Z. Ern&hrungswiss.18, 184-190 (1979) 9 1979 Dr. Dietrich Steinkopff Verlag, Darmstadt ISSN 0044-264 X
Institute of Nutrition, Budapest (Hungary) (Director: Prof. Dr. R. Tarj~n)
H e p a t i c m e t a b o l i s m in e t h a n o l - t r e a t e d y o u n g rats M. Antal,
O. G a ~ l , A . G e r g e l y ,
K. Nagy,
and J. Sz~pvdlgyi
With 5 tables ( R e c e i v e d F e b r u a r y 26, 1979)
T h e n u m b e r of p r e g n a n t w o m e n , l a c t a t i n g m o t h e r s a n d y o u n g p e o p l e i n g e s t i n g e t h a n o l r e g u l a r l y is i n c r e a s i n g y e a r b y y e a r . S c a r c e d a t a a r e o n l y available on the effects of ethanol on physiologic functions in the developi n g o r g a n i s m e i t h e r i n a n i m a l s o r i n h u m a n b e i n g s (7, 14, 27). In the present series on investigations we studied the effect of ethanol on some metabolic processes of the developing organism in rats ingesting ethanol from delivery (via the breast milk) or from weanling.
Materials and methods D a m s of a W i s t a r - d e r i v e d strain, b r e d in o u r i n s t i t u t e s i n c e 1949 (OETI strain) w e r e o f f e r e d a 15% a q u e o u s e t h a n o l s o l u t i o n as d r i n k i n g fluid i m m e d i a t e l y a f t e r d e l i v e r y u n t i l w e a n l i n g . W e a n l i n g w a s t h e 28th d a y a f t e r delivery. T h e i r o f f s p r i n g s also r e c e i v e d t h e s a m e d r i n k i n g - f l u i d f r o m w e a n l i n g ( G r o u p A). A n o t h e r g r o u p of r a t s r e c e i v e d t h e 15% a q u e o u s e t h a n o l s o l u t i o n o n l y a f t e r w e a n l i n g ( G r o u p B). S i b l i n g s of r a t s i n g r o u p B r e c e i v e d t a p w a t e r a n d s e r v e d as c o n t r o l s ( G r o u p C). B o t h t h e 15% a q u e o u s e t h a n o l a n d t a p w a t e r w e r e o f f e r e d a d l i b i t u m . A n i m a l s w e r e f e d a n o r m a l r a t ' s c h o w (LATI, 369 k c a l p e r 100 g). T h e i n d i v i d u a l f l u i d a n d f o o d c o n s u m p t i o n a n d b o d y w e i g h t s w e r e m e a s u r e d at i n t e r v a l s . A few d a y s p r i o r to sacrifice h e m a t o c r i t w a s d e t e r m i n e d i n t h e tail b l o o d u s i n g an AB Lars-Ljunberg hematocrit centrifuge. All of t h e a n i m a l s w e r e s a c r i f i c e d b y d e c a p i t a t i o n a t t h e e n d of t h e 12th w e e k . SGOT and SGPT activities were assayed by Galenopharm testkit. Livers were p r o m p t l y r e m o v e d a n d w e i g h e d . P a r a m e t e r s for h e p a t i c m e t a b o l i s m w e r e s t u d i e d in t h e l i v e r s of 10-10 rats. H e p a t i c g l y c o g e n c o n t e n t w a s e s t i m a t e d b y t h e m e t h o d of Good et al. (12) u s i n g o - t o l u i d i n e r e a g e n t (4). T o t a l p r o t e i n c o n t e n t w a s d e t e r m i n e d b y K j e l d a h l ' s m e t h o d . H e p a t i c t r i g l y c e r i d e s w e r e e x t r a c t e d b y t h e t e c h n i q u e of Folcb et al. (8) a n d q u a n t i t a t e d b y B o e h r i n g e r U V test. L i v e r s a m p l e s (1 g/10 ml) w e r e h o m o g e n i z e d in a P o t t e r - E l v e h j e m t y p e t e f l o n g l a s s - t u b e in 0.25 M s u c r o s e m e d i u m . T h e h o m o g e n a t e w a s c e n t r i f u g e d at 10,000 • g for 15 r a i n i n a B e c k m a n c e n t r i f u g e (J-21B). T h e s u p e r n a t a n t w a s s p u n at 105,000 • g for 60 m i n in a n u l t r a c e n t r i f u g e ( J a n e t z k i V a c 601). G l u c o s e - 6 - p h o s p h a t e d e h y d r o g e n a s e ( G 6 P - D H ) w a s a s s a y e d i n t h e 105,000 • g s u p e r n a t a n t b y t h e m e t h o d of Glock a n d McLean (10) as m o d i f i e d b y Ldhrand Waller(20), a l c o h o l d e h y d r o g e n a s e (ADH) w a s a s s a y e d a c c o r d i n g to t h e m e t h o d d e s c r i b e d b y BDeher a n d RedetzRi (6). I n t h e G 6 P - D H or A D H a c t i v i t y a s s a y s t h e k i n e t i c s of t h e g e n e r a t i o n of N A D P H or N A D H w a s f o l l o w e d in a U n i c a m A C 30 A u t o m a t i c c h e m i s t r y s y s t e m at 340 n m a n d 25 ~ G l u c o s e - 6 - p h o s p h a t a s e (G6P-ase) a c t i v i t y w a s d e t e r m i n e d i n t h e 105,000 • g s e d i m e n t b y t h e m e t h o d of Baginski (3). 565
37 a'b' 73 71
39 a'b' 72 71
Male A B C
Female A B C
70 a'b' 95 ~' 128
81 a'b' 114 ~' 143
2
90 a'b' 112 ~' 156
101 a'b' 138 c' 166
3
114 a'b' 142 r 180
131 a'b' 175 188
4
152a'b ' 177 c' 203
168a'b ' 191 ~' 222
224a'b ' 248 ~ 291 181a'b ' 194 r 226
239a'b ' 262 ~' 300
8
192a'b ' 210 ~' 234
254a'b ' 285 r 306
9
195a'b' 218 r 241
265 a' 292 ~ 315
10
Significance
p
Institute of Nutrition, Budapest (Hungary) (Director: Prof. Dr. R. Tarj~n)
H e p a t i c m e t a b o l i s m in e t h a n o l - t r e a t e d y o u n g rats M. Antal,
O. G a ~ l , A . G e r g e l y ,
K. Nagy,
and J. Sz~pvdlgyi
With 5 tables ( R e c e i v e d F e b r u a r y 26, 1979)
T h e n u m b e r of p r e g n a n t w o m e n , l a c t a t i n g m o t h e r s a n d y o u n g p e o p l e i n g e s t i n g e t h a n o l r e g u l a r l y is i n c r e a s i n g y e a r b y y e a r . S c a r c e d a t a a r e o n l y available on the effects of ethanol on physiologic functions in the developi n g o r g a n i s m e i t h e r i n a n i m a l s o r i n h u m a n b e i n g s (7, 14, 27). In the present series on investigations we studied the effect of ethanol on some metabolic processes of the developing organism in rats ingesting ethanol from delivery (via the breast milk) or from weanling.
Materials and methods D a m s of a W i s t a r - d e r i v e d strain, b r e d in o u r i n s t i t u t e s i n c e 1949 (OETI strain) w e r e o f f e r e d a 15% a q u e o u s e t h a n o l s o l u t i o n as d r i n k i n g fluid i m m e d i a t e l y a f t e r d e l i v e r y u n t i l w e a n l i n g . W e a n l i n g w a s t h e 28th d a y a f t e r delivery. T h e i r o f f s p r i n g s also r e c e i v e d t h e s a m e d r i n k i n g - f l u i d f r o m w e a n l i n g ( G r o u p A). A n o t h e r g r o u p of r a t s r e c e i v e d t h e 15% a q u e o u s e t h a n o l s o l u t i o n o n l y a f t e r w e a n l i n g ( G r o u p B). S i b l i n g s of r a t s i n g r o u p B r e c e i v e d t a p w a t e r a n d s e r v e d as c o n t r o l s ( G r o u p C). B o t h t h e 15% a q u e o u s e t h a n o l a n d t a p w a t e r w e r e o f f e r e d a d l i b i t u m . A n i m a l s w e r e f e d a n o r m a l r a t ' s c h o w (LATI, 369 k c a l p e r 100 g). T h e i n d i v i d u a l f l u i d a n d f o o d c o n s u m p t i o n a n d b o d y w e i g h t s w e r e m e a s u r e d at i n t e r v a l s . A few d a y s p r i o r to sacrifice h e m a t o c r i t w a s d e t e r m i n e d i n t h e tail b l o o d u s i n g an AB Lars-Ljunberg hematocrit centrifuge. All of t h e a n i m a l s w e r e s a c r i f i c e d b y d e c a p i t a t i o n a t t h e e n d of t h e 12th w e e k . SGOT and SGPT activities were assayed by Galenopharm testkit. Livers were p r o m p t l y r e m o v e d a n d w e i g h e d . P a r a m e t e r s for h e p a t i c m e t a b o l i s m w e r e s t u d i e d in t h e l i v e r s of 10-10 rats. H e p a t i c g l y c o g e n c o n t e n t w a s e s t i m a t e d b y t h e m e t h o d of Good et al. (12) u s i n g o - t o l u i d i n e r e a g e n t (4). T o t a l p r o t e i n c o n t e n t w a s d e t e r m i n e d b y K j e l d a h l ' s m e t h o d . H e p a t i c t r i g l y c e r i d e s w e r e e x t r a c t e d b y t h e t e c h n i q u e of Folcb et al. (8) a n d q u a n t i t a t e d b y B o e h r i n g e r U V test. L i v e r s a m p l e s (1 g/10 ml) w e r e h o m o g e n i z e d in a P o t t e r - E l v e h j e m t y p e t e f l o n g l a s s - t u b e in 0.25 M s u c r o s e m e d i u m . T h e h o m o g e n a t e w a s c e n t r i f u g e d at 10,000 • g for 15 r a i n i n a B e c k m a n c e n t r i f u g e (J-21B). T h e s u p e r n a t a n t w a s s p u n at 105,000 • g for 60 m i n in a n u l t r a c e n t r i f u g e ( J a n e t z k i V a c 601). G l u c o s e - 6 - p h o s p h a t e d e h y d r o g e n a s e ( G 6 P - D H ) w a s a s s a y e d i n t h e 105,000 • g s u p e r n a t a n t b y t h e m e t h o d of Glock a n d McLean (10) as m o d i f i e d b y Ldhrand Waller(20), a l c o h o l d e h y d r o g e n a s e (ADH) w a s a s s a y e d a c c o r d i n g to t h e m e t h o d d e s c r i b e d b y BDeher a n d RedetzRi (6). I n t h e G 6 P - D H or A D H a c t i v i t y a s s a y s t h e k i n e t i c s of t h e g e n e r a t i o n of N A D P H or N A D H w a s f o l l o w e d in a U n i c a m A C 30 A u t o m a t i c c h e m i s t r y s y s t e m at 340 n m a n d 25 ~ G l u c o s e - 6 - p h o s p h a t a s e (G6P-ase) a c t i v i t y w a s d e t e r m i n e d i n t h e 105,000 • g s e d i m e n t b y t h e m e t h o d of Baginski (3). 565
37 a'b' 73 71
39 a'b' 72 71
Male A B C
Female A B C
70 a'b' 95 ~' 128
81 a'b' 114 ~' 143
2
90 a'b' 112 ~' 156
101 a'b' 138 c' 166
3
114 a'b' 142 r 180
131 a'b' 175 188
4
152a'b ' 177 c' 203
168a'b ' 191 ~' 222
224a'b ' 248 ~ 291 181a'b ' 194 r 226
239a'b ' 262 ~' 300
8
192a'b ' 210 ~' 234
254a'b ' 285 r 306
9
195a'b' 218 r 241
265 a' 292 ~ 315
10
Significance
p
