. Hadjiyeva A.U., Blinova S.A., Development of intrapericardial part of the cranial vena cava of rats in postnatal ontogenesis //Morphology.-2014, V.8, â„-1. P.26-29.
  8. Ocheretina R.Y., Mkrtchan O.Z., Strogov M.B., Morphometric parameters of vessels of the liver lobules in mice during the recovery period after shin injury.

 

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