African Journal of Biotechnology Vol. 8 (21), pp.

5979-5984, 2 November, 2009

Available online at http://www.academicjournals.org/AJB
DOI: 10.5897/AJB09.565
ISSN 1684–5315 © 2009 Academic Journals

Full Length Research Paper

Oxytocic and anti-implantation activities of the leaf

extracts of Graptophyllum pictum (Linn.) Griff.
(Acanthaceae)
Stella O. Olagbende-Dada1*, Grace E. Ukpo2, Herbert A. B. Coker2 and Samuel A. Adesina3
1
Department of Pharmacognosy, University of Lagos, Lagos, Nigeria.
2
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Lagos, Lagos, Nigeria.
3
Department of Physiology, College of Medicine, Idi-Araba, University of Lagos, Lagos, Nigeria.
Accepted 24 September, 2009

This study was aimed at evaluating Graptophyllum pictum aqueous extract (GPAE) and ethanol extract
(GPEE) in vitro for oxytocic and in vivo for anti-implantation activities. The oxytocic screening of the
extracts was carried out on the isolated strip of gravid rat uterus in mid pregnancy and was compared
with the activity of an agonist drug, oxytocin. GPEE exhibited oxytocic activity which is comparable to
oxytocin while GPAE was found to reduce the normal contraction of the uterine strip. The anti-
implantation investigation was done using three groups of eight week old virgin female Sprague-Dawley
albino rats (eight rats/group). A selected dose (400 mg/kg) of GPEE was orally administered to a group
of the rats. The same dose of GPAE was similarly administered to another group while the vehicle of
administration (distilled water) was similarly administered to the third group as control. All
administrations started on day one of pregnancy and were given daily for seven days. The rats were
sacrificed on day 10 of pregnancy. Presence of foetus, implantation sites and number of corporal lutea
in the autopsied rats were recorded and used to calculate the percentage anti-implantation effect.
GPEE, GPAE and distilled water have percentage anti-implantation value of 93.8 ± 9.1, 16.8 ± 8.5 and 3.9
± 5.4, respectively. The results support the use of this plant in folkloric medicine as a delivery aid and
also suggest that the plant can be used very early in pregnancy as a contraceptive.

Key words: Anti-implantation, oxytocic, Graptophyllum pictum, contraceptive.

INTRODUCTION

Numerous herbs have been reportedly used historically tives options particularly women living in the rural areas
by women to aid child delivery, stimulate menstrual flow in developing nations with very high population like India,
or reduce fertility (Bodhankar et al., 1974; Farnsworth et China, Africa (Nigeria) and Bangladesh (World population
al., 1975). Modern scientific studies in experimental Data sheet, 2008). Studying the potency and toxicity of
animals have confirmed the effects of some of these local plants that are reputed for birth control in the
herbs in the reproductive system (Prakash et al., 1985; folkloric medicine of these countries may generate
Desta, 1994; Uguru et al., 1998; Badami et al., 2003). greater confidence in and wider acceptance of herbal
Herbal contraceptives offer alternatives for women who contraceptives.
have problems with or lack access to modern contracep- Graptophyllum pictum is commonly called caricature
plant or Joseph’s coat (due to its bicolour which makes it
attractive). Though foreign to Nigeria G. pictum grows
profusely during the rainy season in the country and can
*Correspondent author. E-mail: gbendedada@yahoo.com. Tel: easily be cultivated by vegetative propagation at this
+234 1-8783443 or +2348023635808. period. It is mainly used as ornamental plant to adorn the
5980 Afr. J. Biotechnol.

home gardens in Nigeria. Elsewhere it was reportedly obtained from the animal centre of the College of Medicine, Univer-
used in folkloric medicine as poultice on cuts, wounds sity of Lagos, Nigeria. It was anaesthetized with 20% pentobar-
bitone (Mayer and Baker Nig. PLC) administered intramuscularly at
and all kinds of swellings and for the treatment of ulcer,
0.2 g/kg and the uterus was rapidly removed by midline incision into
abscess, haemorrhoids etc (Perry, 1980; Kasahara and the lower abdominal cavity. The screening was performed by sus-
Mangunkawatjia, 1986). pending 3 pieces of 2 mm longitudinal cut segment of the uterus in
Although some pharmacological studies have been 3 different organ baths, each containing 20 ml of De-Jalon solution
carried out on this plant (Ozaki et al., 1989; Kusumawat maintained at 37°C. The lower end of the muscle was fixed to a
et al., 2002), there is no report of study on its effect in the glass capillary tube with continuous supply of oxygenated gas (95%
O2 and 5% CO2) to keep the muscle alive while the upper end was
reproductive system. Incidental observation (by this suspended by a thread attached to a Grass polygraph (model 7D)
principal author) of safe delivery of a pregnant goat with Force Transducer which transformed the muscle contraction
shortly after ingestion of G. pictum leaves formed the into a proportional electrical signal. This signal was recorded using
basis of this research. Also the reported research findings an electrically driven Grass polygraph recording chart at a speed of
of Elujoba et al. (1985) that some oxytocic agents can as 5 mm/min. Oxytocin was used as the standard drug at 0.1 – 10
i.u/ml and the extracts at 25-80 mg/ml. The oxytocin and the
well serve as anti-implantation agent when administered
extracts (GPAE and GPEE) dissolved in distilled water at known
early in pregnancy formed the basis for the anti-implan- concentrations were dispensed into the 3 organ baths respectively
tation study. using Eppendorf pipettes (Figure 3).

MATERIALS AND METHODS Evaluation of anti-implantation activity

Plant material and preparation of extracts of G. pictum Virgin female rats exhibiting normal oestrus cycle were selected for
this study. They were randomly put into 3 groups of 8 members
The leaves of G. pictum were collected in the early hours of the each. The vaginal smear of each member of the group was daily
morning from a residential area in Isolo a suburb of Lagos state, examined for proestrus. Any member in which this was established
Nigeria, in August 2007. They were identified by Mr. Odewo, was then removed from the others and caged overnight with a
Forestry Research Institute of Nigeria (FRIN), Ibadan, Nigeria. A proven male for mating. The presence of spermatozoa in the
voucher specimen (PCLACAN01) was deposited in the Institute vaginal smear on the following day is taken as evidence of mating
herbarium. The leaves were dried at a temperature of 40°C for 18 and the day is recorded as day 1 of pregnancy. An effective dose of
h in an oven, 250 g of the coarse powder was extracted in acidic 400 mg/kg of GPEE selected according to Elujoba et al., (1985), an
water (pH 4.2) or 70% ethanol (pH 6.0) by heating at 40°C for 30 equal dose of GPAE and distilled water (vehicle of administration)
min. The plant is basic in nature so optimum extraction was achie- were orally administered daily to members of the 3 groups as
ved in an acidic medium. The extracts were filtered through pre- shown in Table 1 for 7 days starting on day 1 of pregnancy. On day
pleated filter paper (Schleicher and Schuell USA, No 560) and 10 of pregnancy, the animals were laparotomised under light ether
concentrated to dryness at 40°C in a rotary evaporator. Both anaesthesia and sterile conditions. Both uterine horns were
extracts were then suspended in water and defatted by partitioning examined for number of implants, aborted implants and corpora
with n-hexane, the aqueous residue from each extract was freeze lutea; which were recorded.
dried and kept in the refrigerator at 4-10°C until needed. The Data were expressed as a mean ± SD (standard deviation).
aqueous extract (GPAE) yielded 75.6 g (30.2%) while the ethanol Statistical analysis was performed with Student's t-test.
extract (GPEE) yielded 52.8 g (21.1%).

Preliminary phytochemical studies

Animals
Phytochemical screening was performed on both GPAE and GPEE
Virgin female and proven male Sprague-Dawley rats (weight, 120- for the presence of secondary metabolites using the following test
130 g) obtained from NAFDAC animal centre (Yaba-Lagos) was methods; alkaloids -, with Mayer’s and Dragendorff’s reagents
used. The animals were kept in a well-ventilated room in the Labo- (Farnsworth, 1966; Harborne, 1998), tannins - with 1% gelatine and
ratory Animal Centre of the College of Medicine, University of NaCl solutions; flavonoids-with the use of Mg and HCl (Silva et al.,
Lagos, Lagos, Nigeria. They received standard rat chow (Pfizer 1993; Houghton and Raman, 1998) and saponin with the ability to
Feeds Nigeria, Plc) and water ad libitum. produce suds and haemolyse red blood cells (Houghton and
Raman, 1998).
Chemicals and reagents

De-Jalons solution (NaCl 9.0 g, 10% KCl-4.2 ml, 1 M CaCl2-0.27 ml, RESULTS AND DISCUSSION
glucose 0.5 g, NaHCO3 0.5 g - all made to 1 liter solution in distilled
water), Oxytocin (Sandoz, Brazil). The alcoholic extract GPEE showed agnostic effect which
was rapid in onset as shown in Figures 2 and 4; but the
Oxytocic screening
aqueous extract GPAE, suppressed the normal uterine
contraction (Figure 1). The preliminary Phytochemical
A female rat in mid-pregnancy (12 days after conception) was screening showed the presence of saponin, tannin and
 Olagbende-Dada et al. 5981

Table 1. Anti-implantation screening for the 400 mg/kg of both alcoholic and aqueous extracts of G. pictum.

Mean wt (g) (F) (C) Total (L) No of Mean % Anti- Mean %

(A) No. of aborted No of
on day 1 of No of conception Corpora Corpora implantation Anti-
Group Implantation sites death
pregnancy foetus (F+A) lutea lutea L-C/L ×100 implantation
0, 2, 0, 0, 0, 0, 0, 0, 0, 2, 0, 0, 8, 9, 7, 8, 100, 78, 100, 100
GPEE (alcoholic extract) 127.4 ± 2.7 7.9 ± 1.0 None 93.8 ± 9.1
1, 0, 0, 0. 0, 0, 1, 0 1, 0, 1, 0 6, 8, 9, 8. 83, 100, 89, 100
6, 6, 8, 5, 1, 0, 0, 3, 7, 6, 8, 8, 8, 8, 9, 10, 12.5, 25, 11, 20,
GPAE (aqueous extract) 125.8 ± 4.1 8.4 ± 0.9 None 16.8 ± 8.5
7, 6, 8, 6. 0, 0, 0, 0 7, 6, 8, 6 9, 8, 8, 7. 22, 25, 0, 14.3
10, 9, 9, 8, 0, 0, 0, 0, 10, 9, 9, 8, 10, 9, 10, 9 0, 0, 10, 11,
Control (distilled water) 128.0 ± 3.0 9.4 ± 0.7 1 3.9 ± 5.4
9, 10, 8, 9. 0, 0, 0, 0 9, 10, 8, 9 9, 10, 8, 10 0, 0, 0, 10

Figure 1. Effect of aqueous extract of G. pictum (GPAE) on the gravid rat uterus.

flavonoids in glycosidic forms in both extracts muscle are used to induce labour or abortion. extract GPAE exhibiting a depress-sant effect on
while the water extract showed the presence of Such drugs include oxytocin, ergometrine and the normal uterine contraction (Figure 1); while
alkaloid-like sub-stance in addition. quinine (Bowman and Rand, 1980). The oxytocic the alcohol extract GPEE exhibited agonistic
Clinically, drugs that contract the uterine smooth screening of G. pictum showed the aqueous effect (Figure 2) which was comparable in magni-
5982 Afr. J. Biotechnol.

Figure 2. Effect of alcohol extract of G. pictum (GPEE) on the gravid rat uterus.

Figure 3. Effect of oxytocin on the gravid rat uterus.

tude with oxytocin. This observed uterus-contracting 0.1 i.u/ml with progressive increase in magnitude to 1.0
action of GPEE was fast in onset and could be totally i.u/ml after which increase in concentration did not
eliminated by washing with the extract-free Dejalon produce further increase in magnitude.
solution. This may suggest the presence of low molecular The progressive decrease at higher concentration with
weight active compound(s) in the extract, which pene- time in the oxytocic action of GPEE may be due to
trated rapidly to its site of action. While GPEE exhibited a metabolic changes in the structure of the active
strong and progressive increase in contraction at initial ingredients. The chemical constituent of this extract were
low concentration up to 20 mg/ml (Figure 4), increase in identified to be mainly glycosides which may quickly be
concentration after this, that is, > 20 mg/ml showed a degraded by hydrolysis, the product of which is usually
progressive decrease in observed contraction. Whereas not as water soluble as the un-degraded parent
the contraction exhibited by oxytocin though not strong at compound.
the lower concentration of < 0.1 i.u/ml was stronger after The in vivo anti-implantation study supported the in
 Olagbende-Dada et al. 5983

Figure 4. Graph of Amplitude of contraction versus dose (aqueous and alcoholic extracts of G.
pictum and oxytocin).

vitro oxytocic screening in that the alcohol extract GPEE ACKNOWLEDGEMENT

exhibited high percentage of anti-implantation (93.85%)
while the aqueous extract GPAE exhibited very little The principal author, Mrs. S.O. Olagbende-Dada, is
(16.8%) when compared with the control (3.9%). grateful to the University of Lagos, for the research grant
The phytochemical screening result showed the two No.VC/PU/6 awarded for the screening of some plants
extracts to contain similar constituents except for the with birth control properties, Prof S.O. Sofola,
prominent presence of alkaloid-like constituent in the Department of Physiology for allowing the use of his
aqueous extract which was not detected in the alcohol laboratory for the oxytocic study and the authority of the
extract. This alkaloid-like constituent may be responsible animal centre, College of Medicine, University of Lagos
for the suppressant effect on the uterine normal for the use of the animal centre for the anti-implantation
contraction and the low anti-implantation activity exhibited studies, and Mr. Peter Ojobor of the Central Research
by the aqueous extract GPAE. Laboratory, College of Medicine, University of Lagos for
The statistical evaluation of this work, as recom- technical assistance.
mended by the W.H.O. (1981) indicated that the results
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