Kankozha M.K.

Kazakh National Medical University of Asfendiyarov S.D., Almaty

STUDY OF THE ADSORPTIVE – TRANSPORT FUNCTION OF ERYTHROCYTES IN INTACT RATS AND THE RATS SUFFER FROM HYPOXIA

Hypoxia is a universal pathologic state, which appears with the most diverse states (conditions) of the human: with the respiratory and cardiovascular insufficiency, blood loss, and the disturbances of the cerebral and peripheral blood circulation [1]. Erythrocytopoiesis strengthens, the concentration of erythrocytes and hemoglobin increases, an oxygen capacity of the blood rises in response to the insufficient supply of organism by oxygen, and all of this is connected with the compensating ejection into the blood of endogenous stimulators of hematopoiesis – erythropoietins [2]. Adaptation to hypoxia can be in several directions. At the cellular level we increase anaerobic glycolysis due to an increase of the activity of the ferments which operate in glycolysis (phosphofructokinase, fructose-2,6- biphosphate and other).

From other side decreasing the process of energy intake due to the mechanisms which provide the income of oxygen into the organism, in spite of its deficiency in the environment (hyperventilation, the hyperfunction of myocardium, an increase in the oxygen capacity of blood) [3].

And finally, due to the processes which ensure a sufficient income of oxygen for vital important organs (increase of cell’s capability to utilize oxygen as a result of an increase of myoglobin concentration and lightening of the dissociation of oxyhemoglobin; the formation of new capillaries and an increase of permeability of cellular membranes) [4,5,6].

The purpose of this investigation was the study of the adsorptive- transport function of the erythrocytes of rats, which are located on the different stages of ontogenesis.

Materials and methods

Animals were divided into 2 groups: the 1st group – control group, and for animals of the 2nd group was invented the model of hypoxia.

Blood sampling conducted from the tail vein with the observance of the conditions of asepsis and antiseptics and also with the rules of humane relation to animals.

Blood was taken in centrifuged tubes with free flow mixing it with an anticoagulant (heparin) and slightly shaking (without foaming).

Blood was centrifuged (TLN-2) for 5 minutes at 1500 rpm in order to separate plasma and erythrocyte sediment from received blood. Then plasma was selected for consequent study, erythrocytic mass and 3% solution of sodium chloride were cautiously and thoroughly mixed in the ratio 1:3.

After that suspension was again centrifuged at 1500 rpm for 5 minutes. Then supernatant was picked and studied for a content of protein, albumen, cholesterol, triglycerides and alkaline phosphatase.

Indices led to the unit of erythrocytic mass: all obtained indices multiplied by 3. A quantity of adsorbed organic substances on the membrane expressed like concentration in plasma [7,8].

Static processing has been done on computer. The criteria of Styudent t - was significant at P <0.05.

Results and discussions

There is noted an increase of content of the glucose on the surface of erythrocytes in white pedigreeless rats, from immature animals to the mature. The level of glucose of the immature and mature animals suffer from hypoxia reliably exceeded control quantities 1,3 times (table 1).

A study of total protein in supernatant of the erythrocytes showed that in immature rats the level of protein under the conditions of hypoxia was reliably increased 1,2 in comparison with the intact. But in mature rats this index decreases 1,1.

Table 1. The level of glucose, total protein, albumen, cholesterol, triglycerides and alkaline phosphatase in supernatant of the erythrocytes of mature and immature rats within the standard and under the conditions of hypoxia (M±m).

Indices	Ontogenesis period of the white pedigreeless rats
	Immature		Mature
	Intact	Hypoxia	Intact	Hypoxia
Glucose (mmole/l)	2,73±0,03	3,54±0,03^*	3,06±0,03	4,14±0,09^*
Total protein (g/l)	18,3±0,27	21,48±0,30^*	28,53±0,30	26,25±0,33^*
Albumen (g/l)	5,91±0,12	8,19±0,27^*	9,42±0,36	12,42±0,48^*
Cholesterol (mmole/l)	1,05±0,03	0,48±0,01^*	1,38±0,03	1,92±0,02^*
Triglycerides (mmole/l)	0,69±0,01	0,96±0,02^*	0,84±0,02	0,54±0,02^*
Alkaline phosphatase (un/l)	43,23±0,09	39,45±0,06^*	42,36±0,06	45,51±0,06^*

Note:^* - Р≤0,05, in comparison with control.

The level of albumin in supernatant of the erythrocytes which obtained from rats of one-month increases in comparison with the control 1,4., than in rats of six months with hypoxia 1,3.

The content, obtained from the surface of erythrocytes, reliably decreased under the hypoxic influence in immature animals 2,2., in comparison with the control indices. But in mature animals the level of cholesterol reliably decrease 1,4.

Triglycerides in supernatant of the erythrocytes which obtained from rats of 1 month suffer from hypoxia reliably increase 1,4. On the contrary the observable indices of immature animals decreased 1,6 in comparison with the control.

The content of alkaline phosphatase in supernatant of the erythrocytes, isolated in the immature rats suffer from hypoxia reliably decreases; however, in mature rats reliably it grows 1,1, in comparison with the intact indices.

Thus, in the ontogenesis of white pedigreeless rats an increase of the level of observable indices in the supernatant of erythrocytes is noted.

In this case it is noted that the content of cholesterol and alkaline phosphatase in the supernatant decreases with hypoxia in the immature animals suffer from hypoxia, and the level of protein and triglycerides in the supernatant of erythrocytes decreases in the mature animals suffer from hypoxia.

The stability of organism to hypoxia is defined by the dependence of its metabolic processes from oxygen consist in environment, which was formed in the process of evolution and ontogenesis.

Literature:

1. Lukyanova L.D. Biochemical bases of the formation of mechanisms to the adaptation to hypoxia. M.Science, pages 161-164.

2. Sabanova R.K. Change of the mechanical resistance of the erythrocytes of animals which suffer from hypoxia. Successes of the contemporary natural science. 2006, №6, p. 45-46.

3. Hue L., Rider M.H. Role of fructose 2,6-bisphosphate in the control of glycolysis in mammalian tissues //Biochem J. – 1987. Vol. 245. Р.313–324.

4. Goranchuk V.V., Sapova N.I., Ivanov A.O. Hypoxytherapy. 2003, p.536

5. Gareev R.A. «The second function of erythrocytes». Collection of the scientific works of the international conference dedicated to the 150-anniversary of the institute of physiology of Pavlov I.P.- S-Petersburg. 1999 year, pages 25-27

6. International guiding principles for biomedical research involving animals. Geneva: Council for international organizations of medical sciences,1985. -P.187-195.

7. Zverkova E.E. «Blood supply of myocardium and resistance of organism to hypoxia with the trainings hypoxic-hypercapnic exposures». Alma-Ata, 1982.

8. Gareev R.A., Beklemishev I.B., Murzamadiyeva A.A. «Procedures of the study of hematolymphatic metabolism». Alma Ata, Science, 1991. p.135