Daskalov, D.

Gavazova, I.

Zaharieva, R.

English

Varna Medical University Press: Journals

COMPARATIVE INVESTIGATION OF T H E PRECIPITATING I N F L U E N C E OF HISTONES UPON PLASMA OF RATS W I T H TUMOURS AND H E A L T H Y ONES I . Gavazova , R . Z a h a r i e v a , D . Daskalov The histones (H) which are widely distributed in the tissues of mama" Hans, specially in connection wi th certain situations of cell destructions (tu­mour and necrotic cell damages, large traumas, granulocyte destruction, etc.), can enter the blood-stream (1) . Being already there they can form com-lexes wi th various plasma proteins and can undoubtfully influence on certain physiological functions (2, 3) . Our previous study shew that the plasma of healthy rats can be precipi­tated by H (4). Having in mind that the neoplastic processes are connected with an increased cell destruction and changes in the plasma we decided to investigate the precipitating effect of H on plasma of rats w i t h tumours. Materials and methods The study covers 27 rats wi th transplanted tumour of Walker . Para l le l ly wi th their plasma was tested the plasma of 25 healthy rats. A total H wi th in i t ia l concentration 300 micrograms was added to 0.5 ml plasma taken af­ter decapitation of each rat wi th E D T A . After 10 min at room temperature we read the precipi ta t ion,(+or—). I f there was no precipitation we increased the concentration of H unt i l obtaining i t . The plasma was analysed electro-phoretically in 17% P A A G , 2.5 M urea (5). H were obtained from rat l ivers by using 0.25 M H 2 S 0 4 (6) and diluted in destilled water. The protein was determined after the biuretic method (7). Results and discussion The added total H protein to rat plasma in vi t ro caused a precipitation in different degree of both normal plasma and that one from rats wi th tumours. The differences were considerable when small concentrations of H were used. As it can be seen from Table 1 wi th the results of our study, when H-concentra-tion was 300 micrograms 63% of the rats wi th tumours shew a ( + ) reaction, while for the control group the percent was only 25 .9%. W i t h increasing of Н-concen t ra t ion the difference between both groups (experimental and con­trol) was less and statist ically unre l iab le ; at a concentration 1000 micrograms the percent of rats from both groups wi th an established precipitation was equal. The electrophoretical analysis shows there are two ways of distribution of Н-f rac t ions in the supernatant of rats from both groups. The first one — Н-frac t ions H I and H2b-f-H2a are in the supernatant; second one — toge­ther wi th the cited H are found also the Н-fract ions r ich in arginin H 3 and H 4 . 22 I . Gavazova , R . Zaharieva, D . Daskalov T a b l e 1 P l a s m a precipi ta t ion at va r ious H-concentra t ions Amount oi added Н-protein in micrograms/0.5 ml Precipitation of rat plasma rats with tumours healthy rats - ' j + -300 500 1000 63 69 80 37 31 20 26 48 74 74 52 26 I n both cases together wi th a l l plasma fractions in the precipitate were detected H I , H 3 , H 2 b + H 2 a and H 4 . Our results show the different influence of H upon normal and patholo­g ica l plasma (rats w i t h tumours). Th i s is most wel l expressed when small concentrations of H were applied. The more H is added the bigger the per­cent of ( + ) reaction is and wi th 2000 micrograms the plasmas of a l l rats from both groups pract ical ly precipitate. We presume, that H , being strong base proteins, form complexes wi th plasma proteins. Changing of their isoelectri-cal point requires a possible forth-coming precipitation of the formed com­plex ( H + p l a s m a proteins). What is the reason of the various effect of H on normal and pathologi­cal plasma is s t i l l unknown. As for the neoplastic processes, perhaps there are certain changes of conformation, or also of aminoacid content of some plasma proteins. As a result of that they become more sensitive towards H-action and even small concentrations of a total H cause their precipitation. The various Н-f rac t ions have different effect on the plasma. Main ly the H r ich in arginin and less those r ich in lys in are these H which change the s tabi l i ty of plasma proteins and precipitate wi th them. I t is a probable result of the changed aminoacid content and conformation of H-fractions. The possible diagnostic value of the stronger effect of H on pathological plasma w i l l be the object of our future studies of patients wi th different di­seases. R E F E R E N C E S 1. Г у p о в и ч , А . Е . , С . И . Г о р о д е ц к и й , Е . В . С и д о р о в а . Био~ химия. 1967, 32 , 2, 3 0 2 — 3 0 9 . — 2. Д е л е к т о р с к а я , Л . И . , И . А . С е н т е б о * в а. Лаб. дело. 1971 , 8, 483—487 . — 3. D a s k a l o v , D . , I . G a v a z o v a . l Expc rientia, 34, 1978, 522 . — 4. L o s t i c k y , С , R . J e l i n e k . Physiol. Bohemoslov.> 25 , 1976, 341—344 . — 5. D a s k a l o v , D . , I . G a v a z o v a . I n : I n t e r n a t . Congr-P h y s i o l . S c i . , X X I I I . B u d a p e s t , 1980, A b s t r a c s , p. 370. — 6. P a n у i m , S . , R . C h a l k -ley. Arch. Biochem. Biophys., 130, 1969, 337. — 7. S p e l s b e r g , Т . C . , L . S . H n i -l i c a . Biochim. Biophys. Acta, 228 , 1971, 202. Comparative investigation of the. 23 С Р А В Н И Т Е Л Ь Н О Е И С С Л Е Д О В А Н И Е П Р Е Ц И П И Т И Р У Ю Щ Е Г О Д Е Й С Т В И Я ГИСТОНОВ НА П Л А З М У О П У Х О Л Е В Ы Х И З Д О Р О В Ы Х К Р Ы С И. Гавазова, Р. Захариева, Д. Даскалов Р Е З Ю М Е Д о б а в л е н и е т о т а л ь н о г о г и с т о н а в ы з ы в а е т н е о д и н а к о в у ю по с в о е й с т е п е н и п р е ц и п и ­т а ц и ю п л а з м ы о п у х о л е в ы х и з д о р о в ы х к р ы с . Р а з л и ч и я я р ч е в ы р а ж е н ы п р и б о л е е н и з к о й к о н ц е н т р а ц и и г и с т а м и н а . П р и д о б а в л е н и и 300 \ig в 6 5 % п л а з м ы о п у х о л е в ы х к р ы с п о л у ­ч а е т с я п р е ц и п и т а ц и я , в то в р е м я к а к в к о н т р о л ь н о й г р у п п е о н а д о с т и г а е т л и ш ь 2 5 % . П р и у в е л и ч е н и и к о н ц е н т р а ц и и г и с т о н о в о г о б е л к а эти р а з л и ч и я у м е н ь ш а ю т с я и п р и 1000 tig в ы р а в н и в а ю т с я . 

COMPARATIVE INVESTIGATION OF THE PRECIPITATING INFLUENCE OF HISTONES UPON PLASMA OF RATS WITH TUMOURS AND HEALTHY ONES

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COMPARATIVE INVESTIGATION OF THE PRECIPITATING INFLUENCE OF HISTONES UPON PLASMA OF RATS WITH TUMOURS AND HEALTHY ONES

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