The human pathological roles op the superoxide radical and superoxide dismutase

A C T A U N I V E R S I T A T I S L O D Z I E N S I S FOLIA BIOCHIMICA ET BIOHiYSICA 2 , 1983

B e l a M a tko vio s

THE HUMAN PATHOLOGICAL ROLES OP THE SUPEROXIDE RADICAL ( 0 ~ ) AND SUPEROXIDE DISMUTASE (SO D )*

The p r e s e n t l e c t u r e i s i n t e n d e d to o u t l i n e th e r e s u l t s d e m o n s t r a t i n g th e r o l e s o f 0 2 and SOD i n human d i s e a s e s . We s h a l l a t t e m p t to b r i n g th e l i t e r a t u r e d a t a i n t o c o r r e l a

-t i o n w i -th our own r e s u l t s o r w it h our e x p e r i m e n t a l c on cep -t i o n s .

The human r o l e o f th e 0~ r a d i c a l w i l l be d e a l t w it h , f i r s t , f o l l o w e d by th e SOD ch a n g es and t h e i r c o r r e l a t i o n s .

The r o l e o f th e 0 " r a d i c a l h e r e i s n ot e x c l u s i v e . The v a r i o u s r a d i c a l s formed from m o l e c u la r oxyg en may i n t e r a c t w ith one a n o t h e r , r e s u l t i n g i n f u r t h e r a c t i v e r a d i c a l s su ch a s the h y d r o-x y l r a d i o a l (O H *), s i n g l e t oo-xygen ( 10 2 ) , peroxy and h yd rop eroxy r a d i c a l s . The m u l t i d i r e c t i o n a l r e a c t i o n pathway t h a t r e s u l t s i n a c t i v e oxygen r a d i c a l s i s summarized i n d e t a i l i n th e t a b l e s to be fou nd i n th e p u b l i c a t i o n by S i n g h [1 8 ]. T h i s c o m p i la t i o n was p re p a re d f o r th e symposium d e a l i n g w ith oxy gen r a d i c a l s , o r -g a n i z e d i n Pinawa , M anitoba i n 1977 by W h i t e s h e l l N u c l e a r Re-s e a r c h E Re-s t a b l i Re-s h m e n t Atomic Energy o f Canada, L t d . The p r o c e e d in g Re-s o f th e symposium were p u b l i s h e d i n one oopy o f 28 th e volume o f th e Photoohem. P h o t o b i o l . The H a b e r-W e i s s and F e n -t o n r e a c t i o n [ 7 ] a r e w e l l known s o u r c e s o f OH* r a d i c a l s :

* L e o t u r e d e l i v e r e d on th e P o li s h - H u n g a r ia n Syp osiu m "The e f f e c t o f p h y s i c a l and c h em ioa l a g e n t s on m a c r o m o l e c u l e s " , Łódź ( P o l a n d ) i 1 8 - 2 0 th o f Ju n e , 19 79 .

0 2 + OH" + OH* ( a )

and

L e t ua o o n a i d e r f i r a t the e f f e c t o f 0 " r a d i o a l a on r e d b l o o d o e l l a (RBC). The damage oa u aed to th e f o r m a l e le m e n ta o f the b lo o d may be d i r e c t or i n d i r e c t . In b o t h c a a e a the o v e r a l l r e a u l t i a th e aame: h em o l y a ia namely l y a i s o f th e RBC membrane. T hia e f f e c t , i n which th e 0” r a d i c a l p la y e an i m p o r t a n t r o l e , haa b een aummarized i n a T a b . 1 [ 4 ] .

T a b l e 1

0 " r a d i c a l a a p o t e n t i a l c a u a e a o f o x i d a t i v e h e m o ly a ia ( d e t a i l s i n th e t e x t )

R od nik Ol ja k o p o t e n c j a l n a p r z y c z y n a hem oliz y oksyd a tyw n ej ( a z c z e g ó ły w t e k ś c i e )

0~ pajtHKaji kbk noTeHUHajiBHafl npmiHHa OKCHflaTHBHoro re»iojiH3a

2 (nOflpoOHOCTH B T6KC T6 ) I n c r e a s e d f o r m a t i o n o f Og D e o r e a ae d p r o t e c t i o n a g a i n a t them 1. V ia hb. 3 . D e f i c i e n c i e s o f : U n s t a b l e h b a. G l u c o a e - 6 -p h o a p h a t e d e h y d ro g e n a se T h a l a s s a e m i a 6 -P h o ap h o g lu c o n a t e d eh y d r og en a ae O x i d a t i v e d ru g s G l u t a t h i o n e p e r o x id a s e (GPx) e . g . A c e t y l p h en y l h y - G l u t a t h i o n e r e d u c t a s e d r a z i n e G l u t a t h i o n e S y n t h e t a s e . A n t i m a l a r i a l s C a t a l a a e F av ism SOD PQ ( p a r a q u a t ) V i t . E H eat M e ta l i o n s (Cu2 + , Fe 2+ e t c . ) Redox a u b a t a n c e s 2 . D i r e c t e f f e c t a R a d i a t i o n H y p e r b a r ic oxygen P o rp h y r i a ( f r e e p o r-p h y r in a )

The mechaniam i s d e a l t w i th i n two f i g u r e s . F i g u r e 1 o u t l i n e s th e hem p ocke t o f th e oxyhem og lobin (oxy hb ) m o l e c u l e . Sm a l l

c h a n g es may l e a d q u io k l y t o th e f o r m a ti o n o f methemoglobin (m e th b ). Suoh a ohan ge may be c a u s e d by th e w a te r m o le c u le or s m a l l a n io n s; t h e s e d on a te an e x t r a e l e o t r o n to m o l e c u l a r to m o l e c u la r oxygen c o n v e r t i n g t h i s t o th e 0« r a d i c a l , Fe2 + i s

o ,

d.

o x i d i z e d to Fe , and methb i s formed from oxyhb .

F i g u r e 2 sum m a riz es the meohanisms which p r o t e c t th e RBC from h e m o l y s i s .

T og ethe r w ith T a b . 1 F i g . 2 s h e d s more l i g h t on the d e t a i l s o f e ac h meohanism.

R educed g l u t a t h i o n e (GSH) and oxyhb form th e f i r s t l i n e o f d e f e n c e o f the RBC a g a i n s t o x i d a t i v e damage. The se c o n d p r o t e c -t i v e l i n e i n c l u d e s -SH m a -t e r i a l and p o l y u n s a t u r a t e d f a t t y a c i d s (FUFA ). F i n a l l y , o x i d a t i o n o f t h e HJF.A by l i p i d p eroxid ation ( L P ) , l e a d s t o i r r e v e r s i b l e damage o f t h e RBC membranes and to h e m o l y s i s .

One o f the most i m p o r t a n t o f th e ab ove f a c t o r s l e a d s to h e-m o l y s i s d i r e c t l y , and the o th er one i n d i r e o t l y ( s e e T ab . 1 ) . The same r e f e r s t o th e i n h e r i t e d f a c t o r s t o o .

A

d i r e c t hb e f f e c t i s i n v o l v e d i n t h a l a s s a e m i a , f o r e xa m p le, w h i le a d e c r e a s e o f th e d e f e n c e a g a i n s t th e 0“ r a d i c a l r e s u l t s i n d i r e c t l y from th e v a r i o u s i n h e r i t e d enzyme d e f e c t s , ma inly th e d e o r e a s e o f th e SOD s y n t h e s i s , or a c a t a l a s e m i a , f o r i n s t a n c e . We s h a l l n ot d e a l h e re w it h h e m o l y s e s c a u s e d by d i f f e r e n t r a d i a t i o n e f f e c t .

Mention may now be made o f d i s e a s e s a f f e c t i n g one or g a n o r th e e n t i r e o r g a n is m , i n which an i n c r e a s e i n th e q u a n ti t y o f th e 0“ r a d i c a l p l a y s an i m p o r t a n t r o l e .

P r i m a r i l y from the work o f M o C o r d [11 ] i t i s known t h a t i n v i t r o th e

0^

r a d i c a l s p o s s e s s the a b i l i t y to d e p o l y - m e r i z e h y a l u r o n i c a c i d . D i r e c t c o n s e q u e n c es o f t h i s p u b l i c a -t i o n were -t h o s e i n v e s -t i g a -t i o n s t h a t a tte m p te d t o c l a r i f y the r o l e s o f

0~

and SOD i n r h e u m a tic d i s e a s e s . U n f o r t u n a t e l y , a u n if o r m p i o t u r e h a s n ot y e t d e v e lo p e d i n t h i s f i e l d but i n our view i t i s worth the t r o u b l e . By the h y p o t h e s i s o f MoCord and c owo rkers 0“ formed i n g r e a t e r amount, or p o s s i b l y a d e-c r e a s e d SOD a e-c t i v i t y , l e a d s to th e f i r s t a t t a e-c h whie-ch d e v e l o p on th e commencement o f rh e u m a t ic d i s e a s e s , t h i s i s one p a r t o f our working h y p o t h e s i s t o o . F u r th e r q u i t e s i m i l a r l y t o a u t o c a t a - l y t i c p r o c e s s e s , i n tim e th e o t h e r f a c t o r s d e s t r o y themselves ; m ai n ly th e r e c u r r i n g i n f l a m m a t i o n s , and then l a t e r th e d e s t r u c

-Schemat n i e k t ó r y c h r e a k c j i w k i e s z e n i hemowej hemoglob iny C x e n a H e K 0 T 0 p u x p e a K u H i ł b r e u 0 B 0 M K a p M a H e r e M o r j jo O H H a

N A D P H N A D P 2 . P ro d u c ti o n an d r e d u c ti o n o f a c ti v a te d o x y g e n r a d ic a ls a n d th e ir c o n n e c ti o n s to th e g ly c o ly ti c an d p e n to se p h o sp h a te p a th w a y s. T h e a c ti o n o f h e m o ly ti c fa c t o r s li s t e d in T ab . 1 (n u m b e rs in d ic a te d ). G P X -g lu ta th io n p e r o x id a se ! P P S -p e n to se p h o sp h a te sh u n t P o w st a w a n ie i r o z k ła d ak ty w n y ch ro d n ik ó w tl e n o w y c h i ic h z w ią z e k z g li k o li z ą i cy k le m p e n to z o - fo sf o ra n o w y m . W p ły w c z y n n ik ó w h e m o li ty c z n y c h p rz e d st a w io n o w ta b . 1 (z a c h o w a n o n u m e ra c ję ). G P x -p e ro k sy d a z a g lu ta ti o n o w a ; P P S -c y k l p e n to z o fo sf o ra n o w y 0 6 p a 3 O B a H H e h p a c n a . n a x T H B H u x k h c j i o p o a h u x p a ą m c a j i o B h h x c B H 3 b c t j i h k o j i h s o m h n e H T 0 3 o $ o c $ a T K U M U H k j i o m . E s H H H H e r e M O J Z H T H M e c K H x $ a K i o p o B n e p e ^ H C J i e H O b i a 6 . 1 ( R O M e p a y n a s a i c j ) . G P X -r j i y r a -T H 0 H -n e p 0 K C H f l a 3 a ; P P S -n e H T 0 3 C K ï > 0 c < i > a T H H f t u h k j i

t i v e f a c t o r s a r e c o n n e c te d w it h autoimmune p r o o e s a e s . S c u d- d e r e t a l . [17] compared the SOD a c t i v i t i e s o f RBC from 50 norm al and 50 rh eu m a t oid a r t h r i t i a i n d i v i d u a l a . No e s s e n t i a l d i -f -f e r e n c e i n SOD v a l u e s waa -found between th e two g r o u p s , how-e v how-e r , th how-e RBCCu c o n thow-e n t i n th e f e m a le rh e u m a to i d a r t h r i t i c s was s i g n i f i c a n t l y lower th an i n th e normal i n d i v i d u a l s . So f a r , t h e r e f o r e , a c o r r e l a t i o n h a s n ot been d e m o n s tr a t e d between th e RBC SOD a c t i v i t y d e t e r m i n a t i o n s i n a c t i v e rh eu -m a to id a r t h r i t i c s and th e i n v i t r o e x a m i n a ti o n s and th e or y o f M c C o r d . C o n ti n u in g our workin g h y p o t h e s i s i n i t s e n t i r e t y i s a s f o l l o w s : the p h a g o o y ta b u r s t i n g o f the p oly m or ph on u clea r l e u k o c y t e s (PMNL) and h en ce t h a t o f th e m a c rop h ag es, r e s u l t h ig h amount oxygen r a d i c a l s . The oxyg en r a d i c a l s , and mainly the 0“ a n i o n and th e H20 2 , er od e th e a r t i o u l a r s u r f a c e , s i n c e they d e p o ly m e r iz e th e h y a l u r o n i c a c i d [ 1 1 ] and th e ^2 ^2 g i n g th e SOD p r o t e i n [ 3 ] . The worned a r t i o u l a r s u r f a c e enhan- oea th e i n f l a m m a t i o n , which l e a d s to f u r t h e r PMNLs m i g r a t i n g i n . A v i c i o u s c i r o l e th e reb y a r i s e s , which i s b roken only by t h e r a p e u t i c i n t e r v e n t i o n , f o r example l o c a l SOD [ 8 ] s t e r o i d t r e a t m e n t e t c .

We w is h e d d e m o n s tr a te th e s i g n s o f the r h e u m a t i c d i s e a s e g r o u p i n th e a r t i c u l a r f l u i d . C l e a r l y , t h i s c o u ld be examined only i n a c u t e i n f l a m m a t i o n s , f o r only s t e r i l e w ash ing f l u i d may be o b t a i n e d from the i n f l a m m a t i o n - f r e e j o i n t , t h i a f l u i d h a v i n g a c h a r a c t e r i s t i c c o m p o s i ti o n . A few d a t a a r e p r e a e n te d h e re t h a t can be b r o u g h t i n t o c o r r e l a t i o n w i th our own e x a m i n a t i o n s . T a b l e 2 Normal v a l u e s f o r s y n o v i a l f l u i d W ar to śc i normalne d l a p łynu maziowego H o pM ajib H u e 3 H a M eH aa a j i h cHH OBH ajibHO# «h^k o c t h

C o n s t i t u e n t s A verag e Range T o t a l a lb u m in and g l o b u l i n 1 .7 2 g / 1 0 0 cm3 1 . 0 7 - 2 .1 3 g / 1 0 0 cm3 U r ic a c i d 3 . 6 mg/100 cm3 3 . 3 - 4 . 7 mg /1 00 cm3 L e u ko c y te count 63/mm3 13-180/mm3 B a s e d o n : H o l l a n d e r I . L . ( E d . ) , A r t h r i t i s and a l l i e d c o n d i t i o n s , P h i l a d e l p h i a , PA ( 1 9 6 0 ) .

Our e x p l o r a t o r y s t u d i e s con nec ted w ith a r t i c u l a r f l u i d were r e s t r i c t e d t o th e p e r o x i d e m etab olism enzymes (PME) (SOD, P - a s e and C - a s e ) , a c i d p h o s p h a t a s e (A P h -a se ), u r i c a c i d , p r o t e i n and Cu d e te r m i n a t i o n s .

The exa m in a ti on were p erform ed w ith Dr. I . A. K o rd o ss . On th e b a s i s o f o u r r e s u l t s , th e examined c a s e s were d i f f e r e n t i a t e d i n t o g r o u p s a c c o r d i n g t o th e d e g r ee o f s e v e r i t y a s i n d i c a t e d by c e r t a i n c y t o l o g i c a l and im m unolog ical d e t e c t i o n s .

I t i s c l e a r from th e Tab. 3 / P a r t B t h a t th e PME behave d i f -f e r e n t l y i n t h e -f o l l o w i n g g r o u p s . In g r o u p s I-V I t h e r e i s a lw ays gamma l a t e x p o s i t i v i t y . T h is means t h a t th e a r t i c u l a r f l u i d con-t a i n s a n con-t i b o d i e s con-t h a con-t r e a c con-t w icon-th con-th e ancon-ti-immune serum. As a r e a g e n t s u i t a b l e f o r s l i d e a g g l u t i n a t i o n ( l a t e x f i x a t i o n t e s t ) , Gamma-latex i s marked by Human ( B u d a p e s t ) . The r e a g e n t i s a p p l i -c a b l e f o r d e m o n st ra t io n o f th e rheu matoid f a c t o r (IgM ). The o t h e r q u a l i t a t i v e r a p i d t e s t t h a t we us ed was th e LE-TEST1^ (Hyland , USA), which can be employed f o r th e d e t e c t i o n o f a n t i n u c l e o p r o - t e i n f a c t o r s . L E - T E S T ^ - p o s it i v i t y o c c u r r e d only once o r tw i c e in a l l th e examined c a s e s .

T a b l e 3

Enz ymatic a c t i v i t i e s in s y n o v i a l f l u i d Aktywności enzymatyczne w p ł y n i e maziowym 3 H 3 H M a T t m e C K H e a K T H B H O C T H B C H H O BH aJIb H O tt K H Ą K O C TH

P a r t A

G e n era l

n o t a t i o n s None Verylow Low Normal

Modera-t e l y h ig h High Extremly h ig h V a lu es 0 - - _{N + ++ +++} SOD U/ml s y n o v ia l f l u i d ( s . f . ) - 10 N 10-15 + 1 5-2 0 ++ 20 P - a se U/ml s . f . 0 - 1 + 1-2 ++ 2 -1 0 +++ 10 C - a se BU/ml s . f . - 0 . 1 0 N 0 . 0 1 0

P a r t B Group I I I I I I IV V VI VII V I II SOD U/ml N N ++ + N N ++ N P - a s e U/ml +++ ++ + 0 - ++ 0 ++ Cu g/ml - - - - N — — -P r o t e in mg/ml - N N - - N N -C - a se BU/ml N + N - N - N N U ric a c i d mg/100 ml - - N N - ++ N N APh-ase U/ml +++ +++ ++ ++ ++ ++ + N R hagocyta + + - + - + - - - - + Gamma-latex + + + + + - - -APh-ase = a c i d p h o s p h a t a s e .

N o t e : The f i r s t p a r t o f th e Tab. 3 / P a r t A shows th e n o ta -t i o n s u s e d i n -th e q u a n -t i -t a -t i v e e v a l u a -t i o n o f -th e mea surements, and th e r a n g e s o f th e a c t i v i t y u n i t s / m l s y n o v i a l f l u i d f o r th e i n d i v i d u a l enzymes ( s e e Tab. 3 / P a r t A).

The sec ond p a r t o f th e Tab. 3 ( s e e P a r t B ) c o n t a i n s th e sum-mary, a b o u t which a few words w i l l be s a i d . I t may be se en a t a g la n c e t h a t th e most l e f t - h a n d column o f t h e P a r t B shows th e most p o s i t i v e s i g n s ; on p ro c e e d i n g from l e f t t o r i g h t t h e number o f p o s i t i v e s i g n s d e c r e a s e s ( a s t h e s e r i a l number i n c r e a s e s ) . On th e b a s i s o f th e ga mm a-latex r e a c t i o n , columns I-V mean f r e s h in f l a m m a t io n , which may be a f i r s t in f l a m m a t io n , b ut most o f te n i s an a c u t e r e c u r r e n c e . Group VI i s t h e g rou p o f p a t i e n t s w ith g o u t , o r a s t r o n g ten denc y t o t h i s .

With th e c o l l a b o r a t i o n o f Dr. J . J . Arvay, they r e p e a t e d th e ex a m in a ti o n s o f S c u d d e r e t a l . [ 1 7 ] on a c t i v e rh eu m atic p a t i e n t s a t th e N a t i o n a l Rheumatology and B a ln e oth er a p y I n s t i -t u -t e ( B u d a p e s -t ) . The r e s u l t s a r e p r e s e n t e d i n th e f o ll o w in g Tab. 4.

Some c hose n p h y s ic o -c h e m i c a l p r o p e r t i e s o f b l o od from p a t i e n t s w ith a r t h r i t i s ( g o u t ) and c o n t r o l group

Wybrane w ła ś c i w o ś c i fi z y ko c h em ic z n e krwi u p a c jen tów z a rtretyzm em i u osób ko n tro ln y c h

HeKOTopNe cJ)H

3HK0xHMJmecKHe cB o iicT B a kp o b h y CojibHu x ap ipH T OM

u y KOHTpojibHoił r p y n r w P a r t A C o n tro l Rheumatoid a r t h r i t i c p a t i e n t P a ra m ete r s n » 10 n A= 4

.

n B= 20 n C= 16 a ve -r a g e ra ng e a ve-r a g e rang e a ve -r a g e ra nge a v e -r a g e rang e SOD U/g hb 623 4 7 4-9 29 683 56 2-90 4 338 45 -4 7 4 642 4 94 -8 50 Sed im en ta-t i o n mm/h 14 6 -2 0 17 15 -2 0 45 2 0 -68 45 23 -10 4 RBC i r o n p q /1 0 0 ml 70 6 0-1 0 5 70 42 -1 43 74 22-122 53 14-121 RBC c o pp e r (jq/10 0 ml 119 67-151 144 9 5-17 5 150 99 -18 8 152 58 -2 1 9 P a r t B C o n tro l R heumatoid a r t h r i t i c p a t i e n t P a r a -m et ers n = 10 _n A = 5 n B = 8 n C = 23 n D = 4 a v e -r a g e ra nge a v e - r a ge ra nge a v e -r a g e ra nge a v e -r a g e ra nge a v e -ra ge ra n g e C - a s e U / g hb 4 . 3 2 . 6 - 5 . 8 4 . 3 3 . 9 - 4 . 6 2 . 1 1 . 7 - 2 . 5 4 . 0 2 . 8 - 5 . ? 7 . 3 6 - 9 . 2 Sedim en-t a en-t i o n mm/h 14 6 - 2 0 18 1 5 - 2 0 43 3 1 - 1 0 4 4-3 2 1 - 7 0 5 6 2 4 - 6 8 RBC i ro n uq/ 1 0 0 ml 70 6 0 - 1 0 5 61 2 2 - 1 4 3 4 0 2 0 - 7 7 75 1 4 - 1 2 4 65 2 0 - 1 0 4 RBC copp er j i q / 100 ml 119 6 7 - 1 6 1 14 2 9 5 - 1 7 5 1 66 1 1 5 - 2 0 0 143 5 8 - 2 1 8 172 13 9 -2 19 N o t e : I t can r e a d i l y be se e n t h a t , w ith a p p r o p r i a t e c l a s s i f i c a -t i o n , d i f f e r e n c e s ca n be d e t e c t e d a s r e g a r d s t h e RBC SOD, C - a s e and a l s o Cu c o n te n t i n th e a c t i v e , bu t t r e a t e d r h e u m a t i c s .

Of c o u r s e , th e d e p r e s s e d SOD l e v e l d o e s n o t a lw a y s mean a rh eum a tic d i s e a s e , b u t i n g e n e r a l i t may be a symptom o r a consequ enc e o f some a c u te o r c h ro n i c s y st e m i c d i s e a s e . T hese d i s e a -s e t y p e -s a l -s o l e a v e t h e i r mark in th e m etab ol i-sm o f th e RBC (more s im p ly , they a r e accompanied by a change i n SOD a c t i v i t y , which u s u a l l y means an SOD d e c r e a s e ).

We ha ve t h e r e f o r e g iv e n a working h y p o t h e s i s i n t h i s c a s e , and s u p p o rt e d t h i s w ith d a t a g r ou p s in th e f i e l d o f a r t i c u l a r s y st e m ic d i s e a s e s . N a t u r a l l y , t h i s d oe s n o t y e t mean th e p r o o f o f t h e working h y p o t h e 'si s. In ou r view , many fu nd a m enta l exa-m in a ti o n s w i l l be n e c e s s a r y i n t h i s f i e l d , in th e i n t e r e s t o f c o n f i r m a t i o n from v a r i o u s a s p e c t s .

Only mention w i l l be made o f a n o t h e r t o p i c , th e human aspects o f which we have i n v e s t i g a t e d . T hese e x a m in a t io n s were p erf orm ed on a n im a l s w ith d i a b e t e s in d u ce d e x p e r im e n t a l l y w ith ch em ic a l a g e n ts ( n a t u r a l l y , h e re t o o we can not g i v e an a ss u m p tio n pro ved i n every r e s p e c t from an e x p e r im e n t a l p o i n t o f view ; a t th e mo-ment I ca n on ly r e p o r t d a t a which f i t i n w it h ou r h y p o t h e s i s ) .

In d i a b e t e s , which i s wellknown s i m i l a r l y t o become a d i -s e a -s e a f f e c t i n g t h e e n t i r e or g a n i-sm , t h e o x i d a t i v e damag es w i l l be g r e a t e r th a n in o t h e r d i s e a s e s . B e c a u se o f t h e s e enhanced o x i d a t i v e damag es, a l l t h o s e symptoms s u b s e q u e n t ly d e ve lo p which a r e m a n i f e s t e d in v a s c u l a r s c l e r o s i s , n e u r i t i s and c a t a r a c t . In th e m ed ic a l te r m in o l o g y , t h e s e a r e th e se co n d a ry symptoms o f d i a b e t e s . F o r th e p a t i e n t , however, t h e s e c h an g es to o a r e p r i -mary and a r e ve ry f r e q u e n t l y th e c a u s e s o f d e a t h .

I sh o u ld l i k e to s t a t e h e r e one o f ou r fund am ental concep -t i o n s i n c o n n e c -t io n w i -th d i a b e -t e s de ve lop men -t, and w ith v e r t e -b r a t e m et a-b o lis m t y p e s i n g e n e r a l . A cc ord in g t o t h i s , humans and g i v e n s p e c i e s o f v e r t e b r a t e s in g e n e r a l can be d i v i d e d i n t o two o r more g r o u p s a s r e g a r d s th e ty p e o f th e o x i d a t i v e m eta b o lism . S i g n i f i c a n t d i f f e r e n c e s can be d em on str at ed in th e d i f f e r e n t g r ou p s in th e a c t i v i t i e s o f th e o x i d a t i v e m eta b olism enzymes. Hence, i n a f u r t h e r c o n c e p ti o n , d i a b e t e s d e v e l o p s p r i m a r i l y i n

i n d i v i d u a l s i n h e r i t i n g a h ig h e r o x i d a t i v e m eta b o lim s. S om a t o st a -t i n and i n s u l i n , f o r exam ple, a r e w e l l known t o be o x i d i z a b l e s u b s t a n c e s .

L e t u s c o n s i d e r f i r s t o u r own human d a t a r e l a t i n g t o SOD and in g e n e r a l th e o t h e r PME. T hese measurements r e f e r to t r e a t e d

d i a b e t i c s o f v a r i o u s o r i g i n s . The d a t a a p p l y only t o human RBC (T a b . 5 ) .

T a b l e 5

Some chosen p a r a m e t e r s o f b l o o d from d i a b e t i c s p a t i e n t s and c o n t r o l -group

Wybrane p a ra m etry krwi osób k on tro ln y c h i d ia betyków H e K0T0p u e n a p a M e T p n k p o b h K O H T p o j i b H u x 4 0H0p0B H HHaOeTHKOB P a ra m eter s C o n tr o l s D i a b e t i c s G lu c ose 8 1 . 5 2 1 0 .3 mg/100 ml b lo o d + 1 0 .4 + 5 4 .7 P r o t e i n 8 9 .4 7 8 .3 mg/ml p las m a + 4 .6 +1 5.1 P r o t e i n 5 8 9 .8 5 0 1 .2 mg/ml h e m o l y s a te s + 1 3 1 .5 + 1 7 2 .9 G3H-P0D 6 . 4 3 1 6 .4 5 U/ml h e m o ly s a t es + 0 .6 2 + 0 .5 4 C - a s e 0 .4 0 0 0 .0 2 7 BU/ml p lasm a + 0.1 41 + 0.0 01 C - a s e 0 .4 5 8 2 . 1 5 BU/ml h e m o l y sa t e s + 0 .0 5 5 + 0 .0 8 SOD 2 0 3 .5 3 . 0 U/ml h em o ly s a te s + 4 0 .3 + 0 . 3 .

N o t e : The Tab . 5 shows th e most im p o rt an t p a r a m e te r s and enz ym atic a c t i v i t i e s measured by hu-man b lo o d s a m p le s . The C - a se v a l u e s a r e e x p r e s s e d i n Bergmeyer U n it s (B U) ( n = 1 00 , X + S ) .

The f o l l o w i n g t a b l e p r e s e n t s s i m i l a r enzyme a c t i v i t i e s meas-u r e d in r a t s w ith e x p e r i m e n t a l l y -i n d meas-u c e d d i a b e t e s (T a b . 6 ) .

in t h e an im al e x p e ri m e n ts , o f c o u r s e , t h e r e was a l s o a p o s -s i b i l i t y f o r co mp a ris on o f th e (normal and d i a b e t i c ) v a l u e s o f th e enzyme a c t i v i t i e s o f th e d i f f e r e n t o r g a n s .

When t h e s e d a t a a r e examined t o g e t h e r , th e d i f f e r e n c e s a re s t r i k i n g : 1) in human b lo o d th e p r o t e i n v a l u e s lo w e r, 2 ) th e g l u t a t h i o n e p e r o x i d a s e a c t i v i t y i s a b o ut 3 t i m e s h i g h e r than in th e normal c a s e s , 3 ) th e c a t a l a s e a c t i v i t y o f th e RBC i s a bout 5 t i m e s t h a t o f th e norm al, 4 ) RBC SOD a c t i v i t y i s very low.

T h ese f a c t s p o i n t t o an enhanced H202 p ro d u c ti o n and decom-p o s i t i o n , and a d e c r e a s e d 02 r e s i s t a n c e .

The d a t a from th e a n im al e x p e rim e n ts s u p p o r t t h e s e same o b s e r -v a t i o n s [ 1 0 ] . T here i s o t h e r l i t e r a t u r e ev id e n ce i n s u p p o r t o f ou r hypo-t h e s i s : 1. I t i s known t h a t th e c a u s e o f th e r a p i d o c c l u s i o n o f th e d u c tu s a r t e r i o s u s and u m b i l i c a l a r t e r y a f t e r b i r t h i s t h e e l e -v a t e d 02 t e n s i o n (C>2 t o x i c i t y ) and d e c r e a s e d SOD a c t i v i t y o f t h e s e a r t e r i e s [ 6 ] . T a b l e 6 The i n f l u e n c e o f c h e m i c a l ly - in d u c e d d i a b e t e s on t h e a c t i v i t y o f SOD, P - a s e , C - a s e v a l u e s f o r r a t s

Wpływ cu krz yc y ind ukowanej c h e m icz n ie na a ktyw nośc i SOD, p er ok sy d a z y i k a t a l a z y u szcz urów

BjiHHHHe xuMimecKH HmyuapoBaHHoro ^H afieia Ha axTHBHOCTH COfl, nepoKCH^aau u Karajia3u y i<puc

Organs

SOD

U/g w . t . w . ; X +S U/g w . t . w . ; X +SP - a s e BU/g w .t.w , X +SC - a s e _ con-t r o l s d i a b e -t i c s con-t r o l s d ia b e -t i c s t r o l scon- d i a b e -t i c s 1 2 3 4 5 6 7 L i v e r 4 000 2 500 0 . 0 200 4 . 8 0 1 5 .9 0 +600 +300 +19 + 0 .4 3 + 1.2 1 Kidney 1 200 327 0 . 0 0 . 0 0 . 3 6 0 .9 2 6 +151 +41 + 0 .0 4 + 0 .1 0 0 S p le e n 560 272 963 2 810 2 . 4 0 0 .2 3 5 +50 +36 +91 +205 + 0.21 + 0 .0 4 0 T e s t e s 960 308 407 100 0 .4 1 0 .8 8 +63 +31 +41 + 9 .4 + 0 .0 3 + 0 .0 2 Whole b r a i n 240 185 120 160 0 .0 4 0 0 .0 3 8 +24 +22 +10 +12 + 0 .0 0 4 + 0 .0 1 0 Lung 210 241 872 1 070 0 .2 6 0 .2 1 0 +20 +26 +86 +100 + 0 .0 2 + 0 .0 2 0 P a n c re a s 310 128 136 280 0 .1 9 5 0 .1 1 7 +31 +10 +13 +30 + 0 .0 1 0 + 0 .0 0 7

T a b le 6 (c o n td .) 1 2 3 4 5 6 7 H ea rt 480 263 2 690 1 583 0 .2 4 5 0 .4 4 0 muscle +47 +29 +300 +96 + 0 .0 2 3 + 0 .0 5 0 S k e l e t a l 300 223 105 220 0 . 1 1 0 0 .1 6 0 m uscle _{+27 +30 +10 +24 + 0 .0 0 9 + 0 .0 1 0} H e m o ly sa t e* 696 525 11 666 1 172 2 .6 7 0 4 .1 4 0 +68 +50 +1 000 +131 + 0 .3 1 0 + 0 .3 9 0 * U / m l .

N

0

t e : SOD, P - a s e and C -a s e v a l u e s in normal r a t s ; e f f e c t on them o f c h e m i c a ll y -i n d u c e d d i a b e t e s (w .t.w . => wet t i s s u e w e i g h t ; n = 1 0 ) ( C - a s e a c t i v i t y v a l u e s a r e g iv e n a s b e f o r e ) .

T h i s i s i l l u s t r a t e d i n th e f o l l o w in g Tab. 6 , i n which t h e s e two a r t e r i e s , w ith t h e i r low SOD v a l u e s , d i f f e r from th e o t h e r two a r t e r i e s examined, th e pulmonary a r t e r y and th e t h o r a c i c aorta-.

T a b l e 7

B ovine f e t a l a r t e r i a l SOD SOD w t ę t n i c a c h płodowych c i e l ą t

00,3 b njionoBhix apTepHHX TejiHT

T i s s u e _{U/mg p r o t e i n}SOD U m b i li c a l a r t e r y 1 . 9 7 + 0 .1 6 Ductus a r t e r i o s u s 2 .3 2 + 0 . 3 3 T h o ra c ic a o r t a 3 .4 5 +0 .31 Pulmonary a r t e r y 3 .6 7 + 0 .3 2 At th e same ti m e , an i o n i z i n g r a d i a t i o n e f f e c t for mi ng th e oxygen r a d i c a l , f o r i n s t a n c e , c a u s e s a r t e r i o s c l e r o s i s i n expe-r im e n ta l a n im a l s [ 1 ] .

The enhanced tend enc y o f d i a b e t i c s t o a r t e r i o s c l e r o s i s can t h e r e f o r e be e x p l a i n e d on th e above b a s i s .

2 . E a r l y f o rm a ti o n o f c a t a r a c t i n d i a b e t i c s can s i m i l a r l y be w el l e x p l a i n e d i n th e l i g h t o f th e more r e c e n t e x a m i n a ti o n s . The l e n s o f th e eye i s p r o t e c t e d by SOD a g a i n s t 0 ^ , and by C -a se a g a i n s t ^-2 ^‘ A d e c r e a s e i n th e a c t i v i t y o f e i t h e r o f th e two enzymes would w e l l a c c o u n t f o r th e e a r l y c a t a r a c t i n d i a b e -t i c s . From t h i s a s p e c t c o m p a ra ti ve i n v e s t i g a t i o n s «h a v e been p erf orm ed on ly on t h e r a t r e t i n a [ 5 ]. I t would be i n t e r e s t i n g to compare t h e s e measurements f o r th e v a r i o u s p a r t s o f t h e r a t eye.

T a b l e 8

Comparison o f b lo o d and r e t i n a l SOD a c t i v i t y o f normal and d i a b e t i c r a t s

Porównanie ak ty w no śc i SOD w s ia t kó w c e i w krwi szcz urów normalnych i d ia b et y c z n y c h

CpaBHeHHe aKTHBHOCTH CO# B CemaTKe H B KpOBH HOpMaJIhHUX H 4HaÓ6THUX KpbIC T i s s u e SOD p q /g p r o t e i n n c o n t r o l d i a b e t i c R e ti n a 42 0 .8 7 0.6 1 + 0 .0 5 + 0 .0 3 B lood 12 8 1 . 2 5 1 .3 + 5 .1 +7 .1 C l e a r l y , t h e s e e x a m in a ti o n s s ho u ld b e , and w i l l be r e p e a te d on human e n u c l e a t e d eye and on e x t i r p a t e d c a t a r a c t i n d i a b e t i c s ( F i g . 3 ) . Much work i s s t i l l r e q u ir e d in t h e i n t e r e s t o f p r o v i d in g f u r t h e r e v id en c e i n f a v o u r o f th e a bove s u g g e s t i o n s . I f ou r h y p o t h e s i s mentioned i n th e i n t r o d u c t i o n i s t r u e in d i a b e t e s , th en th e a g i n g p r o c e s s o f th e c e l l and o t h e r d i f f e r e n -c e s o b se r v e d i n t h i s f i e l d -can a l s o be w el l e x p l a i n e d on th e b a s i s t h a t th e a c t i v i t i e s o f t h e o x i d a t i v e enzymes may d i f f e r from group t o g ro up . Thus, i n th o s e p o s s e s s i n g a h ig h e r o x i d a -t i v e enzyme a c -t i v i -t y , th e " f a s t b u r n e r s " , th e more i n t e n s i v e c e l l o x i d a t i v e damage l e a d s e a r l i e r t o th e a g i n g o f th e c e l l s and o f

th e e n t i r e o rg an i sm . However, l e t u s n o t d e a l w ith g e r o n to l o g y , b ut mention a few f a c t s :

1 . S y l v i a e t a l . [1 9 ] c o u l d d e m on s tra te a d i f f e r e n c e t o e l e c t r i c s t i m u l u s i n th e change o f r a t e o f m i c r o c i r c u l a t i o n and t e r m i n a l o x i d a t i o n ( t h e r e s p i r a t i o n c h a i n ) i n th e c e r e b r a l t i s s u e o f young and a g ed r a t s ( a sl o w e r r e g e n e r a t i v e a b i l i t y was d e t e c t e d i n th e ag ed r a t s ) . T h is i s n a t u r a l l y only an i s o l a t e d exam ple; we c o ul d a l s o r e f e r t o c e r t a i n o f o u r own i n v e s t i g a -t i o n s and o b s e r v a -t i o n s i n -t h i s c a s e -to o [ 1 2 ] . Of c o u r s e , t h e s e d i d n o t form s y s t e m a t i c ex a m in a ti on s r e l a t i n g t o o l d a g e .

2 . Many a u t h o r s r e f e r t o th e p u b l i c a t i o n by P a c k e r e t a l . [ 1 5 ] i n c o n n e c tio n w ith t h i s prob lem o f a g i n g . The t i t l e o f t h e i r work was " A n t i o x i d a n t s v e r s u s a g i n g " . . T h e i r f i n a l con-c l u s i o n was t h a t th e a n t i o x i d a n t s a r e s u b s t a n con-c e s o f s i m i l a r im p ortan c e t o th e v i t a m i n s : th ey p r o t e c t t h e org a n ism n o t only from a g i n g , b u t a l s o from tumorous damage, by i n h i b i t i n g th e f o rm a ti o n o f th e more s t r o n g l y c a r c i n o g e n i c i n t e r m e d i a t e s ; among o t h e r s , th ey p r o t e c t th e DNA from th e damage c a u s e d by p e r o x id e ( F i g . 4 ) . 2 0 2 Enlarging of 0 2 ♦ or Hj0 2 production 2 H (o r inhibition ot e nzy m e s ). H2 ° 2 ♦ H2 0 2 C~ as * » 0 2 ♦ 2 H 2 0 0 2 A H 2 -P-~ — » A ♦ H 2 0 F i g . 3 . C a t a r a c t o g e n e s i s Powstawanie k a t a r a k t y KaTapaKTor eHe3 L e t u s r e tu r n b r i e f l y t o th e f a c t t h a t in f la m m a tio n o c c u r s i n th e two s y st e m ic d i s e a s e s (r h eu m a tic d i s e a s e s and d i a b e t e s ) I mentioned p r e v i o u s l y . A c o n s i d e r a b l e p r o p o r ti o n o f th e i n f la m -matory p r o c e s s e s d e ve lo p on th e a c t i o n o f h i s ta m in e . I t emerged from th e o b s e r v a t i o n s o f 0 h m o r i e t a l . [ 1 3 ] t h a t th e x a n th i n e o x i d a s e - x a n t h i n e sy stem c a u s e s h i st a m i n e r e l e a s e from

C D _ON P re c a rc in o g e n C a rc in o g e n C a rc in o g e n + ta rg e t n u c le o p h il e o x id n . ( h y d r o x y la ti o n ) P o ly c y c li c s : 3 , 4 b e n z o p y re n e 9 , 1 0 - d im e . -1 , 2 d ib e n z a n -th r a c e n e 3 m e th y lc h o la n th re n e A ro m a ti c a m in e s : 2 a c e ty la m in o tl u o re n e 2 -n a p h ty la m in e A z o d y e s : p -d im e . a z o b e n z e n e U r e th a n e o x id n ./ re d u c tn . -* ■ ta rg e t c a rc in o g e n b io l. e x p re s s io n -+ ■ tu m o r in d u c ti o n A c ti v a ti o n c o u ld b e b io c h e m . e n z y m ic c h e m i -c a t e le c tr o c h e m . p h o to -c h e m . ra d ia ti o n re d u c ti o n N it ro th e te r o c y c le s a n ti b io ti c s : 4 n it r o q u in o li n e N - o x id e Q u in o n e s ', m it o m y c in C C IA F ig . 4 . R ed o x m o d e l fo r c h e m ic a l c a r c in o g e n e si s M o d e l re d o k s k a rc in o g e n e z y c h e m ic z n e j P ef lO K C -M O fl e^ b xH M H u ec K or o ic ap iy iH o re H e3 a Bela M a tk o vi cs

th e p e r i t o n e a l m ast c e l l s . The i n d u c t o r o f th e h is t a m in e r e l e a -s e i -s HgOg.

I n j e c t i o n o f + h is t a m in e g i v e s r i s e t o in fla m m a tory r a t paw oedema, which i s re du c ed by C - a s e . The PME t h e r e f o r e p la y im p o rt a n t r o l e s in th e development and a l s o th e d e c r e a s e o f i n -fl am m a tion . T h is i s p r ove d n ot only by th e r e l a t e d l i t e r a t u r e , b ut by numerous o t h e r d a t a t o o . F o r i n s t a n c e , dep end ing on t h e i r q u a n t i t i e s , t h e a n t i o h l o g i s t i c s a r e c a p a b l e o f d e c r e a s i n g th e Og a n ion p r o d u c t i o n [ 1 4 ] .

A lthough have m ention ed some a nim a l e x p e r im en ta l d a ta , p l e a s e a c c e p t my l e c t u r e a s p r i m a r i l y th o u g h t- p r ov o k in g w ith r e g a r d t o humans, a s I i n t en d e d i t . REFERENCES [ 1 ] A a r n o u d s e M. W. , L a m b e r t s H. B . , I n t . J . R a d i a t . B i o l . 3 1 , 87 ( 1 9 7 7 ). [ 2 ] B h u y a n K. C . , B h u y a D. K . , B iochim . B iop h y s. A cta 5 42 , 28 ( 1 9 7 8 ). [ 3 ] B r a y R. C. , C o c k l e S. A. , F l e l d e n E. M., R o b e r t s P. B . , R o t i l i o G. , C a l a b r e -s e L . , Biochem. J . 1 3 9 , 43 ( 1 9 7 4 ). [ 4 ] C a r r e l l R. W. , W i n t e r b o u r n C. C. , R a - c h m i l e w i t z E. A . , B r i t . J . H a e m a to l ., 3 0, 2 59 ( 1 9 7 5 ) . [ 5 ] C r o u c h R. , K i m s e y G. , P r i e s t D. G ., 3 a r d a A ., B u s e M. G ., D i a b e t o l o g i a , 15_, 53 (1 9 7 8 ) . [ 6 ] F r a z e r P. D. , B r a d y F . 0 . , P r oc . S o c . Exp. B i o l . Med. 1 59 , 30 (1 9 7 8 ) . [ 7 ] H a b e r F. , W e i s s J . , P ro c . Roy. S o c . L o n d ., S e r . A 1 47 , 332 (1 9 3 4 ) . [ 8 ] H u b e r W., S a i f e r M. G. P . , [ i n : ] S u p e r o -o x i d e and s u p e r o x i d e d is m u t a s e s (e d . M i c h e l s o n A. M. e t a l . ) London (1 9 7 7 ) . [ 9 ] M a t k o v i c s B . , Z e s z. Nauk. UL I I , 1 6, 103 (1 9 7 7 ) . [ 1 0 ] M a t k o v i c s B. , N o v a k R . , S z a b o ' L . , V a r g a S . I . , E x p e r i e n t . (u n d e r p u b l i c a t i o n ) ( 1 9 7 9 ) .

[1 1 ] M c C o r d J . M., S c i e n c e 1 0 5 , 529 (.1974). [1 2 ] N o v 4 k R . , M a t k o v i c s B. , M a r i k M., F a c h e t J . , E x p e r i e n t . 34 , 1134 (1 9 7 8 ) . [1 3 ] O h m o r i H. , K o m o r i y a K. , A z u m a A. , K u r o z u m l S . , H a s h i m o t o Y ., Biocherti. Phar-m ac ol . 2 8 , 333 ( 1 9 7 9 ).

[1 4 ] O y a n a g u l Y ., Biochem. Ph arm ac ol. 2 5 , 1473 (1 9 7 6 ). [1 5 ] P a c k e r L . , W a l t o n J . , CHEMTECH. 276 ( 1 9 7 7 ). [1 6 ] P u i g -P a r e l l a d a P. , P l a n a s J . M ., B

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[1 7 ] S c u d d e r P . , S t o c k s J . , D o r m a n d y T. L . , C l i n . Chim. A cta 6 9, 397 ( 1 9 7 6 ). [ 1 8 J S i n g h A ., Photochem. P h o t o b i o l . 2 8 , 429 (1 9 7 8 ) . [ 1 9 ] S y l v i a A. L . , R o s e n t h a l M., B r a i n R es. 1 6 5 , 235 (1 9 7 9 ). [ 2 0 ] W a l l i n g C ., A c c t s . Chem. R es. 8 , 125 ( 1 9 7 5 ). B i o l o g i c a l I s o t o p e L a b o r a t o r y Department o f B i o c h e m i st r y J o z s e f A t t i l a U n i v e r s i t y S z e g e d , Hungary B e l a M at kov ic s

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