Interactions between ethanol and amphetamine in mice and rats

prog.

Neuro-Psychophamnco

Pergamon

Press

a, i 9 80.

1. v0 1.4 ,pp . I 3-I

Ltd.Printed

in

INTEUCI’IONS

Great

Britain.

BETWEEN ETHANOL AND MICE ANTI RATS ODILE

SPREUK-VAROQUAUX

Facult6

DGpartement de MCdecine Paris,

(Final

and

de

form,

PIERRE

AMI’HlRAlWNE

IN

SIMON

Pharmacologic Piti&SalpgtriPre France

December

1979)

Abstract

1.

Interactions macological Toxicity of Hyperthermia by ethanol. Stereotyped administration. These effects

2. 3. 4. 5. Keywords: stereotyped

between ethanol tests. d-amphetamine in and hypermotility behaviour were

ethanol, behaviour.

and

aggregated produced

produced

by

observed

in

d-amphetamine,

amphetamine

were

toxicity

on

mice was reduced by d-amphetamine

d-amphetamine

about

studied

-1

2 g.kg in

in

rats

acute

aggregated

or in

four

mice,

psychophar-

eliminated mice were

by ethanol. partly suppressed

unchanged

after

was

dose

different

of

ethanol

given

hyperthermia,

ethanol orally.

hypermotility,

Introduction In animal and man, several divergent various tests measuring motor, cognitive ions and conditioned task were reported. In animal effects of

, effects amphetamine

- stereotyped - hypermotility - improvement 1976). Toxicity 1974) were

Some

of amphetamine were increased:

behaviour (Todq of the

in isolated not modified.

Similarly

effects effects

of

- hypothermia - sleeping - impairment

time of

Other effects - open-field - discriminative

were

modified

in

(Todzy and Becker, 1974), and Becker, 1974)) acquisition of a conditioned

mice (Iverson Toxicity in of

in

interactions between and discriminative

ethanol

were

et al., rabbits modified

response

were

gerbils in rats rota-rod

(Jzrbe and Ohlin, 1977), (Breese et al., 1974), performance (Rech et al.,

in

hypermotility 1941)

different

ethanol

of ethanol measurements effects

different

1975) and (Reifenstein, in

ethanol functions,

directions

increased:

were not modified in gerbils (X&be and in a T-maze in gerbils

13

1976).

Ohlin, (J&be,

1977), 1977).

and amphetamine behavioural

on modificat-

directions

by

ethanol

. Some

a shuttle

box

(Merle

et

al.,

(Breese

et

al.,

was

in rats reduced. by

ethanol.

0. Spreux-Varoquaux

14 Finally, : -

some effects

of ethanol

and P. Simon

were reduced:

hypothermia in mice (Abdallah and Roby, 1975). (Reifenetein, sleeping time in mice (Abdallah and Roby, 1975) and in rabbits decrease of exploration in a T-mace in rats (Leonard and Wiseman, 1970), discriminative ability in a shuttle-box in rats (Schechter, 1974).

1941),

and skill tasks produced by ethanol were not In man various impairments of motor, attention modified by amphetamine (Kaplan et al., 1966; Brown et al., 1966; Forney et al., 1964; Hughes 1966) but some impairments of learning and Forney, 1964; Taylor et al., 1964; Wilson et al., and performance of mental calculation were suppressed by amphetamine(Taylor et a1.,1964;Wilson et al., 1966). The purpose of the present study was to assess interactions of ethanol with amphetamine on some psychopharmacological tests in mice and rats. Methods a. Material

and experimental

conditions

Wale Swiss WMRI mice weighing 20 to 25 g and male Wistar (AF) rats weighing 180 to 220 g were obtained from a homogeneous breeding. The animals were fasted 18 hours before the experiments. Ethanol was administered in one single oral dose and amphetamine in i.p. injection (0.25 ml per 100 g body weight in rats). of solution per 20 g body weight in mice, 0.50 ml of solution The hydroalcoholic solutions were always prepared just before use. Control animals always received distilled water in the same experimental conditions as the substances studied. Experiments were always performed at room temperature ranged from 20 to 22OC between 9 a.m. and 5 p.m. and conducted in blind conditions. b. Behavioural Stereotypies

tests

by d-amphetamine in rats.Rats (6 per plyup) received an i.p. injection of d-amphetamine sulfate (2 mg.kg ) 30 minutes after a given orally dose of ethanol. Then they were immediately individually placed in a plastic box (20 x 10 x 10 cm). Stereotyped behaviour of the head and of the buccal area were blindly scored from 0 to 3 according to their intensity at 10 minutes intervals until disappearance. Toxicity - First amphetamine placed in intervals died were

induced

of d-amphetamine in aggregated mice. experiment: a givexrldose of ethanol was administered orally 30 minutes before dsulfate (8 mg.kg i.p.). Following the injection of d-amphetamine the mice were groups of 10 in a plastic cage (20 x 10 x 10 cm). Deaths were recorded at hourly during 5 hours and 24 hours after the d-amphetamine administration. Animals which removed from the box at the same times.

- Second experiment: kinetic study of this effect. Ethanol (3 g.kg-')_yas orally 30, 60, 120 or 240 minutes before d-amphetamine sulfate (8 mg.kg Deaths were recorded in the same conditions as above.

administered i.p.) injection.

Hyperthermia produced by d-amphetamine in mice.- The experimental conditions were as for the toxicity of d-amphetamine in aggregated mice test. D-amphetamine sulfate i.p.) was administered 30 minutes after an oral administration of ethanol. Following injection of d-amphetamine, the animals were placed in groups of 10 in plastic cages (20 x 10 x 10 cm). Rectal temperature was recorded at 30 minutes intervals after the of d-amphetamine during 120 minutes. Mortality was observed at the same times.

the same-, (8 mg.kg the injection

Wypenwtility produced by d-amphetamine in mice.- D-amphetamine sulfate (3.5 mg.kg-'i.e.) was injected 15 mrnutes before a given dose of ethanol orally administered. The animals were placed 30 minutes after the administration of d-amphetamine in a photoelectric cells actimeter described by Boissier andSimon (1965). The number of crossed beams was recorded at 10 minutes intervals during 60 minutes. c. Statistical

analysis

was performed

using

Student

t-test.

./.

Interactions

between

ethanol

and

15

amphetamines

Results Stereotyped behaviour.Stereotyped behaviour observed after d-amphetamine was very clgfr. The maximum period has ranged from 20 minutes to 90 minutes. (l-2-4 g.kg ) did not modify neither the intensity nor the duration of the behaviour . Toxicity - First ethanol

in

aggregated

mice.-

experiment: (table 2 g.kg

the

toxicity

of

d-amphetamine

of

ethanol

on

toxicity

was

- Second experiment: ethanol was administered aggregated mice remained

d-amphetamine

5 hours

0

IO

10

I

5

8

2

I

I

3

0

0

orally,30 mg.kg

minutes i.p.)

fora 3 g.kg 30 minutes when ethanol

Hyperthermia.The hyperthermia reduced by ethanol (table 2). were manifest 30 minutes after

The its

on

hyprthermia

aggregated

Mean variation post ethanol at 60 min.

0

the

effect-yas (8 mg.kg 4 hours before

induced by hypothermic administration.

-1 own

i.p.) was significantly effects of ethanol

2

and

(“C)

d-amphetamine (8 mg.kg and antihyperthermic

the mst important i.p.). Toxicity in amphetamine.

mortality

induced

by

d-amphetamine

of

rectal temperature post amphetamine at 30 min.

0

+ 3.1

after

Mortality 120 minutes

8

-

0.5

+ 3.7

-

1.3a

+

1.7

a

0

3

-

l.ga

+

1.6

a

I

next

page)

in

mice

1

legend

after

sulfate

2

(see

reduced

mice.

d-amphetamine

dose of ethanol, before d-amphetamine was administered

aggregated Ethaqpl g.kg orally

before

-1

Table

ethanol

in

2 hours

administered

of

strongly

Number of deaths (out of IO) after

(8

Influence

was

1

of

Ethaql g-kg orally

Ethanol

(8 mg.kg-‘)

1). Table

Influence

(2 mg.kg-‘) Ethanol doses stereotyped

7

vhen

16

0. Spreux-Varoquaux

and P. Simon

Ethanol was administerqf orally 30 minutes before the i.p. injection of d-amphetamine (8 mg.kg ). The shown numbers are the mean variations of the rectal temperature ("C) with the initial temperature (10 mice per dose and 20 mice for controls) measured 30 minutes after d-amphetamine administration. Mortality is indicated by the number of deaths per 10 mice, observed 120 minutes after d-amphetamine administration. a: p.

The hypermoti~$ty induced by d-amphetamine was significantly but slightly ethanol (2 g.kg ) (table 3) and was not modified by lower doses (0.5 -

Influence

of ethanol

Table 3 on hypermotility

Etharrrl &kg orally 0 0 2 2

induced

by d-amphetamine

Number of crossed mean -+ s.d.

i.p. 0 3.5 0 3.5

in mice

beams

354 + 147 5552 -+ 278 345 -+ 151 1249 + 178 a

Ethanol was administered orally 15 minutes after the i.p. injection of d-amphetamine. The animals were placed individually in the actimeter 30 minutes after the administration of amphetamine. The indicated results are the mean number of crossed beams observed at 30 minutes (IO mice per dose and 20 mice for controls). a significantly different from amphetamine treated group (p 4 0.01) Discussion In our experimental the effects induced

conditions the oral by amphetamine:

administration

of ethanol

showed a dissociation

of

- a clear reduction of toxicity in aggregated mice, - a partial suppression of hyperthermia in mice (to be compared to the fact that amphetamine (2 mg.kg ) slightly antagonized hypothermia induced by ethanol (Abdallah and Roby, 1975), - and a partial suppression of hypermotility in mice while it was only slightly modified (Breese et al., 1974), and slightly lengthened (Todzy and Becker, 1974) in rats, - a lack of activity they were lengthened

on stereotyped and increased.

behaviour

in rats

while

Todzy and Becker

(1974)

reported

This kind of antagonism is conanon with drugs of sedative patterns,and ethanol shows a profile kindred to anxiolytic one (Spreux-Varoquaux, 1977). Biochemical mechanisms and central structures involved in the activity of these two substances having to be welI.-defined we still remain in descriptions. Two points have nevertheless to be emphasized. On the one hand, some metabolic interactions have been described in the literature: ethanol is thought to inhibit the parahydroxjrlation amphetamine for it has been put forward: - in rats cerebral

an increase of blood concentration (Jonsson et al., 1973; Rech et al., 1976) and concentration (Rech et al., 1976) of amphetamine coupled with a decrease of

of

Interactions

between

ethanol

urinary concentration of the metabolites of amphetamine - in mice: a suppression for 6 hours of the amphetamine On the other receptors since quaux, 1977). Protection pharmacology

(Creaven et al., 1970), metabolism (Iverson et al.,

hand, it may be assumed that ethanol does not affect directly stereotyped behaviour induced by apomorphine is not modified

against except

toxicity in aggregated mice being neuroleptics, we thought interesting

17

and amphetamines

1975).

the dopaminergic (Spreux-Varo-

a rather unfrequent effect in psychoto report this property of ethanol.

References ABDALLAH, A.H. and ROBY, D.M. (1975). Antagonism of depressant activity of ethanol by DH-524; a comparative study with bemegride, doxapram and d-amphetamine (3 8640). Proc. Sot. Exp. Biol. Med. 148: 819-822. ASKEW, B.M. (1962). Hyperpyrexia as a contributory factor in the toxicity of amphetamine to aggregated mice. Br. J. Pharmacol. 19: 245-257. BOISSIER, J.R. and SIMON, P. (1965). ztion de la caf6ine sur la motilit6 spontanle de la souris. Arch. Int. Pharmacodyn. Ther. 158: 212-221. BREESE, G.R., COTT, J.M., COOPER, B.R., -GE, A.J. and LIPTON, M.A. (1974). Antagonism of ethanol narcosis by tryrotropin releasing hormone. Life Sci. 14: 1053-1063. BROWN, D.J., HUGHES, F.W., FORNEY, R.B. and RICHARDS, A.B. (1966x Effects of d-amphetamine and alcohol on attentive motor performance in human subjects. Proc. IVth Int. Conf. Alcohol and Traffic Safety, 215-219. CREAVEN, P.J., BARBEE, T. and ROACH, M.K. (1970). The interaction of ethanol and amphetamine metabolism. J. Pharm. Pharmac. 22: 828-831. FORNEY, R.B., HUGHES, F.W. and GREEBACH, W. (1964). Measurement of attentive motor performance after alcohol. Percept. Mot. Skills 19: 151-154. HUGHES, F.W. and FORNEY, R.B. (1964). Dextr;amphetamine, ethanol and dextro-amphetamine ethanol combinations on performance of human subjects stressed with delayed auditory feedback (D.A.F.). Psychopharmacologia (Berl.) 6: 234-238. IVERSON, F., COLDWELL, B.B., DOWNIE, R.H. and %lITEHOUSE, L.W. (1975). Effect of ethanol on toxicity and metabolism of amphetamine in the mouse. Experientia 31: 679-680. JARBE, T.U.C. (1977). Alcohol discrimination in gerbils: interactions with bemegride, DH-524, amphetamine and 0 9-THC. Arch. Int. Pharmacodyn. Ther. 227: 118-129. JARBE, T.U.C. and OHLIN, G.C.H. (1977). Interactions between alcohol and other drugs on open-field and temperature measurements in gerbils. Arch. Int. Pharmacodyn. Ther. -227: 106117. JONSSON, J. and LEWANDER, T. (1973). Effects of diethyldithiocarbamate and ethanol on the in vivo metabolism and pharmacokinetics of amphetamine in the rat. J. Pharm. Pharmac. -25: 589591. KAPLAN, H.L., FORNEY, R.B., RICHARDS, A.B. and HUGHES, F.W. (1966). d-amphetamine, alcohol Proc. IVth Int. Conf. Alcohol and d-amphetamine alcohol combination on mental performance. and Traffic Safety: 211-214. The effect of ethanol and amphetamine mixtures on LEONARD, B.E. and WISEMAN, B.D. (1970). the activity of rats in a Y-maze. J. Pharm. Pharmac. 22: 967-968. MERLO, A.B., FABIAN, H.E.M., CHEMERINSKI, E. and BILLIETTM. (1976). Effect of d-amphetamine, Pharmacol. Biochem. Behav. 6: 239-242. ethanol and genever on learning in the rat. Interactions between amphetamine and RECH, R.H., VOMACHKA, M.K. and RICKERT, D. (1976). alcohol and their effect on rodent behavior. Ann.N.Y. Acad. Sci. 281~426-440. REIFENSTEIN, E.C. (1941). Amphetamine sulfate-ethyl alcohol antagon?& in the rabbit. J. Lab. Clin. Med. 27: 131-139. SCHECHTER, M.D. (1974). Effect of propranolol, d-amphetamine and caffeine on ethanol as a discriminative cue. Eur. J. Pharmacol. 29: 52-57. SPREUK-VAROQUAUK, 0. (1977). Psychopharmazlogie expgrimentale de 1'6thanol. These Doctorat Phanaacie, Paris. SWINYARD, E.A., CLARK, L.D., MIYAHARA, J.T. and WOLF, H.H. (1961). Studies on the mechanism of amphetamine toxicity in aggregated mice. J. Pharmacol. Exp. Ther. 132: 97-102. TAYLOR, J.D., WILSON, L., NASH, C.W. and CAMERON, D.F. (1964). The effex of ethyl alcohol and amphetamine on performance. Proc. Canad. Fed. Biol. Sot. 1: 36.

'18

.

0. Spreux-Varoquaux

and P. Simon

TODZY, I. and BECKER, W. (1974). Proceedings: alteration of the effects and the metabolism of d-amphetamine by ethanol. Naunyn. Schmiedebergs Arch. Pharmacol. 282: Suppl.: R 99. WILSON, L., TAYLOR, J.D., NASH, C.W. and CAMERON, O.P. (1966). The cozned effects of ethanol and amphetamine sulfate on performance of human subjects. Can. Med. Ass. J. -94: 478-484.

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Facult6 de Msdecine Piti&Salp^etriRre F 75634 Paris Cedex 13