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  • The Internet Journal of Pharmacology
  • Volume 9
  • Number 1

Original Article

Hallucinogenic Effects Of Aqueous Seeds Extract Of Datura Metel In Rats.

M Abubakar, U Suleiman, A Frank, A Ukwuani

Keywords

aqueous extract, d. metel, rats. acetylcholine, scopolamine

Citation

M Abubakar, U Suleiman, A Frank, A Ukwuani. Hallucinogenic Effects Of Aqueous Seeds Extract Of Datura Metel In Rats.. The Internet Journal of Pharmacology. 2009 Volume 9 Number 1.

Abstract

The hallucinogenic effect of aqueous seed extract of D. metel was evaluated. The ethanolic extract was subjected to Gas Chromatography Mass Spectrophotometry and the result shows the presence of an alkaloid scopolamine. Male wister rats were divided into four groups and were orally administered with aqueous seed extract of 0.0, 0.4, 0.6 and 0.8mg/kg body weight respectively. The treated groups exhibited some behavioral changes such as restlessness, aggressiveness, agitation and disorientation. The effect of the extract on the food and water intake shows a significant decrease (p<.05) in the 0.6 and 0.8mg/kg extract treated groups as compared with control. However, the heart rate increased significantly (p<.05) in 0.6 and 0.8mg/kg treated groups while the respiratory rate increased in the 0.8mg/kg treated group as compared with control respectively. A decrease of vitamin A, C and E was observed at all dose level particularly in 0.8mg/kg extract treated group (p<.05) at 3.27± 0.1, 4.08± 0.3 and 0.28± 0.04µg./dl respectively. The hallucinogenic effect observed may be due to the presence of the alkaloid scopolamine.

 

Introduction

The use of botanicals in their natural unprocessed form undoubtedly began when the first intelligent animal noticed that certain plants altered particular body function (Katzung, 2004). Hallucinogenic plants have been use as mind altering agents since the beginning of the recorded history. Besides being their use for therapeutic purposes, these agents are also use inform of abuse by criminals because of the fact that the agents in question produce a transient psychotic state to the user called “hallucination”; which is a sensory experience of something that does not exits out side mind. Hallucinations are defined also as “an apparent perception of an external object when no such object is present. It is to be differentiated from illusions in which real perceptions are misinterpreted (Hinsie and Campbell, 1970). It may involve distorted sensory perception so that things look differently from what they are (Marek, 1998). Consequences of the use of hallucinogenic plants for abuse include criminal tendencies men, prostitution in women and other social unrest.

Datura metel is one of the most interesting plants with hallucinogenic properties. The plant is used for the treatment of asthma, depression, motion sickness and analgesic (Bonde, 2001). The watery extract of the leaves is applied around the eyes for the treatment of cataract (Conklin and Marie, 1976). The seeds were also use some time for criminal purposes. The criminal intoxicate themselves with boiled seed before perpetrating their acts.

The widespread, uncontrolled use of these plants has made it necessary to apply scientific knowledge in order to ascertain the nature of these plants. The present study was therefore undertaken to investigate the possible active compound responsible for causing hallucination to the users.

Corresponding Author: A. N. Ukwuani, Department of Biochemistry, Faculty of Science, Kebbi state university and technology, Aliero, Nigeria. Email: pinknnenna@yahoo.com

Materials and Method

Collection of Plant Material

D. metel seeds were procured from different damp places within the Sokoto metropolis; the plant was identified and verified at Usmanu Danfodiyo University Sokoto, Herbarium (Botany Unit, Department of Biological Sciences). The seeds obtained were open-air-dried under the shade, pulverized into a moderately coarse powder (using pestle and mortar).

Solvent Extraction and Fractionation

Fractionation of the extract was done by activity guided fractionation using ethanol-water (1:1) and different (methanol, chloroform and petroleum ether) organic solvents. The powdered plant extract (50g) was extracted with 200mL of ethanol-water (1:1) for 24hrs and filtered. The filtrate was partitioned in 100mL methanol, the methanol extract was then removed, and the aqueous filtrate (ethanol-water) was further partitioned with 100mL petroleum ether. The petroleum ether extract was then removed and then finally the aqueous extract (water-ethanol) was partitioned with chloroform and then the chloroform extract was removed (Safowara., 1982).

Aqueous Extraction

A 200g of powered seeds were soaked in 2liters of distilled water and allowed to stand for 48 hours at room temperature. It was filtered and the filtrate was evaporated at 450C to obtain residue, the residue yield 15.8g. The residues were reconstituted in sterilized distilled water (Safowara, 1982).

Gas Chromatography Mass Spectrophotmetry

The four fraction of the solvent were analyzed in GC-MS using Acq method. One microlitre (1µL) of the sample was injected into the column with rate flow of 0.5µl/min. The initial temperature of the column is 800C, the rate of heating is 100C/min. The inlet temperature is 2500C, while the final temp is 2800C, the total running time is 30minutes.

Experimental Animals

Twenty male albino rats weighing between 239.7 ± 47.3g were obtained from the Department of Pharmacology, Aminu Kano Teaching hospital and were then transported in a well ventilated cage to Usmanu Danfodiyo University, Sokoto, and kept in animal house. They were allowed free access to water and feed (marsh). They were left in the environment for 2 weeks to acclimatize

Extract Administration

The animals were divided into four groups of five rats each. Animals in group 2, 3, and 4 were given oral administration of 0.4, 0.6 and 0.8mg/kg of the aqueous extract respectively. The group 1 received distilled water. The procedure is shown below;

Figure 1

The extract was administered for 2 days and after the extract administration each day, the behaviour of the animals were monitored. The respiratory and heart rate were also measured. The respiratory rate was measured by counting the number of breathing per minute, and the mean was obtained from triplicate counts. Similarly, the heart rate was measured by feeling the pulse at the wrist and then the number of pulse per minutes was recorded. The mean was obtained by triplicate values ( Anika and Shetty, 1982). The rates of food and water consumption were also monitored by measuring the food and water intake, before and after the following morning. The animals were then allowed for 72 hours in which each day their behaviour and their physical body parameters were observed and monitored.

Brain analysis

The animals were anesthetized with chloroform vapour and dissected. The brains were then removed and placed in isolation buffer solution (phosphate buffer pH 7.0). Then the brain was homogenized using pistil and mortar, then centrifuge at 4000rpm for 10minutes. The supernatant was removed and then subjected to the analysis of vitamin A, C and E.

Vitamin A

Vitamin A was determined according to method of Bessey etal., (1946), which is based on the addition of ethanol to break up the complex and permit vitamin A to portion in for the heptane. The nearly colourless retinal is measured spectrophotometrically at 326nm.

Vitamin C

Vitamin C was measured according to lowrey method (1957). Which is based on oxidation of ascorbic acid to dehydroascorbic acid by tricholoracetic acid. The dehydroascorbic acid in acidic solution react with 2,4-dinitrophenyl hydrazine forming corresponding hydrazone. The hydrazone when treated with sulfuric acid develop orange-red colour which is measured sphectrophotometrically at 520nm.

Vitamin E

The micro method for estimation of vitamin E was adopted which is based on. The reduction of ferric ions to ferrous ions which form red colour with α- α-dipyridyl.

Statistics

Data were expressed as mean + SEM and subjected to analysis of variance (ANOVA) using two way method for accessing statistical significance.

Results

Gas chromatography mass spectrophotometry

Figure 2
Fig. 1.0 shows that, the GC-MS recorded the ethanolic seed extract of the to contain an alkaloid scopolamine.

Effect of aqueous seed extract of on behaviour and physical parameters on rats.

All the treated rats exhibit some behavioural changes such as restlessness, agitation, aggressiveness and disorientation, pupils dilation and dryness of the mouth was also observed.

Effect of aqueous seed extract of on food and water intake.

Take 1: shows the effect of the aqueous seed extract of D.metel on food and water intake. There was significant difference (P<0.05) in group 4 (0.8mg/kg ) as compared with control.

Effect of aqueous seed extract of on heart and respiratory rate.

Table 2: Shows the effect is the aqueous seed extract of D.metel on rate of respiration and heart rate. The results shows a significant increase (P<0.05) in groups 3 and 4 as compared with control.

Effect of aqueous extract of on vitamin A, C and E in brain.

Table 3: Shows the effect of the extract on vitamin A,C, and E in the brain of the rats. The result shows a significant decrease (P<0.05) with increase in dose of the D.metel extract.

Figure 3
TABLE 1: EFFECT OF EXTRACT ON FOOD AND WATER INTAKE.

Figure 4
TABLE 2: EFFECT OF EXTRACT ON RESPIRATORY RATE AND HEART RATE.

Figure 5
TABLE 3: EFFECT OF EXTRACT ON VITAMIN A, C AND E, IN BRAIN.

Discussion

Hallucinogens are compound that caused hallucination. Hallucination is sensory experience of something that does not exist outside mind. It may involve distorted sensory perception so that things look sound, smell, or feel differently from the way they are. Although typically associated with psychiatric disorder, the hallucinatory experience has a wide range of etiologies that may include but not limited to the following: neurological insult, seizure and sleep disorders, drug reaction, substance abuse, grief, stress as well as metabolic, endocrine and infectious diseases (Hinsie and Campbell, 1970). The compound cause hallucination by mimicking or other wise affect neuro effectors junctions. Such agent exerts their influences in several ways; interference with the synthesis or release of the transmitter, interact with receptor and causes the destruction, dispersal or dissipation of transmitter (Wilhem, 1964).

GCMS result obtained recorded, that the ethanolic extract shows the presence of scopolamine. This scopolamine is structurally analogues to a neuro transmitter acetylcholine, and therefore inhibits the effect of the neuro transmitter. Acetylcholine is found in both peripheral nervous system (PNS) and in the central nervous system (CNS) as a neuro modulator (katzung, 2003).

Scopolamine competitively inhibit the acetylcholine at muscarinic acetylcholine receptors found in both peripheral and central nervous system (Foye et al.,1995). The decrease in locomotor activity immediately after extract administration as observed may be due to the inhibition of acetylcholine at M3 receptors of the ending vagus nerves of the muscle. Normally acetylcholine is released when an exited neurons diffuses a few micrometers across the synaptic cleft or neuromuscular junction to the postsynaptic neuron or myocyte, where it interacts with it receptor and triggers electrical excitation (depolarization) of the receiving cell, depolarization of the muscle fibre triggers muscle contraction (Michael and David, 2005).

The emergence of pupil’s dilatation treated groups shows the peripheral action of the extract even in lower dose which may be the paralysis of the oculomotor nerve ending or its myoneural junction.The dryness of the mouth of the treated animals suggest that, the extract have effect on submaxillary gland that control the secretion of saliva. Since muscarinic receptors when bound to acetylcholine in submaxillary gland stimulate gastric acid secretion, salivation and lacrimation (Foye et al., 1995). Therefore, the absence of saliva indicates the competitive activity of the extract on the gland.

The restlessness and hyperactivity exhibited by the treated animals serve as a marker for the action of the extract on central nervous system which comprises the brain and the spinal cord. Acetylcholine has effect on excitability of central nervous system (CNS), its presence causes a slow depolarization by blocking a tonically active K+ current which increase neuronal excitability (Katzung,2004) while in the brain muscarinic receptor (M5) is found in substantial nigra where it regulate dopamine release at terminals within the striatum (Foye et al., 1995). This could be probably the reason for Hallucination effects of the D. metel by affecting the dopamine release.

The food and water intake significantly (P<0.05) decrease with increase in the dose of the extract throughout the experiment as compared with control, indicating that, the extract suppress apatite. From the result the effect of extract on heart rate shows significance (P<0.05) increase with increase in the dose of the extract. Acetylcholine induces decreased contraction in cardiac muscle fibers by binding to its M2 receptors found in cardiac muscle (Foye et al., 1995). But due to the presence of scopolamine, it inhibits the activity of the acetylcholine at the receptor and hence result in rapid heart beat.

The rate of respiration is also significantly (P<0.05) increase as compared with control. The increase in respiration is due to the Broncho dilation that occur when the bind to the M3 receptor on air way smooth muscles. In the airways acetylcholine is released from efferent ending of the vagus nerves and bind to the M3 receptor there by mediating boroncho constriction (katzun,2004). The competitive inhibition of scopolamine to the acetylcholine results in broncho dilation hence increase in respiration.

The effect of extract on vitamin A, C, and E shows that the D. metel extract significantly (P<0.05) decrease the levels of these vitamins in the brain suggesting that the extract depletes these vitamins in the brain. Vitamin A, C and E are called antioxidants vitamin, they scavenge the oxygen free radicals which cause lipid peroxidation which causes delayed reversible demylinazation of white matter in the central nervous system, and can lead to edema and focal areas of necrosis within the brain (Gorman et al., 2003). Therefore, the potential of causing brain damage by the extract of D. metel has been significantly elucidated by the result of this study.

The findings of our research clearly validate the action of D.metel on central nervous system (CNS) and pheriparal nervous system (PNS) and this provide the evidence to support the use of such plant for medicinal, social or even rituals in the past.

References

1. Anika, S.M. and S.N Shetty,(1982). Laboratory Manual of Pharmacology and Toxicology. First Edition., First Dimention Publishers, London pp 23.
2. Bonde, K.(2001). The genus Datura; From Research subject to Powerful Hallucinogen. Journal of Ethanobotanical Leaflet Vol29 335-336
3. Conklin, and E. Marie, (1976). Genetic and Biochemical Aspect of the development of Datura. Monographs in the Developmental Biology new York.
4. Foye, W.O., lemke T.L., will Arms, D .A, and W. Wilkins, (1995). Principle of medicinal chemistry. Fourth Edition, Oxford University Press, London.
5. Gorman, D. Drewry A. huang Y.T.and C. James,(2003). The Clinical Toxicology of Carbon monoxide. Journal of Toxicology (1) pp 25-8
6. Hinsie, L and Campbell, R (1970). Psychiatric Dictionary. Fourth edition. Oxford university press. Newyork.
7. Katzung, B.G., (2004). Basic and Clinical pharmacology. 9th Edition, Mcgraw Hill Medical, New York.
8. Macheal, L. andL.N David (2004). Lehninger Principles of biochemistry 4th Edition, Free Man and Co, New York.
9. Marek, G.J. (1998). Serotonin 5-HT2A receptor enhance asynchronous excitatory transmission in pyramidal cells (layer v) of pre frontal cortex. Journal of society for neurosciences vol. 24, 365-7
10. Safowara, A. (1983). Medicinal plant and traditional medicine in Africa, 1st edition John W. Chicester U.K.
11. Wilhem, K. and O. Renner, (2005). Phamacokinatics and Phamacodynamics in clinical use of scopolamine Journal of therapeutic drugs Monitoring. Vol 27 pp 655-665.

Author Information

M. G. Abubakar
Department of Biochemistry, Faculty of Science, Usmanu Danfodiyo University

U. Z. Suleiman
Department of Biochemistry, Faculty of Science, Usmanu Danfodiyo University

A.S. Frank
Department of Biochemistry, Faculty of Science, Usmanu Danfodiyo University , Sokoto , Nigeria .

A. N. Ukwuani
Department of Biochemistry, Faculty of Science, Kebbi State University of Science and Technology

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