British Homoeopathic Journal July 1997, Vol. 86, pp. 139-141

Suppression of alpha adrenergic agonist-induced catalepsy in mice by potentized Agaricus musearius SOUVIK GHOSH, MSC, S.P. SINHA BABU, MSC, PHD, N.C. SUKUL, MSC, PHD

Abstract Agaricus muscarius 30c, a potentized homoeopathic drug prepared by successive dilution with 90% ethanol followed by sonication in 30 steps, suppressed catalepsy induced by alpha adrenergic agonists in Swiss albino mice. Agaricus produced anticataleptic effect when it was administered orally and no such effect when administered intraperitoneally. The alpha 1 agonist phenylephrine and alpha 2 agonist clonidine were administered intraperitoneally to mice at a dose of 2 mg/kg and 1 mg/kg, respectively. Mice were pretreated orally with Agaricus muscarius 30c. The action of Agaricus is thought to be mediated through ororeceptors.

KEYWORDS: Alpha agonists; Phenylephrine; Clonidine; Catalepsy; Homoeopathic potency; Agaricus muscarius; Oral route; Anticataleptic; Ultrasonic; Ororeceptor.

Introduction Potentized Agaricus muscarius significantly suppresses haloperidol-induced neuroleptic catalepsy in albino mice. This anticataleptic effect of Agaricus 30c is dose-dependent, l, 2 The effects of various antipsychotic drugs are described as a 'neuroleptic syndrome' char- acterized by psychomotor slowing, emotional calming and affective indifference. These drugs reduce aggressiveness, impulsiveness, initiative and anxiety without a change in consc iousness or in te l lec tua l facul t ies . Neuroleptics belong mainly to 5 chemical classes of which bu tyrophenone is one. Haloperidol is a substituted piperidine com- pound of butyrophenone. Neuroleptic drugs are mainly used to treat schizophrenia. Given in higher doses, neuroleptics produce catalepsy) Catalepsy is a state of profound movement inhi- bition, characterized by an inability to correct awkward postures without failure of righting reflex. 4 It is a central nervous system phe- nomenon involving several neurotransmitter systems. Cholinergic agonists such as pilo- carpine and dopaminergic antagonists such as haloperidol promote catalepsy. The situation can be reversed by a cholinergic antagonist such as atropine sulphate and a dopaminergic agonist such as apomorphine. 5-7

Catalepsy can also be induced by alpha adrenergic agonists such as phenylephrine and clonidine and reversed by their specific antagonists such as phenoxybenzamine and yohimbine. 8 Agaricus is mentioned as one of

the anticataleptic drugs in Kent's Repertory of Homoeopathic Materia Medica. 9 It is therefore interesting to see whether Agaricus could counter catalepsy induced by neuro- transmitter drugs other than haloperidol. In this study the effect of potentized Agaricus on mouse catalepsy induced by alpha adren- ergic agonists was determined. Agaricus was given both orally and intraperitoneally to see which of the 2 routes was effective.

Materials and methods Subjects Male albino mice weighing 25-30 g were used in these experiments. They were housed in groups of 5 in conventional cages with bedding. All the animals were kept in an animal house with natural light and a room temperature of 28 + 2~ They were tested for catalepsy at least 2 weeks after shipment.

Drugs tested The mother tincture of Agaricus muscarius was obta ined f rom Boer icke and Tafel Company, Philadelphia, USA. The tincture was diluted 1 : 100 with 90% ethanol in our laboratory. The mixture was sonicated at 20 Hz for 30 seconds using an ul t rasonic hom o g en i ze r (Labsonic 2000, Braun, Germany) to produce the 1st centesimal potency Agaricus lc. Subsequent potencies up to 30th were produced by adding to 1 part of the preceding potency 99 parts of ethanol and sonicating the mixture in the same way.

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Observation has shown that sonication produces a more effect ive potency than manual agitation, l~ Sucrose pilules were wetted with freshly prepared Agaricus 30c in a glass vial at the rate of 0.1 ml of the drug per 3.1 g of pilules. The control comprised sucrose pilules wetted in 90% ethanol diluted and sonicated in the same manner without using the mother tincture of Agaricus at the beginning. Both the drug and the control were dissolved in dis- tilled water at the rate of 10 pilules (112 rag) per ml of distilled water and given orally to mice at the rate of 0.05 ml/mouse by glass pipette. In another set of experiments the drug and the control solutions were injected into mice intraperitoneally.

The alpha 1 agonist phenylephrine and the alpha 2 agonist clonidine, obtained f rom Sigma Chemical Co., USA, were mixed with sterile distilled water and injected intraperi- toneally into mice at doses of 2 mg and 1 mg/kg body weight respectively. The salt form of each drug was mixed with water and given to mice in a volume dose of 10 ml/kg. Control groups were given 0.9% saline i.p. Agaricus 30c was administered 4 h before adrenergic agonists and catalepsy tests were performed 0.5 h after the administration of agonists.

250

200

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Drug Combinations

FIGURE 1. Reduction in duration of catalepsy induced by alpha 1 agonist phenylephrine and alpha 2 agonist clonidine given intraperitoneally to albino mice at 2 mg/kg and 1 mg/kg respectively resulting from pretreatrnent with Agaricus muscarius 30c by oral route. Agaricus administered i.p. did not reduce duration of catalepsy induced by alpha agonists.

Tests for catalepsy Catalepsy tests were performed in a quiet room by the inclined floor method 8 at 17-18 hrs. A smooth bottomed enamel tray (32 x 30 cm) was kept at a 70 ~ incline with bedding material at the bottom rim. Pieces of sandpaper (2 x 1 cm) were glued alternately in a chequer board pat- tern in 3 rows near the upper end of the tray. A paper-free border of 2.5 cm surrounded the che- quer board at the sides and the top. A mouse was held by its tail and placed horizontally on the sandpaper mosaic. The t ime a mouse remained immobile on the strips was taken as the catalepsy duration. A cut-off time of 300 seconds was maintained. 5 groups of mice, each numbering 10, were tested for catalepsy for each drug, drug combination or control. The data for every 5 groups of mice for each treat- ment/control were pooled and a single median duration of catalepsy obtained. A single median thus represented the individual catalepsy dura- tion of 50 mice. The experimenter was blind to the test conditions.

Results The median duration in seconds of catalepsy for each treatment/control is shown in Figure 1. Agaricus 30c alone produced a very mild catalepsy in mice (Figure 1). Both alpha ago- nists, phenylephrine and clonidine produced significant cataleptic effect as compared to the control (p < 0.005, Kruskal-Wallis test). Post- hoc t-tests for drug pairs showed that catalepsy duration was significantly lower with Agaricus 30c (oral) plus phenylephrine or clonidine than with either adrenergic agonist alone (p < 0.01). The duration of cata lepsy produced by Agaricus 30c (i.p.) plus either phenylephrine or clonidine did not differ significantly from that produced by either agonist alone (Figure 1). Mice treated with alpha agonists alone and Agaricus 30c (i.p.) plus either agonist were visi- bly immobi le in their cages just before catalepsy tests. However, mice treated with Agaricus 30c (oral) plus either agonist were actively mobile like control animals just before catalepsy test.

Discussion Catalepsy induced by neuroleptics such as haloperidol has been attributed to blockade of striatal dopamine receptors.11-~4 Brainstem adrenergic neurons project rostrally and caudal- ly to a variety of movement control systems of

Volume 86, July 1997

the brain. Alpha adrenergic agonist molecules bind to the specific receptors in the area and may induce ca ta lepsy . Since p o t e n t i z e d Agaricus reversed catalepsy mediated either by doparninergic or noradrenergic systems in different areas of the brain, there may be a common nucleus influencing both systems. Agaricus may act on the common nucleus in the brain and thus reduce catalepsy in both cases. The DA receptor blocker haloperidol in large doses reduces catalepsy and increases noradrenaline metabolism in the rat brain. 15 This suggests a possible interaction between the dopaminergic and noradrenergic systems in relation to catalepsy. Since the drug is in a highly dilute state (10 60) and is effective only by the oral and not the intraperitoneal route, its action is mediated by oral receptors. The main finding in this study is that potentized Agaricus in ultra-high dilutions could reverse adrenergic drug-induced catalepsy.

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4 Baez LA, Burt DK, Granneman J, Shanklin C. Dopaminergic antagonism and catalepsy in the developing rat. Eur J Pharmacol 1979; 54: 15-20.

5 Klemm WR. Experimental Catalepsy: Influences

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8 Sukul NC, Cherian L, Klemm WR. Alpha nora- drenergic agonists promote catalepsy in the mouse. Pharmacol Biochem Behav 1988; 31: 87-91.

9 Kent JT. Repertory of the Homoeopathic Materia Medica 1877 p. 1347. Calcutta: Seth Dey 1961.

10 Sukul NC. Mechanical agitation, the main factor in increasing the efficacy of Agaricus muscarius, a homoeopathic drug. Environ EcoI 1992; 10: 7-10.

11 Carlsson A, Lindquist M. Effect of chlorpro- mazine or haloperidol on formation of 3-methoxytyramine and normetanephrine in mouse brain. Acta pharmac toxic 1963; 20: 140-4.

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15Toru M, Takashima M. Haloperidol in large doses reduces the cataleptic response and increases noradrenaline metabolism in the brain of the rat. Neuropharmacology 1985; 24:231-6.

Address f o r correspondence Professor N.C. Sukul Department of Zoology Visva-Bharati University Santiniketan-731235 West Bengal India