M Abubakar, U Suleiman, A Frank, A Ukwuani
aqueous extract, d. metel, rats. acetylcholine, scopolamine
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.
The hallucinogenic effect of aqueous seed extract of D.
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.
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: firstname.lastname@example.org
Materials and Method
Collection of Plant Material
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).
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.
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;
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.
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 was determined according to method of Bessey
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.
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.
Data were expressed as mean
Gas chromatography mass spectrophotometry
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
Effect of aqueous seed extract of on heart and respiratory rate.
Table 2: Shows the effect is the aqueous seed extract of
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
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
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
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
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
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
The findings of our research clearly validate the action of