Divanyan, H.

Kovachev, D.

English

Varna Medical University Press: Journals

PHYTOHEMAGGLUTININ E F F E C T ON T H E E L E T R O P H O R E T I C MOTILITY OF E R Y T R O C Y T E S D. Kovachev, H. Divanyan The electric charge of cells plays an important role in a l l physiologica processes. To secure normal fulfilment of their functions, the blood cells should possess a stable electric charge which is conditioned by the chemical structure of the cellular membrane, and by the composition of the surroun-ding mi l i eu . Th i s charge is altered in pathological conditions and as the result of external effects ( 1 , 6) . I n this regard, i t is of utmost importance to deter-mine the change in electrophoretic moti l i ty ( E P M ) of the cells caused by a variety of external factors, and in morbid conditions as w e l l . According to Roquet (5), there are phytohemagglutinin ( P H A ) recep-tor zones on the cellular surface whose number varies in the course of the su-bject 's individual development. Lesney (4) and Stuart (7) point out that the quantity of these zones on the erythrocyte membrane is different for P H A ob-tained from various vegetable species, wi th the quantity for P H A from Pha-seolus v i lgar is amounting to 520 000, for Robinia pseudoaccacia — 210 000, and for Lens culinaris — 5.5><105. P H A «Phaseolosaxin» is characterized by a high mitogenic and blast ac-t i v i t y , and marked proliferative processes' s t imulation (2). The preparation was obtained in the Chair of Biology at the Medical Facul ty — V a r n a , after the method of M . Tzoneva (3). I t is the purpose of the present work to inve-stigate the action of P H A phaseolosaxin on E P M of erythrocytes, in v i t ro . Material and methods Rabbit erythrocytes, undergoing three-fold washing, were used in the study. The erythrocyte mass obtained wes diluted wi th physiological saline at 1:10 ratio. Rabbi t red blood cells were used to render possible the compa-rative study of the results from planned future studies, in v i v o . Under expe-rimental conditions, the erythrocytes were diluted in physiological saline con-taining 20 y / m l P H A . The indicated P H A concentration exerts maximum m i -togenic effect on the blood cells, and i t is the most frequently employed in cytogenic researches. Red blood Cells, diluted wi th physiological saline were used as a control. To determine E P M of the erythrocytes, we measured the time required by the cell to move at a certrain distance within a microcham-ber, designed and constructed at the Chair of Biophysics, Biological Facul ty — Sofia Univers i ty . Measurements were carried out wi th ocular mesh and secun-dometer, observing sets of 50 cells each in microscope at one-fifth depth level of the chamber, at tension 180 volts and current force 4.5 and 6mil l iamperes. The different force employed contributes to the re l iab i l i ty of the results. The time required by erythrocytes to cover 1 mm distance — from right to left from left to right and vice versa — was measured three times. 40 Results and discussion Table 1 illustrates the results of the study performed. T a b l e t Group Exper. Control Exper. Control Exper. Control Pha effect on erythrocyte EPM Cur rent force Number of cells 4 4 5 5 6 б 50 50 50 50 50 50 8.77 5.70 6.80 4.09 3.78 3.80 8.77=1=0.34 5.70=1=0.19 6.80^=0.42 4.09-0.11 3.78=t0.12 3.80=!= 0.10 < 0.01 <0.01 >0.05 trol t The data from each experiment were compared separately with the con . Re l iab i l i ty of the results was calculated according to the var iat ional ana lysis rules. In the control setting, at current force 4 cement stance treated P < 0 . 0 1 I О/ \>Уд Control* j~~] Experiment mA, the mean time for displa-of erythrocytes at 1 mm di-is 5.70 sec, whils t in P H A erythrocytes — 8.77 sec, at . A t 5 mA, the time values .1. 4 5 6 mA Fig. 1. Correlation between time for ery­throcyte movement and electric current force. are accordingly 4.09 seconds for con­trols, and 6.80 seconds for the experi­mental groups. Only when 6 mA are used, there is no essential difference between control and experimental gro­ups. The above results are clearly demonstrated in Figure 1. The data obtained show that at force of the current 4 and 5 mA, the time required for cell movement in­creases as the result of electric po­tent ia l f a l l , whereas at 6 mA, the diference in velocity of movement of the erythrocytes between control and experimental groups disappears. The obser­ved reduction of the electric charge may be attributed to the fixation of P H A molecules on the surface of the cel ls . The'method applied did not allow for detailed elucidation of the mechanism of this particular phenomenon. I t is furthermore evident that changes in the potential under the influence of P H A might de recorded only upon application of a suitable electric f ield. The results presented clarify a single aspect of the biological act ion of P H A , and they should be beared in mind during its application, both in v i v o , and in vi t ro . Phytoheraagglutinin effect on the electrophoretic. 41 [ R E F E R E N C E S 1. Х а р а м о н е н к о , С , С. Р а к и т я н с к а я А. А. Электрофорез клеток в норме и патологии, «Беларусь», Минск, 1974, 41—94. — 2. Ц о н е в а , М. Хирургия, 1965, 6, 670—679. — 3. Ц о н е в а , М. Авторско свидетелство № 12008 от 20. I X . 1964— 4. L e s n e y , А. М. Arch. Biochetn. and Biophys., 1972, 153, 831—834. — 5. R о q u е t r. FEBS Abstracts, 1975, 110, 1000. — 6. S t о 1 t z, J . F . C. r. Soc. biol, 1970/71, 164, № 6, 1321 — 1324. — 7. St и ar t, K. J. Biol. Client., 1971, 246, № 21, 6581—6586. В Л И Я Н И Е Ф И T О Г Е МО Г Л О Б У Л И Н А НА ЕЛ Е К Г Р О Ф О Р Е Т ИЧ Е( К УЮ П О Д В И Ж Н О С Т Ь Э Р И Т Р О Ц И Т О В Д. Ковачев, X. Дизанян Р Е З Ю М Е Определена электрофоретическая подвижность эритроцитов зайца в микрокамере . Прослежено время перемещения на оиределеное расстояние . Наблюдается снижение электрического заряда красных кровяных клеток под действием 20у/мл Ф Х А . Допускается , что Ф Х А фиксируясь на клеточ­ной мембране ведет к изменению электрического з а р я д а . Эти изменения наблюдались только в соответствующем электрическом поле. 

PHYTOHEMAGGLUTININ EFFECT ON THE ELETROPHORETIC MOTILITY OF ERYTROCYTES

https://journals.mu-varna.bg/index.php/ssm/article/download/3452/2911

PHYTOHEMAGGLUTININ EFFECT ON THE ELETROPHORETIC MOTILITY OF ERYTROCYTES

Abstract

Similar works

Full text

Available Versions

Varna Medical University Press: Journals