- Email: [email protected]
Putrescine has hypothermic and antipyretic activity, in rats
Life Sciences, Vol. 38, pp. 1293-1298 Printed in the U.S.A.
P U T R E S C I N E HAS
HYPOTHERMIC
Susanna Genedani,
AND
Pergamon Press
ANTIPYRETIC
ACTIVITY,
Mara Bernardi, Simonetta a n d A. B e r t o l i n i
IN RATS.
Tagliavini
I n s t i t u t e of P h a r m a c o l o g y , U n i v e r s i t y of M o d e n a , V i a G . C a m p i n. 287, I 4 1 1 0 0 M o d e n a , I t a l y (Received in final form January 29, 1986) Summary I n t r a p e r i t o n e a l i n j e c t i o n of p u t r e s c i n e i n d u c e d d o s e r e l a t e d h y p o t h e r m i a in r a t s . T h e e f f e c t was m o r e p r o n o u n c e d at r o o m t e m p e r a t u r e (22°C) t h a n in a w a r m e n v i r o n m e n t (30°C), the m a x i m u m h y p o t h e r m i a ( - 2 . 6 4 ± 0 . 2 9 ° C , 30 min. a f t e r t r e a t m e n t ) b e i n g o b t a i n e d w i t h the d o s e of 300 m g / K g a n d r e m a l n l n g s i g n i f i c a n t t h r o u g h o u t 3 hr of o b s e r v a t i o n . P u t r e s c i n e a l s o h a d a n t l p y r e t l c a c t i v i t y , as it s i g n i f i c a n t l y r e d u c e d p y r o g e n I n d u c e d f e v e r at a d o s e l e v e l (I00 m g / K g i.p.) i n e f f e c t i v e in c a u s i n g h y p o t h e r m l a in n o r m a l r a t s . The h y p o t h e r m i c a n d a n t i p y r e t i c e f f e c t s of p u t r e s c l n e w e r e not a s s o c i a t e d w l t h any o b v i o u s s i g n of t o x i c i t y . H y p o t h e r m i a is one of the e f f e c t s p r o d u c e d by p o l y a m i n e s ( p u t r e scine, s p e r m l d l n e a n d s p e r m i n e ) (1-4), a n d o c c u r s b o t h a f t e r intraperitoneal (i.p.) a n d a f t e r i n t r a c e r e b r o v e n t r i c u l a r (l.C.V.) i n j e c t i o n . It has r e c e n t l y b e e n r e p o r t e d t h a t t h e s e s u b s t a n c e s also have a potent anti-inflammatory e f f e c t b o t h in s e r o t o n i n - a n d in c a r r a g e n a a n - i n d u c e d o e d e m a (5-7). Taken together, these findings may logical interest. This prompted us effect of putrescine, and to study rimentally-induced hyperthermia, in not it has antipyretic activity as
render polyamlnes of pharmacoto characterize the hypothermic its possible influence on expeorder to ascertain whether or well.
Materlals and Methods A d u l t f e m a l e r a t s ( 2 0 0 - 2 4 0 g) of a W l s t a r s t r a i n ( M o r i n i , S . P o l o d'Enza, Reggio nell'Emllla, Italy), kept under standard conditions on a n a t u r a l l i g h t - d a r k c y c l e a n d fed ad l i b i t u m , w e r e u s e d ihroughout. Experiments (22 + 0.2°C)
were performed either in or xn a hot environment
a thermoneutral (30 + O.2°C).
0024-3205/86 9 3 . 0 0 + . 0 0 Copyright
(c)
1986 P e r g a m o n P r e s s
Ltd.
environment Body temperatu-
1294
Hypothermic
Effect of Putrescine
Vol. 38, No. 14, 1986
re w a s r e c o r d e d by a thermocouple the t i p of w h i c h w a s i n s e r t e d 45 m m i n t o the r e c t u m for 60 sec. T h e i n s t r u m e n t was repeatedly calibrated against a mercury thermometer in a c o n s t a n t temperature water bath. Fever was induced by the subcutaneous injection of a 12% s u s p e n sion of f e r m e n t e d y e a s t (I m l / l O 0 g / b . w . ) 9 hr b e f o r e the t e s t . Putrescine tonea]ly
(Merck-Schuchardt, Munchen, W.Germany) was intraperladministered at t h e d o s e s of 1 0 0 , 2 O O , 3 0 0 and 400 mg/Kg.
T h e b a s a l r e c t a l l e v e l w a s the m e a n of t h r e e v a l u e s , recorded before treatment or b e f o r e the y e a s t i n j e c t i o n . Thereafter, body temperature was recorded 30,60,120 a n d 180 m i n a f t e r t r e a t m e n t , between 8 a . m . and 1 p.m. Statistical analysis of d a t a multiple comparison test and sion.
was performed with ANOVA, followed by Dunnett's test, and by linear regres-
Results
As s h o w n in T a b l e I, the i n t r a p e r i t o n e a l injection of p u t r e s c i n e induced dose-related (up to 300 m g / K g ) h y p o t h e r m i a whlch depended in p a r t on a m b i e n t temperature. In a t h e r m o n e u t r a l environment (22°C) the maximum effectlve d o s e (300 m g / K g ) c a u s e d a f a l l of - 2 . 6 4 ± 0 . 2 9 ° C 30 m i n u t e s after treatment. At t h l s d o s e l e v e l the hypothermic effect remained statistlcally slgnificant throughout the 3 hr o b s e r v a t i o n period. Increaslng the d o s e f u r t h e r (400 mg/ Kg) n e i t h e r increased nor prolonged the h y p o t h e r m i c effect: on the contrary, the effect was weaker and shorter than that caused by the d o s e of 300 m g / K g . Putrescine also induced hypothermia dose-dependently in a w a r m e n v i r o n m e n t ( 3 0 ° C ) , b u t the e f f e c t w a s s l g n i f l c a n t l y weaker than t h a t i n d u c e d b y the s a m e d o s e s in a t h e r m o n e u t r a l environment ( 2 2 ° C ) , e x c e p t for the h l g h e s t dose (400 mg/Kg) which had similar e f f e c t s at b o t h a m b i e n t temperatures. As s h o w n in Fig. I, p u t r e s c i n e also reduced pyrogen-induced fever, and, in t h i s c a s e , the e f f e c t w a s s t r i c t l y dose-related, b o t h In I n t e n s i t y and d u r a t l o n (llnear r e g r e s s i o n ) . The mlnlmum effective d o s e (i00 m g / K g i . p . ) c a u s e d a r e d u c t i o n of f e v e r w h i c h r e m a l n e d significant f o r 30 to 120 m i n a f t e r i n j e c t i o n ; the antipyretic e f f e c t of 2 0 0 m g / K g of p u t r e s c i n e l a s t e d u n t i l the e n d of t h e observation period (180 m i n a f t e r t r e a t m e n t ) ; and body temperature was returned to %he b a s a l v a l u e s (or e v e n b e l o w ) b y the two hlghest d o s e s (500 a n d 400 m g / K g ) , and remained so t h r o u g h o u t the o b s e r v a tion period.
300 400
200
300 400
100 200
-1.22±0.31 -o.96±o.17
-1.02±0.29
-0.66±0.43
-1.43±0.45 -2.06±0.29 @@ -1.56i0.22*
-0.35±0.13 -o.18±o.38
60
-o.91±o.49 -O. 94_+0.16 •
-1.60±0.24 *@ -2.64±0.Z9* -1.58±0.25**
-0.30+0.07 -o.38_+o.31
30
ambient
temperatures.
-1.82±0.19
-0.54±0.31 • -1.62±0.38
-1.75±0.31* -2.02±0.20* -1.97±0. 18"
-0.20±0.07 -o.o3±o.45
12o
with
the same
-1.02±0.13
-o.58±o.34 •
-0.16±0.22
-1.70±0.27 ** -1.33±0.35
-0.65±0.43 -0.68±0.39
-O.2O±0.O7
~8o
at the f o l l o w i n g
at d l f f e r e n t ± S.E.
Changes in b o d y t e m p e r a t u r e (°C) times after t r e a t m e n t (min.):
i n j e c t i o n of p u t r e s c i n e 5 rats per group. Means
@ P < 0.01; * * P < 0.05 (ANOVA and D u n n e t t ' s test) c o m p a r e d w i t h controls. • P < 0.05 (ANOVA and m u l t i p l e c o m p a r l s o n test) c o m p a r e d with the v a l u e s o b t a i n e d treatment at the ambient t e m p e r a t u r e of 22°C.
Putrescine, Putrescine, Putrescine,
30
Treatment mg/Kg
22
(°C)
Ambment Temperature
of i n t r a p e r i t o n e a l At least
Saline Putrescine, Putrescine, Putrescine, Putrescine,
effect
Hypothermlc
TABLE 1
%0 %n
m 0
i--t
o
r-t
o
~D O
F-J ~D OO
LO OO
< O
1296
Hypothermic Effect of Putrescine
(j
Vol. 38, No. 14, 1986
+2
e
u i.
@
E
•
÷1
eO
A e--
0
@ c
m
L,-
0
i.a
n
|
60
treatment
I
i
120
180
rain FIG.
1
Effect of intraperltoneal Injection of saline; •= 100 mg/Kg; ~ = 200 mg/Kg; •= 400 mg/Kg) on pyrogen-lnduced fever. At per group. Means ± S.E. Ambient temperature (22 ± P< 0.01; e~ P< 0.05 (ANOVA, c o m p a r e d
putresclne ( • = 300 mg/Kg; [] = least 9 rats O.2°C) with controls).
Dlscusslon T h e p r e s e n t r e s u l t s s h o w that i n t r a p e r l t o n e a l i n j e c t i o n s of p u t r e scine induce hypothermia in r a t s in a d o s e - d e p e n d e n t f a s h i o n , the m i n i m u m e f f e c t l v e d o s e b e i n g 200 m g / K g . T h l s h y p o t h e r m l c effect d e p e n d s in part on the e n v i r o n m e n t a l temperature, for it is s[Kn]~icantly w e a k e r ( a l b e i t s t l l l p r e s e n t ) in a hot e n v l r o n m e n t (30°C) t h a n at r o o m t e m p e r a t u r e (22°C). This could mean that the mechanism of action of putrescine-lnduced hypothermia consists, at least in part.in ac~lvatlng tile heat-loss pathways.A direct effect on the central thermostat cannot,however, be completely :uled out, particularly in view of reports by other authors (3) attesting to a lowering of body temperature following the lntracerebroventrlcular injection of putresclne. Moreover, the present results show that putrescine has antlpyretlc activity, since it reduces pyrogen-induced fever even at a dose level (1OO
Vol. 38, No. 14, 1986
mg/kg) Whlch animals.
Hypothermic Effect of Putrescine
is i n e f f e c t i v e
in i n d u c i n g
hypothermia
1297
in n o r m a l
The p o s s i b l e m e c h a n i s m ( s ) of a c t i o n of p u t r e s c i n e in a f f e c t i n g thermoregulatlon can o n l y be a m a t t e r for s p e c u l a t i o n , for the d a t a p r e s e n t l y a v a i l a b l e s u g g e s t t h a t p u t r e s c i n e (and p o s s i b l y the o t h e r p o l y a m l n e s as w e l l ) m a y act in a v a r i e t y of ways: - it m a y act d i r e c t l y on the c e n t r a l t h e r m o - r e g u l a t o r y p a t h w a y s , e l t h e r as a s y n a p t i c t r a n s m i t t e r (8) or as a m o d u l a t o r (9); - o w i n g to xts p o l y c a t i o n i c n a t u r e , a n d in v i e w of r e c e n t in v i t r o s t u d i e s (I0) w h i c h s u g g e s t that m e m b r a n e p o t e n t i a l s r a t h e r t h a n s y n a p t i c e v e n t s m a y be r e s p o n s i b l e for the s e n s i t i v l t y of central n e u r o n s i n v o l v e d in t h e r m a l r e g u l a t l o n , p u t r e s c i n e m a y be thought to h a v e a d i r e c t e f f e c t on n e u r o n e m e m b r a n e p o t e n t i a l s ; s i n c e the h y p o t h e r m i c r o l e of h i s t a m i n e r g i c p a t h w a y s in the h y p o t h a l a m u s is w e l l k n o w n (11), R u t r e s c i n e - i n d u c e d hypothermia m a y be the r e s u l t of a r e l e a s e of h i s t a m i n e in the c e n t r a l n e r v o u s s y s t e m (CNS); s i n c e p o l y a m i n e s , i n t r o d u c e d i n t o m e d i a s u p p o r t i n g an i s o l a t e d organ system, can abolish transmembrane g l u c o s e t r a n s p o r t (12) a n d i n h i b i t a d e n y l c y c l a s e a n d N a ~ K ~ A T P a s e e n z y m e a c t i v i t y (1317), t h e l r h y p o t h e r m i c e f f e c t c o u l d be t h o u g h t to be due in p a r t to a s y s t e m i c i n h i b i t o r y e f f e c t on c e l l u l a r e n e r g e t i c s , as sugg e s t e d by C a m p b e l l et al. to e x p l a i n h y p o t h e r m i a in u r e m l c p a t l e n t s (4); - f i n a l l y , s i n c e p o l y a m i n e s i n h i b i t the a c t i v i t y of p h o s p h o l i p a s e A2 a n d p h o s p h o l i p a s e C (18), a n d s i n c e t h e y h a v e r e c e n t l y b e e n p r o v e d to be g l u c o c o r t i c o i d - t y p e anti-inflammatory d r u g s (7), t h e i r h y p o t h e r m i c e f f e c t m i g h t be due, at l e a s t in part, to i n h i b i t i o n of p r o s t a g l a n d i n s y n t h e s i s in the CNS. -
P o l y a m i n e s are d i s t r i b u t e d t h r o u g h o u t all l i v i n g t i s s u e , i n c l u d i n g n e r v o u s t i s s u e . T h e i r c o n t e n t is c l o s e l y r e g u l a t e d by the cell a n d is m a i n l y r e l a t e d to c e l l u l a r g r o w t h a n d d i f f e r e n t l a t l o n . P r e s u m a b l y , t h e s e c o m p o u n d s a l s o h a v e i m p o r t a n t f u n c t i o n s in n o n - g r o w i n g s y s t e m s , s u c h as the a d u l t m a m m a l i a n b r a i n , w h i c h is one of the r i c h e s t s o u r c e s of s p e r m i d i n e a n d s p e r m l n e s y n t h a s e s ( 1 9 ) . H o w e v e r , f r o m e x s i s t i n g p h a r m a c o l o g i c a l a n d b i o c h e m i c a l d a t a on b r a i n p o l y a m i n e s ( 2 0 , 2 1 ) , f u n c t i o n a l r o l e s s p e c i f i c for b r a i n c a n n o t be d e d u c e d . The p r e s e n t f l n d i n g s m i g h t i n d i c a t e t h a t p o l y a m i n e s p l a y a r o l e in t h e r m o - r e g u l a t i o n . Moreover, these same results,together w i t h r e c e n t r e p o r t s s h o w l n g that p o l y a m i n e s h a v e a n a l g e s i c (22) and anti-inflammatory (5-7) a c t i v i t i e s , m a y s u g g e s t for t h e s e c o m p o u n d s an u n e x p e c t e d r o l e as d r u g s . References 1. 6.6. 2. D.J.
Shaw, A r c h . i n t . P h a r m a c o d y n . 198, 3 6 - 4 8 (1972). A n d e r s o n , J. C r o s s l a n d a n d G.G. Shaw, N e u r o p h a r m a c o l . 571-577 (1975). 3. R.W. H o r t o n , J.F. C o l l i n s , G.M. A n l e z a r k a n d B.S. M e l d r u m ,
1_~,
1298
4.
5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19.
20. 21. 22.
Hypothermic Effect of Putrescine
Vol. 38, No. 14, 1986
Europ.J.Pharmacol. 5.__99, 7 5 - 8 3 ( 1 9 7 9 ) . R.Campbell, Y. T a l w a l k a r , D. B a r t o s , F. B a r t o s , J. Musgrave, M. H a r r i e r , H. P u r l , D. G r e t t i e , A.M. D o l n e y a n d B. L o g g a n , A d v a n c e s xn P o l y a m i n e R e s e a r c h e d s . R.A. C a m p b e l l , e t a l . , v o l . 2, pp. 3 1 9 - 3 4 3 , R a v e n P r e s s , New Y o r k ( 1 9 7 8 ) . J. Bird and B.A. Lewis, Br.J.Pharmacol. 7__4, 2 0 4 P - 2 0 5 P ( 1 9 8 1 ) . J . B i r d , S. M o h d - H i d z r a n d D . A . L e w i s , A g e n t s a n d A c t i o n s 1 3 , 342-347 (1983). Y. O y n a g u i , A g e n t s a n d A c t i o n s 1 4 , 2 2 8 - 2 3 7 ( 1 9 8 4 ) . 6 . 6 . Shaw, B i o c h e m . P h a r m a c o l . 26, 1450-1451 (1977). R . J . Harman a n d 6 . 6 . Shaw, B r . J . P h a r m a c o l . 73, 165-174 (1981). R.F. Hellon, Fed.Proc. 40, 2804-2806 (1981). P . Lomax a n d M.D. G r e e n , F e d . P r o c . 40, 2741-2745 (1981). S. A r v a n l t a k i s , J . M a n g o s , N . R . M c S h e r r y a n d O. R e n n e r t , Tex. Rep.Biol.Med. 3 4 , 175-186 (1976). K. Ahmed a n d H . G . W z l l l a m s - A s h m a n , B z o c h e m . J . 1 1 3 , 8 2 9 - 8 3 6 (1969). V . J . Atmar a n d G.D. Kuehn, F e d . P r o c . 3 6 , 686 ( 1 9 7 7 ) . 6 . Q u a r f o t h a n d K. Ahmed, F e d . P r o c . 3 6 , 360 ( 1 9 7 7 ) . Y. T a s h i m a a n d M. H a s e g a w a , B l o c h e m . B i o p h y s . R e s . C o m m . 66, 1344-1348 (1975). R. Wright, B.A. B u e h l e r and O.M. Rennert, P e d i a t r . R e s . 10, 373 (1976). A.M. Sechi, L. Cabrxnx, L. Landi, P. P a s q u a l l and 6. Lenaz, Arch.Biochem.Biophys. 186, 248-254 (1978). A. Raina, R.L. Pajula, T. E l o r a n t a and K. Tuomx, Advances in P o l y a m i n e R e s e a r c h eds. R.A. Campbell, D.R. Morrxs, D. Bartos, D. Daves and F. Bartos, vol. 1, pp. 75-82, Raven Press, New Y o r k (1978). G.6. Shaw, B i o c h e m . P h a r m a c o l . 28, 1-6 (1979). D.H. Russell, E. Feller, L.J. M a r t o n and S. Le Gendre, J. N e u r o b i o l . 5, 349-352 (1974). S. Genedanl, G. P i c c i n i n l and A. B e r t o l l n l , Llfe Scx. 24, 2 4 0 7 - 2 4 1 2 (1984).
P U T R E S C I N E HAS
HYPOTHERMIC
Susanna Genedani,
AND
Pergamon Press
ANTIPYRETIC
ACTIVITY,
Mara Bernardi, Simonetta a n d A. B e r t o l i n i
IN RATS.
Tagliavini
I n s t i t u t e of P h a r m a c o l o g y , U n i v e r s i t y of M o d e n a , V i a G . C a m p i n. 287, I 4 1 1 0 0 M o d e n a , I t a l y (Received in final form January 29, 1986) Summary I n t r a p e r i t o n e a l i n j e c t i o n of p u t r e s c i n e i n d u c e d d o s e r e l a t e d h y p o t h e r m i a in r a t s . T h e e f f e c t was m o r e p r o n o u n c e d at r o o m t e m p e r a t u r e (22°C) t h a n in a w a r m e n v i r o n m e n t (30°C), the m a x i m u m h y p o t h e r m i a ( - 2 . 6 4 ± 0 . 2 9 ° C , 30 min. a f t e r t r e a t m e n t ) b e i n g o b t a i n e d w i t h the d o s e of 300 m g / K g a n d r e m a l n l n g s i g n i f i c a n t t h r o u g h o u t 3 hr of o b s e r v a t i o n . P u t r e s c i n e a l s o h a d a n t l p y r e t l c a c t i v i t y , as it s i g n i f i c a n t l y r e d u c e d p y r o g e n I n d u c e d f e v e r at a d o s e l e v e l (I00 m g / K g i.p.) i n e f f e c t i v e in c a u s i n g h y p o t h e r m l a in n o r m a l r a t s . The h y p o t h e r m i c a n d a n t i p y r e t i c e f f e c t s of p u t r e s c l n e w e r e not a s s o c i a t e d w l t h any o b v i o u s s i g n of t o x i c i t y . H y p o t h e r m i a is one of the e f f e c t s p r o d u c e d by p o l y a m i n e s ( p u t r e scine, s p e r m l d l n e a n d s p e r m i n e ) (1-4), a n d o c c u r s b o t h a f t e r intraperitoneal (i.p.) a n d a f t e r i n t r a c e r e b r o v e n t r i c u l a r (l.C.V.) i n j e c t i o n . It has r e c e n t l y b e e n r e p o r t e d t h a t t h e s e s u b s t a n c e s also have a potent anti-inflammatory e f f e c t b o t h in s e r o t o n i n - a n d in c a r r a g e n a a n - i n d u c e d o e d e m a (5-7). Taken together, these findings may logical interest. This prompted us effect of putrescine, and to study rimentally-induced hyperthermia, in not it has antipyretic activity as
render polyamlnes of pharmacoto characterize the hypothermic its possible influence on expeorder to ascertain whether or well.
Materlals and Methods A d u l t f e m a l e r a t s ( 2 0 0 - 2 4 0 g) of a W l s t a r s t r a i n ( M o r i n i , S . P o l o d'Enza, Reggio nell'Emllla, Italy), kept under standard conditions on a n a t u r a l l i g h t - d a r k c y c l e a n d fed ad l i b i t u m , w e r e u s e d ihroughout. Experiments (22 + 0.2°C)
were performed either in or xn a hot environment
a thermoneutral (30 + O.2°C).
0024-3205/86 9 3 . 0 0 + . 0 0 Copyright
(c)
1986 P e r g a m o n P r e s s
Ltd.
environment Body temperatu-
1294
Hypothermic
Effect of Putrescine
Vol. 38, No. 14, 1986
re w a s r e c o r d e d by a thermocouple the t i p of w h i c h w a s i n s e r t e d 45 m m i n t o the r e c t u m for 60 sec. T h e i n s t r u m e n t was repeatedly calibrated against a mercury thermometer in a c o n s t a n t temperature water bath. Fever was induced by the subcutaneous injection of a 12% s u s p e n sion of f e r m e n t e d y e a s t (I m l / l O 0 g / b . w . ) 9 hr b e f o r e the t e s t . Putrescine tonea]ly
(Merck-Schuchardt, Munchen, W.Germany) was intraperladministered at t h e d o s e s of 1 0 0 , 2 O O , 3 0 0 and 400 mg/Kg.
T h e b a s a l r e c t a l l e v e l w a s the m e a n of t h r e e v a l u e s , recorded before treatment or b e f o r e the y e a s t i n j e c t i o n . Thereafter, body temperature was recorded 30,60,120 a n d 180 m i n a f t e r t r e a t m e n t , between 8 a . m . and 1 p.m. Statistical analysis of d a t a multiple comparison test and sion.
was performed with ANOVA, followed by Dunnett's test, and by linear regres-
Results
As s h o w n in T a b l e I, the i n t r a p e r i t o n e a l injection of p u t r e s c i n e induced dose-related (up to 300 m g / K g ) h y p o t h e r m i a whlch depended in p a r t on a m b i e n t temperature. In a t h e r m o n e u t r a l environment (22°C) the maximum effectlve d o s e (300 m g / K g ) c a u s e d a f a l l of - 2 . 6 4 ± 0 . 2 9 ° C 30 m i n u t e s after treatment. At t h l s d o s e l e v e l the hypothermic effect remained statistlcally slgnificant throughout the 3 hr o b s e r v a t i o n period. Increaslng the d o s e f u r t h e r (400 mg/ Kg) n e i t h e r increased nor prolonged the h y p o t h e r m i c effect: on the contrary, the effect was weaker and shorter than that caused by the d o s e of 300 m g / K g . Putrescine also induced hypothermia dose-dependently in a w a r m e n v i r o n m e n t ( 3 0 ° C ) , b u t the e f f e c t w a s s l g n i f l c a n t l y weaker than t h a t i n d u c e d b y the s a m e d o s e s in a t h e r m o n e u t r a l environment ( 2 2 ° C ) , e x c e p t for the h l g h e s t dose (400 mg/Kg) which had similar e f f e c t s at b o t h a m b i e n t temperatures. As s h o w n in Fig. I, p u t r e s c i n e also reduced pyrogen-induced fever, and, in t h i s c a s e , the e f f e c t w a s s t r i c t l y dose-related, b o t h In I n t e n s i t y and d u r a t l o n (llnear r e g r e s s i o n ) . The mlnlmum effective d o s e (i00 m g / K g i . p . ) c a u s e d a r e d u c t i o n of f e v e r w h i c h r e m a l n e d significant f o r 30 to 120 m i n a f t e r i n j e c t i o n ; the antipyretic e f f e c t of 2 0 0 m g / K g of p u t r e s c i n e l a s t e d u n t i l the e n d of t h e observation period (180 m i n a f t e r t r e a t m e n t ) ; and body temperature was returned to %he b a s a l v a l u e s (or e v e n b e l o w ) b y the two hlghest d o s e s (500 a n d 400 m g / K g ) , and remained so t h r o u g h o u t the o b s e r v a tion period.
300 400
200
300 400
100 200
-1.22±0.31 -o.96±o.17
-1.02±0.29
-0.66±0.43
-1.43±0.45 -2.06±0.29 @@ -1.56i0.22*
-0.35±0.13 -o.18±o.38
60
-o.91±o.49 -O. 94_+0.16 •
-1.60±0.24 *@ -2.64±0.Z9* -1.58±0.25**
-0.30+0.07 -o.38_+o.31
30
ambient
temperatures.
-1.82±0.19
-0.54±0.31 • -1.62±0.38
-1.75±0.31* -2.02±0.20* -1.97±0. 18"
-0.20±0.07 -o.o3±o.45
12o
with
the same
-1.02±0.13
-o.58±o.34 •
-0.16±0.22
-1.70±0.27 ** -1.33±0.35
-0.65±0.43 -0.68±0.39
-O.2O±0.O7
~8o
at the f o l l o w i n g
at d l f f e r e n t ± S.E.
Changes in b o d y t e m p e r a t u r e (°C) times after t r e a t m e n t (min.):
i n j e c t i o n of p u t r e s c i n e 5 rats per group. Means
@ P < 0.01; * * P < 0.05 (ANOVA and D u n n e t t ' s test) c o m p a r e d w i t h controls. • P < 0.05 (ANOVA and m u l t i p l e c o m p a r l s o n test) c o m p a r e d with the v a l u e s o b t a i n e d treatment at the ambient t e m p e r a t u r e of 22°C.
Putrescine, Putrescine, Putrescine,
30
Treatment mg/Kg
22
(°C)
Ambment Temperature
of i n t r a p e r i t o n e a l At least
Saline Putrescine, Putrescine, Putrescine, Putrescine,
effect
Hypothermlc
TABLE 1
%0 %n
m 0
i--t
o
r-t
o
~D O
F-J ~D OO
LO OO
< O
1296
Hypothermic Effect of Putrescine
(j
Vol. 38, No. 14, 1986
+2
e
u i.
@
E
•
÷1
eO
A e--
0
@ c
m
L,-
0
i.a
n
|
60
treatment
I
i
120
180
rain FIG.
1
Effect of intraperltoneal Injection of saline; •= 100 mg/Kg; ~ = 200 mg/Kg; •= 400 mg/Kg) on pyrogen-lnduced fever. At per group. Means ± S.E. Ambient temperature (22 ± P< 0.01; e~ P< 0.05 (ANOVA, c o m p a r e d
putresclne ( • = 300 mg/Kg; [] = least 9 rats O.2°C) with controls).
Dlscusslon T h e p r e s e n t r e s u l t s s h o w that i n t r a p e r l t o n e a l i n j e c t i o n s of p u t r e scine induce hypothermia in r a t s in a d o s e - d e p e n d e n t f a s h i o n , the m i n i m u m e f f e c t l v e d o s e b e i n g 200 m g / K g . T h l s h y p o t h e r m l c effect d e p e n d s in part on the e n v i r o n m e n t a l temperature, for it is s[Kn]~icantly w e a k e r ( a l b e i t s t l l l p r e s e n t ) in a hot e n v l r o n m e n t (30°C) t h a n at r o o m t e m p e r a t u r e (22°C). This could mean that the mechanism of action of putrescine-lnduced hypothermia consists, at least in part.in ac~lvatlng tile heat-loss pathways.A direct effect on the central thermostat cannot,however, be completely :uled out, particularly in view of reports by other authors (3) attesting to a lowering of body temperature following the lntracerebroventrlcular injection of putresclne. Moreover, the present results show that putrescine has antlpyretlc activity, since it reduces pyrogen-induced fever even at a dose level (1OO
Vol. 38, No. 14, 1986
mg/kg) Whlch animals.
Hypothermic Effect of Putrescine
is i n e f f e c t i v e
in i n d u c i n g
hypothermia
1297
in n o r m a l
The p o s s i b l e m e c h a n i s m ( s ) of a c t i o n of p u t r e s c i n e in a f f e c t i n g thermoregulatlon can o n l y be a m a t t e r for s p e c u l a t i o n , for the d a t a p r e s e n t l y a v a i l a b l e s u g g e s t t h a t p u t r e s c i n e (and p o s s i b l y the o t h e r p o l y a m l n e s as w e l l ) m a y act in a v a r i e t y of ways: - it m a y act d i r e c t l y on the c e n t r a l t h e r m o - r e g u l a t o r y p a t h w a y s , e l t h e r as a s y n a p t i c t r a n s m i t t e r (8) or as a m o d u l a t o r (9); - o w i n g to xts p o l y c a t i o n i c n a t u r e , a n d in v i e w of r e c e n t in v i t r o s t u d i e s (I0) w h i c h s u g g e s t that m e m b r a n e p o t e n t i a l s r a t h e r t h a n s y n a p t i c e v e n t s m a y be r e s p o n s i b l e for the s e n s i t i v l t y of central n e u r o n s i n v o l v e d in t h e r m a l r e g u l a t l o n , p u t r e s c i n e m a y be thought to h a v e a d i r e c t e f f e c t on n e u r o n e m e m b r a n e p o t e n t i a l s ; s i n c e the h y p o t h e r m i c r o l e of h i s t a m i n e r g i c p a t h w a y s in the h y p o t h a l a m u s is w e l l k n o w n (11), R u t r e s c i n e - i n d u c e d hypothermia m a y be the r e s u l t of a r e l e a s e of h i s t a m i n e in the c e n t r a l n e r v o u s s y s t e m (CNS); s i n c e p o l y a m i n e s , i n t r o d u c e d i n t o m e d i a s u p p o r t i n g an i s o l a t e d organ system, can abolish transmembrane g l u c o s e t r a n s p o r t (12) a n d i n h i b i t a d e n y l c y c l a s e a n d N a ~ K ~ A T P a s e e n z y m e a c t i v i t y (1317), t h e l r h y p o t h e r m i c e f f e c t c o u l d be t h o u g h t to be due in p a r t to a s y s t e m i c i n h i b i t o r y e f f e c t on c e l l u l a r e n e r g e t i c s , as sugg e s t e d by C a m p b e l l et al. to e x p l a i n h y p o t h e r m i a in u r e m l c p a t l e n t s (4); - f i n a l l y , s i n c e p o l y a m i n e s i n h i b i t the a c t i v i t y of p h o s p h o l i p a s e A2 a n d p h o s p h o l i p a s e C (18), a n d s i n c e t h e y h a v e r e c e n t l y b e e n p r o v e d to be g l u c o c o r t i c o i d - t y p e anti-inflammatory d r u g s (7), t h e i r h y p o t h e r m i c e f f e c t m i g h t be due, at l e a s t in part, to i n h i b i t i o n of p r o s t a g l a n d i n s y n t h e s i s in the CNS. -
P o l y a m i n e s are d i s t r i b u t e d t h r o u g h o u t all l i v i n g t i s s u e , i n c l u d i n g n e r v o u s t i s s u e . T h e i r c o n t e n t is c l o s e l y r e g u l a t e d by the cell a n d is m a i n l y r e l a t e d to c e l l u l a r g r o w t h a n d d i f f e r e n t l a t l o n . P r e s u m a b l y , t h e s e c o m p o u n d s a l s o h a v e i m p o r t a n t f u n c t i o n s in n o n - g r o w i n g s y s t e m s , s u c h as the a d u l t m a m m a l i a n b r a i n , w h i c h is one of the r i c h e s t s o u r c e s of s p e r m i d i n e a n d s p e r m l n e s y n t h a s e s ( 1 9 ) . H o w e v e r , f r o m e x s i s t i n g p h a r m a c o l o g i c a l a n d b i o c h e m i c a l d a t a on b r a i n p o l y a m i n e s ( 2 0 , 2 1 ) , f u n c t i o n a l r o l e s s p e c i f i c for b r a i n c a n n o t be d e d u c e d . The p r e s e n t f l n d i n g s m i g h t i n d i c a t e t h a t p o l y a m i n e s p l a y a r o l e in t h e r m o - r e g u l a t i o n . Moreover, these same results,together w i t h r e c e n t r e p o r t s s h o w l n g that p o l y a m i n e s h a v e a n a l g e s i c (22) and anti-inflammatory (5-7) a c t i v i t i e s , m a y s u g g e s t for t h e s e c o m p o u n d s an u n e x p e c t e d r o l e as d r u g s . References 1. 6.6. 2. D.J.
Shaw, A r c h . i n t . P h a r m a c o d y n . 198, 3 6 - 4 8 (1972). A n d e r s o n , J. C r o s s l a n d a n d G.G. Shaw, N e u r o p h a r m a c o l . 571-577 (1975). 3. R.W. H o r t o n , J.F. C o l l i n s , G.M. A n l e z a r k a n d B.S. M e l d r u m ,
1_~,
1298
4.
5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19.
20. 21. 22.
Hypothermic Effect of Putrescine
Vol. 38, No. 14, 1986
Europ.J.Pharmacol. 5.__99, 7 5 - 8 3 ( 1 9 7 9 ) . R.Campbell, Y. T a l w a l k a r , D. B a r t o s , F. B a r t o s , J. Musgrave, M. H a r r i e r , H. P u r l , D. G r e t t i e , A.M. D o l n e y a n d B. L o g g a n , A d v a n c e s xn P o l y a m i n e R e s e a r c h e d s . R.A. C a m p b e l l , e t a l . , v o l . 2, pp. 3 1 9 - 3 4 3 , R a v e n P r e s s , New Y o r k ( 1 9 7 8 ) . J. Bird and B.A. Lewis, Br.J.Pharmacol. 7__4, 2 0 4 P - 2 0 5 P ( 1 9 8 1 ) . J . B i r d , S. M o h d - H i d z r a n d D . A . L e w i s , A g e n t s a n d A c t i o n s 1 3 , 342-347 (1983). Y. O y n a g u i , A g e n t s a n d A c t i o n s 1 4 , 2 2 8 - 2 3 7 ( 1 9 8 4 ) . 6 . 6 . Shaw, B i o c h e m . P h a r m a c o l . 26, 1450-1451 (1977). R . J . Harman a n d 6 . 6 . Shaw, B r . J . P h a r m a c o l . 73, 165-174 (1981). R.F. Hellon, Fed.Proc. 40, 2804-2806 (1981). P . Lomax a n d M.D. G r e e n , F e d . P r o c . 40, 2741-2745 (1981). S. A r v a n l t a k i s , J . M a n g o s , N . R . M c S h e r r y a n d O. R e n n e r t , Tex. Rep.Biol.Med. 3 4 , 175-186 (1976). K. Ahmed a n d H . G . W z l l l a m s - A s h m a n , B z o c h e m . J . 1 1 3 , 8 2 9 - 8 3 6 (1969). V . J . Atmar a n d G.D. Kuehn, F e d . P r o c . 3 6 , 686 ( 1 9 7 7 ) . 6 . Q u a r f o t h a n d K. Ahmed, F e d . P r o c . 3 6 , 360 ( 1 9 7 7 ) . Y. T a s h i m a a n d M. H a s e g a w a , B l o c h e m . B i o p h y s . R e s . C o m m . 66, 1344-1348 (1975). R. Wright, B.A. B u e h l e r and O.M. Rennert, P e d i a t r . R e s . 10, 373 (1976). A.M. Sechi, L. Cabrxnx, L. Landi, P. P a s q u a l l and 6. Lenaz, Arch.Biochem.Biophys. 186, 248-254 (1978). A. Raina, R.L. Pajula, T. E l o r a n t a and K. Tuomx, Advances in P o l y a m i n e R e s e a r c h eds. R.A. Campbell, D.R. Morrxs, D. Bartos, D. Daves and F. Bartos, vol. 1, pp. 75-82, Raven Press, New Y o r k (1978). G.6. Shaw, B i o c h e m . P h a r m a c o l . 28, 1-6 (1979). D.H. Russell, E. Feller, L.J. M a r t o n and S. Le Gendre, J. N e u r o b i o l . 5, 349-352 (1974). S. Genedanl, G. P i c c i n i n l and A. B e r t o l l n l , Llfe Scx. 24, 2 4 0 7 - 2 4 1 2 (1984).