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Protection Against Acetaldehyde Toxicity in The Rat by L-Cysteine, Thiamin and L-2-Methylthiazolidine-4-carboxylic Acid 1)
Protection Against Acetaldehyde Toxicity in The Rat by L-Cysteine, Thiamin and L-2-Methylthiazolidine-4-carboxylic Acid 1)
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Abstract
smoking (the two frequently occur in associationcould play a role in the development of cardiot
vascular disease [9]. From these observations, iis evident that studies of protection against acetal)
dehyde toxicity and lethality (preferably by naturally-occurring metabolites used in pharmacological doses) are very much in order.
Currently, on biochemical and biological
grounds there is a good basis for such studies.
On biochemical grounds, protection against
acetaldehyde might be obtained with two naturally-occurring compounds, L-cysteine and thiamin, either of which could complex with it by
way of free sulfhydry! (-SH) groups to eventually form relatively non-toxic compounds. Lcysteine could complex with acetaldehyde to
form a hemiacetal which would then cyclize to
form L-2-methylthiazolidine-4-carboxylic acid
(L-MTCA) [10, 11]. L-MTCA might then serve
as a non-toxic metabolic detoxification product
as well as a protective agent per se by generating
free intracellular (-SH) groups more effectively
as suggested by DEBEu et al. [11]. Thiamin as
cocarboxylase (pyrophosphate form) could complex with acetaldehyde by way of pyruvic oxidase
to give rise eventually by way of lipoic acid to
acetyl Coenzyme A [12]. On biological grounds,
both L-cysteine and L-MTCA could protect
against the lethal pulmonary edema induced by
acetaldehyde by way of free or latent (-SH)
groups in a manner similar to that reported
against the lethal pulmonary edema induced by
thiourea compounds [11, 13, 14].
1) Supported by US Veterans Administration (Project
Nos. 8078-01 and [8078] 2-70).
2) Address reprint requests to Dr. Herbert Sprince, VA
Hospital, Coatesville, Pennsylvania 19320, USA.
126
It is t h e p u r p o s e o f this p a p e r to p r e s e n t
d a t a o n t h e p r o t e c t i v e a c t i o n o f L-cysteine free
b a s e (FB), t h i a m i n . H C I , a n d L - M T C A a g a i n s t
a c e t a l d e h y d e t o x i c i t y a n d to c o m p a r e t h e p r o t e c t i v e d o s e levels w i t h t h e t o x i c d o s e levels o f
these compounds.
14, 16, 17, 18, 20 and 22. The calculated oral LD90 dose
level (Litchfield-Wilcoxon) was further checked experimentally in 50 rats 90 5 days old and weighing 365
20 g. The experimental oral LD90 thus determined was
the value taken as the standard lethal dose used hereafter
in this study. All calculated and experimental LD50 and
LD90 values are presented under Results. Reasons for
choice of the LD90 dose as the standard lethal dose are
presented under Discussion.
(d)
Results
(a)
A c e t a l d e h y d e : L D 50 and L D 90 values
T h e o r a l - 2 4 h o u r L D 50 a n d L D 9 0 v a l u e s
o f freshly distilled a c e t a l d e h y d e in m a l e C F E
rats, 90 =k 5 d a y s old, w e i g h i n g 365 i 20 g,
fasted o v e r n i g h t w e r e o b t a i n e d a n d c a l c u l a t e d b y
the L i t c h f i e l d - W i l c o x o n m e t h o d w i t h 19/20 c o n fidence limits [15] to be as f o l l o w s : L D 5 0 = 15.0
(14.4-15.6) m M / k g e q u i v a l e n t to 661 (634-687)
m g / k g a n d L D 9 0 = 17.8 (15.5-20.5) m M / k g
e q u i v a l e n t t o 784 ( 6 8 3 - 9 0 3 ) m g / k g . A d d i t i o n a l l y ,
w i t h an e x p e r i m e n t a l d o s e o f 18.0 m M / k g in
50 rats, t h e 24 h o u r l e t h a l r e s p o n s e was 45 rats
d e a d o u t o f 50 tested. H e n c e , 18.0 m M / k g
( = 793 m g / k g ) o f a c e t a l d e h y d e was u s e d herea f t e r as t h e s t a n d a r d l e t h a l d o s e ( o r a l 2 4 - h o u r
L D 90) in o u r p r o t e c t i o n e x p e r i m e n t s .
Toxic manifestations of acetaldehyde were
c h i e f l y r e s p i r a t o r y distress, g a s p i n g , a n d an
a n e s t h e t i c - l i k e p a r a l y s i s w i t h loss o f r i g h t i n g
reflexes as r e p o r t e d b y o t h e r s [3]. C h a r a c t e r i s t i cally, the a n e s t h e t i c ( h y p n o t i c ) effect o f acetald e h y d e d e v e l o p e d v e r y q u i c k l y after its i n t u b a t i o n ( w i t h i n 3 - 1 0 m i n u t e s ) a n d was o f s h o r t d u r a t i o n (5-15 m i n u t e s ) . T h e a n i m a l s a p p e a r e d to
r e c o v e r v e r y q u i c k l y a n d t o be n o r m a l f o r a b o u t
127
2-3 hours. Eventually, respiratory distress, stupor, and death ensued. Lethality was 54% in
3 hours and 90% in 24 hours (see Table 1), an
observation pointing to a delayed toxic effect.
Delayed lethality generally (but not always)
occurred in rats which had appeared to recover
from the anesthetic effect.
(b)
tion against the anesthetic effect generally resulted in protection against lethality. In separate
experiments (not shown in Table 1), protection
effects of L-cysteine FB and L - M T C A were not
appreciably increased at 3.0 m M / k g and were
decreased at 1.0 m M / k g . Thus, a dose o f 2.0 m M /
kg was the optimal protective dose o f these comp o u n d s under our test conditions.
Protection studies
Table 1
Protective action of L-cysteine FB (free base), thiamin 9HCI, and L-2-methylthiazolidine-4-carboxylic acid (L-MTCA)
against anesthetic and lethal effects of acetaldehyde (AcH) in the rat.
Compounds tested
AcH
3-10 minutes
3 hours
24 hours
72 hours
(oral dose)
(oral dose)
~ anesthetized
~ dead
~ dead
~ dead
18.0 mM/kg
18.0 mM/kg
96 (48/50)
20 (5/25)
54 (27/50)
8 (2/25)
90 (45/50)
20 (5/25)
90 (45/50)
20 (5/25)
18.0 mM/kg
25 (5/20)
5 (1/20)
25 (5/20)
25 (5/20)
18.0 mM/kg
10 (2/20)
0 (0/20)
10 (2/20)
10 (2/20)
18.0 mM/kg
73 (11/i5)
53 (8/15)
87 (13/15)
87 (13/15)
18.0 mM/kg
18.0 mM/kg
18.0 mM/kg
18.0 mM/kg
5 (1/20)
0 (0/15)
13 (2/15)
33 (5/t5)
0 (0/20)
0 (0/15)
0 (0/15)
27 (4/15)
0 (0/20)
13 (2/15)
13 (2/15)
33 (5/15)
0 (0/20)
20 (3/15)
13 (2/15)
33 (5/15)
Combinations of above
(F) (B + D)
(G) (C + D)
(H) (B+E)
(I) (C + E)
Compounds tested were intubated orally 3045 minutes prior to oral intubation of 18 mM/kg of AcH (LD90 dose).
Doses are expressed in millimoles/kilogram (mM/kg). After intubation, rats were observed over a 24-72 hour
period for anesthetic effects (loss of elevation and righting reflexes) and lethal response (deaths). Figures in parentheses = No. rats anesthetized or dead-No, rats tested. Anesthesia (loss of righting reflexes) with AcH generally
occurred within 3-10 minutes and lasted for 5-15 minutes. Deaths were recorded after 3, 24, and 72 hours.
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Discussion
Toxic dose levels of L-cysteine FB, L-MTCA and thiamin 9HC1 per se in the rat.
Compound tested
L-cysteine FB
LD50
LD 10
LD 1
L-MTCA
LD 50
LD 10
LD 1
Thiamin. HC1
LD50
LD 10
LD 1
mM/kg
mg/kg
TD/PD*)
15.6 (14.8-16.4)
13.8 (12.0-15.9)
11.5 (8.7-15.2)
1890 (1793-1987)
1672 (1454-1926)
1393 (1054-1842)
7.80
6.90
5.75
17.9 (17.0-18.8)
14.9 (13.1-17.0)
12.4 (9.7-15.8)
2635 (2502-2767)
2193 (1928-2502)
1825 (1428-2325)
8.95
7.45
6.20
11.0 (10.4-11.6)
8.6 (7.4-10.1)
6.6 ( 5.0- 8.8)
3710 (3508-3912)
2901 (2496-3407)
2226 (1686-2968)
5.5
4.3
3.3
LD values above are oral-24 hour Litchfield-Wilcoxon values obtained with male CFE rats, 90 4- 5 days old,
weighing 365 20 g and fasted overnight.
*) Ratio TD/PD = toxic dose-protective dose (2.0 mM/kg from Table 1). It measures the relative safety, i.e. the
number of times greater the toxic dose is than the optimally protective dose.
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References
[1] J. P. YON WARTBUR~, The Metabolism of Alcohol in
Normals and Alcoholics: Enzymes, in: The Biology of
Alcoholism, Vol. 1 (Eds B. Kissin and H. Begleiter;
Plenum Press, New York 1971), p. 63-102.
[2] J. R. NEWSOME, V. NORMAN and C. H. KEITH, Vapor
Phase Analysis of Cigarette Smoke, Tab. Sci. 9, 102110 (1965).
[3] J. AKABANE, Aldehydes and Related Compounds, in:
130