| http://www.aygrt.isrj.net/UploadedData/1397.pdf Executive Editor :- |
Ashok Yakkaldevi258/34 ,Raviwar Peth ,Solapur-413005, Mahashtra,India.Contact On : +91-9595 359 435, +91 9096938215 |
THE EFFECTS OF AEGLEMARMELOSON MOTOR
CO-ORDINATION IN WISTAR RATSAND IT'S COMPARISON
WITH DIAZEPAM
HIMANI ,R. C. VERMA , PRATAP SHANKAR , SHARADLEVE AND R. K. DIXIT
Junior Resident. Pharmacology. K.G.M.U. Lucknow.
Lecturer Pharmacology. R.I.M.S. Saifai, Etawah.
Junior Resident. Pharmacology. K.G.M.U. Lucknow.
Junior Resident. Pharmacology. K.G.M.U. Lucknow.
Professor Pharmacology. K.G.M.U. Lucknow.
Abstract:
Present study was done to see the effect of Aeglemarmelos on motor coordination in wistar rats using rota rod method. Result of present study showed positive
motor co-ordination effectof Aeglemarmelos.Animals receiving A. marmelosextract
(100 mg/kg, 200 mg/kg) showed significant increase in the time spent on revolving rod
when compared to control, thus demonstrating the positive effect on motor coordination.
KEY-WORDS:
Aeglemarmelos, motor co-ordination, rota rod.
INTRODUCTION:
Aeglemarmelos, commonly known as a bael, is one of the gifts of nature to mankind. Numerous
pharmacological studies have been conducted on different parts of Aeglemarmelos. This plant is having
great potential to cure the diseases like diabetes, hyperlipidemia, peptic ulcer, diarrhoea, dysentery, cancers
etc. It has also shown its effect as cardio protective, anti bacterial, anti fungal, radio protective, anti pyretic,
analgesic, antioxidant, hepatoprotective, and many more.
However, there are only few studies of Aeglemarmelos pertaining to central nervous system
activities. Hence in the present study, Neuropsychopharmacological effects of Aeglemarmelos (Bael)
(effect on motor function) were studied in wistar rats.
The patterns of voluntary activity are planned within the brain, and the commands are sent to the
muscles primarily via the corticospinal and corticobulbar systems. Posture is continually adjusted not only
before but also during movement by descending brain stem pathways and peripheral afferents. Movement
is smoothed and coordinated by the medial and intermediate portions of the cerebellum (spinocerebellum)
and its connections. The basal ganglia and the lateral portions of the cerebellum (cerebrocerebellum) are
part of a feedback circuit to the premotor and motor cortex that is concerned with planning and organizing
voluntary movement. Alpha motor neuron (lower motor neuron) are the final common pathway.
The harmonious functioning of body parts that involve movement, including gross motor
movement, fine motor movement, and motor planning. Central Control of Motor Function governed by the
Cerebral (association) cortex (generates the impulse to act, the design and the planning of movement);
Basal ganglia and thalamus (provide the gross motor “programs); Motor cortex, brain stem and spinal cord
(interact to maintain posture and movement execution); Cerebellum (maintains equilibrium and smooth
coordination of movement (refines movement)).
Plants are the very important for the human, because they posses several activeconstituents which
are the precursor for synthesis of many useful drug (Dhankhar and Ruhil, 2011). Their effectiveness, low
cost and comparative freedom from serious toxic effects make these medicines not only popular but also an
acceptable mode of treating diseases even in modern times. Due to the various unavoidable adverse effects
of available allopathic medicines, management of various diseases without any untoward side effects is still
a challenge for modern medical science. So several herbal plants having various bioactive phytochemicals
have been explored, possessing several activities with minimal adverse effects.Bael is one of the gifts of
nature to mankind.
MATERIALS AND METHODS
The study was conducted in the Department of Pharmacology and Therapeutics, King George's
Medical University, Lucknow (Erstwhile ChhatrapatiShahujiMaharaj Medical University). Ethical
clearance was obtained from the Institutional Animal Ethics Committee before conducting the study.
Experimental Animals & Rearing Conditions
Adult healthy Male Wistar rats weighing 160-200 gm had been used in study. Animals had been
obtained from CPCSEA-certified animal house [Indian Institute of Toxicology Research, Lucknow
(IITR)]. They were allowed to access normal rat pellet diet and water ad libitum and were kept in
Institutional animal house under temperature controlled environment [25 ± 2°C] with 12 hours' light and
dark cycle. The animals were housed for two weeksprior to the experiments to acclimatize to new
environment.
The maintenance of the animals and the experimental procedures were in accordance with the
guiding principles of Institutional Animal Ethics committee and the 'Guide for the Care and Use of
Laboratory Animals', National Research Council, 1996 (Latest revision in 2011). The guidelines of
Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Govt. of
India were followed.
Dosage Forms, Doses and Sources of the Drugs
Following drugs were used in this study.
(1)Test Drugs - Bael (Aeglemarmelos)- Drug (extract form) was dissolved in normal saline and
administered orally with the help of feeding cannula in a doses of 100 mg/ kgbw and 200 mg/ kgbw
(Shankharananth, 2007). It was purchased from market (Aeglemarmelos extract, Himalaya Drug
Company).
(2)Standard drugs – Diazepam:Dose 2 mg/kgbwi.p. (Sujith et al., 2011; Kumar et al.,2011). Tablets were
purchased from government authorized medical store.
Experimental Protocol
The present study had been designed to evaluate Neuropsychopharmacological effects of
Aeglemarmelos(Bael) that includes effect on motor function in male wistar rats.
Animal Groups
A total number of 4 Male Wistar rats were included in the study. They were kept in Institutional
Animal House under standard conditions. All the animals received normal rat pellet diet and water ad
libitum. All the animals were allowed to get acclimatised to the new environment for period of
2 weeks.Rats
were randomly divided into 4 groups, each group containing 6 rats:Group 1 to 4 were used to evaluate
analgesic action of Aeglemarmelos and its effect on motor coordination. These actions were compared with
tramadol and diazepam respectively.(2 weeks after the evaluation of analgesic action of Aeglemarmelos,
we again randomly divided 24 animals into 4 groups to study its effect on Motor co-ordination).
Neuropsychopharmacological Evaluation
Adult male Albino Wistar rats weighing between 160- 200 gms were used. All the animals allowed
toget to acclimatize to the new environment for period of 2 weeks. They were provided with normal rat
pellet diet & water ad libitum.Following validated behavioural models of rodents were used to assess the
neuropsychopharmacological effects of Aeglemarmelous extract.
Assessment of Motor function in rats using rotarod apparatus
Grouping
Albino Male Wistar rats weighing between 160-200 gms were randomly divided into 4 groups,
each group containing 6 rats.
Group 1: Rats were administered distilled water p.o (1 ml)
Group 2: Rats were administered aeglemarmelous extract (100 mg/kg) p.o.
Group 3: Rats were administered aeglemarmelous extract (200 mg/kg) p.o.
Group 4: Rats were administered standard drug Diazepam (2 mg/kg) i.p.
PROCEDURE
Motor function test was carried out using rotarod apparatus. The ability of a rodent to maintain
balance and keep pace with a rotating rod has been used with varying degrees of success over the years to
assess motor function. The rotarod apparatus was placed in a height above the table top in order to
discourage the animals from jumping off the roller.The rats were acclimatized to the revolving drums, and
they were habituated to handling to ameliorate any stress during testing. Animals remaining on Rota-Rod
(16 rpm) 2 min or more in three successive trails were selected 1 day before the actual day of testing (Saha et
al., 2010). The selected animals then randomly were divided into four groups (n=6). The animals that held
onto the rotating rod for 2 min were placed again on the same rotating rod on the following day (actual day
of testing). The mean of three training runs served as a control performance time (Basal reading). The
rotarod performance time was measured at 30, 60, 90 and 120 min after administration of drugs and vehicle
(Yee et al., 2009). The fall off time from the rotating rod was noted (Naggar et al., 2003; Perez et al., 1998).
ASSESSMENT
The duration of stay on the rotating rod was noted. Mean of the each group was calculated and
used for the comparison of control and drug treated groups.
TECHNIQUE OFINTRAPERITONEALINJECTION
To perform an intraperitoneal injection, the rat must be well restrained so that it cannot move
during the procedure. This avoids traumatizing the organs once the needle entered the abdomen. The animal
was restrained in such a way that the abdomen was exposed. An imaginary line was drawn across the
abdomen just above the knees. The 26 G needle was inserted at a point on this line on the animal's right side
and close to the midline after disinfecting the injection site. The needle was inserted into the abdomen at an
angle of 30 degree and not more than a depth of half centimetre. The small intestines (on the right side) are
less likely to be punctured by the needle. Inserting the needle too far caudally or laterally from the insertion
point would risk making an injection into the rear leg which would injure the muscle tissue.Before injecting
drugs, a gentle aspiration was done to make sure that the needle had not penetrated a blood vessel, the
intestines, or the urinary bladder. In case any fluid was aspirated, the contaminated solution was discarded
and the procedure repeated with a new syringe and needle. After injection, the site was again cleaned and
animal was observed.
ORALDRUG ADMINISTRATION
The desired doses of drugs were calculated on the basis of body weight and dissolved in normal
saline just prior to administration. It was then loaded in a syringe fitted with a 16 G ball tipped feeding
needle. The rat was restrained using a towel in the left hand. The head of the rat were restrained using the
thumb and index finger whereas the tail of the rat was restrained between the ring and little finger. While the
rat was lying on the left palm, the ball-tipped feeding needle was introduced from the side of mouth in to the
pharynx and then let in to the esophagus when the animal was in the act of swallowing. The drug was then
injected in to the esophagus of routs.
STATISTICALANALYSIS
A one-sample Kolmogorov-Smirnov test was used to investigate whether the variables were
normally distributed. The one way analysis of variance (ANOVA) was used to assess the comparability of
the groups assigned to the treatment groups. Independent t test/Tuke's pairwise comparison was used to
compare the different parameters like immobility time, transfer latency, open arm activity, number of visits,
reaction time and duration of stay on rotarod between respective treatment groups. Differences in treatment
effects within groups and between the treatment and control groups were tested by a multivariate analysis of
variance repeated-measures design with treatment type as a between-subject factor (2 groups) and
treatment effect (baseline compared with follow-ups) as a within-subject factor. A significant P value for
the treatment effect indicated a change over time in the combined values of the groups and was further
investigated by using a paired t test for each individual group. Between group differences in treatment
effect were indicated by significant interactions between treatment effect and treatment type. The percent
change from baseline to follow-ups was also calculated for each group. Statistical significance was based
on a two-tailed Pvalue < 0.05.
OBSERVATIONS AND RESULTS
Assessment of MotorFunction Activity
The effect of the different treatment group on motor function activity have been summarized in
Table-1 and also shown graphically in Fig. 1 The motor coordination activity was insignificantly increased
from 0 min to 30, 60, 90 and 120 min (p>0.05) in Group 1. However, the motor coordination activity was
significantly increased from 0 min (141.00±4.29) to 30 (190.83±4.75, p<0.05), 60 (250.33±4.71, p<0.01),
90 (230.17±5.67p<0.01) and 120 min (200.83±7.03, p<0.01) in group 2. Almost similar observation was
found for Group 3. The motor coordination activity was significantly decreased from 0 min (149.67±6.21)
to 30 (20.33±0.81, p<0.01), 60 (21.33±0.82, p<0.01), 90 (22.83±0.75, p<0.01) and 120 min (22.83±0.75,
p<0.01) for Group 4.
The motor coordination activity was insignificantly different between groups at 0 min (p>0.05).
However, motor coordination activity was significantly different between Group 1 and Group 2 (p<0.01), 3
(p<0.01) and 4 (p<0.01) at 30 min. All the groups were significantly different to each other at 30 min, 60
min, 90 min and 120 min (Table-2).
Golden Research Thoughts • Volume 2 Issue 7 • Jan 2013 4
Table 1: Effect of Aeglemarmelous extract onMotor function(ROTAROD):
The motor coordination activity was insignificantly different between groups at 0 min (p>0.05).
However, motor coordination activity was significantly different between Group 1 and Group 2 (p<0.01), 3
(p<0.01) and 4 (p<0.01) at 30 min. All the groups were significantly different to each other at 30 min, 60
min, 90 min and 120 min (Table-2)
DISCUSSION
Due to the various side effects of allopathic drugs used for treatment of these diseases, there is
continuous search for alternative treatment. So it is prudent to look for options which are efficacious &
safer. Indigenous system of medicine including natural herbs are time tested way of treatment. Herbal
medicines emphasize the prevention of diseases, rejuvenation of our body systems, maintain balance and
harmony and extend the life span (Mahe et al., 1978). Medicinal herbs are indispensible part of traditional
medicine practiced all over the world due to easy access, low cost and ancestral experience. Number of
plants have been being used for management of mental illness. Some of them are as follows:
Atotal number of 24 male Wistar rats were included in the study. Rats were randomly divided into
4 groups, each group containing 6 rats. All the animals were allowed to get acclimatised to the new
environment for period of 2 weeks. Group 1to 4 were used to assess the effect of Aeglemarmelos on pain
and motor function. Motor function test was carried out using rotarod apparatus. The ability of a rodent to
maintain balance and keep pace with a rotating rod has been used with varying degrees of success over the
years to assess motor function.
The standard drug diazepam showed marked changes in motor function. Animals treated with
diazepam showed significant reduction in the time spent on the revolving rod when compared to control. In
contrast, A. marmelosextract (100 mg/kg, 200 mg/kg) significantly increased the time spent by the animals
on revolving rod when compared to control, thus demonstrating the positive effect on motor coordination.(Table 1,2,3).
Hence, due to the presence of a number of phytoconstituents including flavonoids, quercetin,
tannic acid, phenols, eugenol, marmesinin, ascorbic acid, skimmianine and saponin etc. or any other
mechanisms.Aeglemarmelos possesses improved effect on motor co-ordination.Aeglemarmeloscan be a
safe and effective indigenous drug for the treatment of number of psychiatric disorders.However, a more
extensive study is necessary to determine the exact mechanism of action of the extracts and its active
compound(s).
CONCLUSION
The present study was designed to evaluate the neuropsychopharmacological effects (effect on
motor function) of Aeglemarmelos extract. These effects were compared with standard drugs of their class
(Diazepam).Result of present study showed positive motor co-ordination effectof Aeglemarmelos.
Animals receiving A. marmelosextract (100 mg/kg, 200 mg/kg) showed significant increase in the time
spent on revolving rod when compared to control, thus demonstrating the positive effect on motor coordination.
Present study [an experimental study to evaluate the neuropsychopharmacological effect (effect
on motor function) of Aeglemarmelos extract] shows that Aeglemarmelospossessesimproved motor coordination in rats. These findings are in favour of using A. marmelosasimproved motor co-ordinationdrug.
However the results from present study have limitations in the form of short duration of study, only one or
two selected models and less number of animals. If A. marmelos passes through the positive results in
animal study, clinical studies may be planned in future.
REFERENCES:
1.Control and Supervision of Experiments on Animals (CPCSEA), Govt. of India
2.Dhankhar S, Ruhil S, et al., (2011).Aeglemarmeloscorrea: A potential source of phytomedicine. JMPR
5(9):1497-1507.
3.Goel RK, Maiti RN, Manickam M, Ray AB (1997).Antiulcer activity of naturally occurring
pyranocumarin and isocoumarins and their effect on prostanoid synthesis using human colonic mucosa.
Indian J ExpBiol35:1080-83.
4.Guide for the Care and Use of Laboratory Animals
5.Gupta V, Bansal P, Niazi J, Kaur G (2010).Anti-anxiety activity of Citrus paradise var. star ruby
extracts.IJPTR 2(3):1655-1657.
6.Jagetia GC, Venkatesh P (2005). Radioprotection by oral administration of Aeglemarmelos (L.) correain
vivo.JEnviron PatholToxicolOncol 24:315-332.
7.Kamalakkanan N, Prince PS (2007). Hypoglycemic effect of water extracts of Aeglemarmelosfruits in
Streptozotocin diabetic rats. J Ethnopharmacol 87: 207.
8.Kitamura Y, Araki H, Nagatani T, Takao K, Shibata K, Gomita Y (2004).Influence of Imipramine on the
duration of immobility in chronic forced swim stressed rats.Acta Med Okayama 58(6):271-274.
9.Kumar S, Joseph L, George M, Kaur L, Bharti V (2011). Skeletal muscle relaxant activity of methanolic
extract of Rumexnepalensis in albino rats. J Chem Pharm Res 3(3):725-728.
10.Kumar V(2006). Potential plants for CNS disorders: an overview.Phytother Res20(12):1023-1035.
11.Mahe M, Driessche JV, Girre L (1978). Pharmacological properties of several indigenous plants on the
nervous system. Plant Med Phytother12:248-258.
12.Mukund H, Gawade SP (2011). Evaluation of effects of photooxidizedVespa orientalis venom on
memory and learning in rats. J Venom Anim Toxins Incl Trop Dis17(4):422-429.
13.Naggar TB, Gomez SMP, Carretero ME, Villar AM (2003).Neuropharmacological activity of Nigella
sativaL. extracts. JEthnopharmacol 88:63-68.
14.National Research Council, 1996 (Latest revision in 2011)
15.Parichha S (2004). Bael (AegleMarmelos) nature's most natural medicinal fruit. Orissa Rev16-17.
16.Parmar C,Kaushal MK (1982). Wild fruits of the Sub-Himalayan region. New Delhi: Kalyani
Publishers. pp 1-5.
17.Patel PK, Sahu J (2012).Aeglemarmelos:A review on its medicinal properties.International
JPharmaceutical Phytopharmacological Res1(5):332-341.
18.Perez RM, Perez JA, Garcia LM, Sossa H (1998).Neuropharmacological activity of
Solanumnigrumfruit. J Ethnopharmacology 62:43-48.
19.Purohit SS, Vyas SP (2004). In: Aeglemarmelos Correa ex Roxb(Bael), Medicinal plant cultivation- A
scientific approach.Agrobios Jodhpur 280-285.
20.Raffa RB, Friderichs E, Reimann W, Shank RP, Codd EE, Vaught JL (1992). Opioid and nonopioid
components independently contribute to the mechanism of action of tramadol, an 'atypical' opioid
analgesic. J PharmacolExpTher 260(1):275-85.
21.Rajadurai M, Prince PS (2005). Comparative effects of Aeglemarmelosextract and alpha-tocopherol on
serum lipids, Lipid peroxides and cardiac enzyme levels in rats with isoproterenol-induced myocardial
infarction. Singapore Med J 46:78.
22.Rajput MS, Sinha S, Mathur V, Agrawal P (2011). Herbal Antidepressants. International
JPharmaceutical Frontier Res 1(1):159-169.
23.Roy SK,Khurdiya DS (1995). Other subtropical fruit. In Salunkhe, DK and Kadam SS (eds.), Handbook
of Fruit Science and Technology: Production, Composition, Storage and Processing. New York: Marcel
Dekker.
24.Sabu MC, Kuttan R (2004). Antidiabetic activity of Aeglemarmelosand its relationship with its
antioxidant properties. Ind J PhysiPharmacol 48:81-88.
25.Saha S, Srikanth L, Das S (2010). Effect of Amlodipine on the psychomotor dysfunction induced by
carbamazepine in rats.Int J PreclinicPharmaceut Res 1(1)25-30.
26.Sairam K, Dorababu M, Goel RK, Bhattacharya SK(2002). Antidepressant activity of standardized
extract of Bacopamonniera in experimental models of depression in rats. Phytomedicine9:207-211.
27.Sakakibara H, Yoshino S, Kawai Y, Terao J (2008). Antidepressant-like effect of onion (Allium cepa L.)
powder in a rat behavioral model of depression. BiosciBiotechnolBiochem72:94-100.
28.Shankarnanth V, Balakrishnan N, Suresh D, Sureshpandian G, Edwin E,Sheeja E (2007). Analgesic
activity of methanol extract of Aeglemarmelos leaves.Fitoterapia78(3):258-259.
29.Sharma PC, Bhatia V, Bansal N (2007). Areview on Baeltree. Natural Product Radiance6(2):
30.Singanan V, Singanan M, Begum H (2007). The hepatoprotective effect of bael leaves (Aeglemarmelos)
in alcohol induced liver injury in albino rats.Internat J SciTechnol 2(2):83-92.
31.Sujith K, Suba V, Ronald DC (2011). Neuropharmacological profile of ethanolic extract of
Anacycluspyrethrumin albino wistar rats. Inter J PharmacSci Res 2(8):2109-2114.
32.Venkatesan D, KarrunakarnCM, Kumar SS (2009). Identification of phytochemical constituents of
Aeglemarmelosresponsible for antimicrobial activity against selected pathogenic organisms.
Ethnobotanical Leaflets 13:1362- 1372.
33.Kim YS, Perk HJ, Kim K,Moon DE, Lee IHJ (2009). The effects of Ginkgobilobaextract EGb 761 on
mechanical and cold allodynia in a Rat model of neuropathic pain. AnesthesiaAnalgesia. Intern Anesthesia
Res Soc 108(6):1958-1963.
34.Yu ZF, Kong LD, Chen Y(2002). Antidepressant activity of aqueous extracts of Curcuma longa in mice.
J Ethnopharmacol 83:161-165.
