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Phytomedicine Vol. 2 (4), pp.

293-296,1996
© 1996 by Gustav Fischer Verlag, Stuttgart· Jena . New York

Analysis of the Gastrointestinal Relaxing Effect


of the stem extract of Gongronema latifolium

K. S. GAMANIEL and P.A. AKAH

Department of Pharmacology and Toxicology, NationalInstitute for Pharmaceutical Reserarch and Devel-
opment, P.M.B. 21, Abuja, Nigeria.

Summary

Aqueous extract of the stem of Gongronema latifolium was screened for effects on the gas-
trointestinal tract of rats and mice. On the isolated rat intestine, the extract (5-80 ug) evoked re-
producible and concentration-related relaxation which were attenuated by yohimbine and
phenoxybenzamine. Intraperitoneal (i. p.) administration of the extract in mice significantly re-
duced small intestinal transit dose-dependently. This effect was also attenuated by yohimbine and
phenoxybenzamine, but not prazosin and propranolol. Phytochemical analysis revealed the pres-
ence of some bioactive constituents, while the i. p. LDso value of the extract in mice was estab-
lished as 1678.63 ± 78 mg/kg. The results suggest the involvement of the alphaj-adrenoceptors in
the effect of the extract on the intestinal function.

Key words: Gongronema latifolium, gastrointestinal relaxing effect, a- 2 -adrenoceptor involve-


ment.

Introduction and isolated rat intestine in an effort to study the effects on


gastro-intestinal parameters.
Gongronema latifolium, Benth (Asclapiadaceae) is a per-
ennial edible plant with soft and pliable stem. It is widely
used in the West-African subregion for a number of medic- Materials and Methods
inal and nutritional purposes (Dalziel, 1937). In Sierra
Leone a decoction or cold infusion of the pounded stem is Adult Wistar rats (150-200 g) and Swiss albino mice
used for colic and intestinal symptoms usually associated (20-25 g) of either sex maintained at the Animal Facility
with worms (Deighton, 1957). In Ghana, the boiled fruits Centre of the National Institute for Pharmaceutical Re-
are used as laxative. In the Eastern states of Nigeria, the search and Development (NIPRD) Abuja, were used for the
plant locally known as "Utazi" is a popular spice. The experiments. The animals were fed with standard Pfizer
leaves are used to prepare food for mothers that have re- feeds. Food was withheld 12 hours before the experiments,
cently been put to bed, where it is believed to stimulate ap- but there was free access to water.
petite, reduce post-partum contraction and enhance the re-
turn of the menstrual cycle. The use of the leaves in the
Plant materials
treatment of diabetes melitus is now gaining acceptance
among the populace. There are few reports of the phar- The plant materials were collected from the natural hab-
macological profile of this widely used plant, and this pro- itat in Nsukka, Enugu State, Nigeria in June 1994. Botani-
vides a good enough reason for this study. Therefore, in this cal identity was confirmed by Mr. A. Ozioko (Herbarium
study, a preliminary phytochemical screening of the stem Section), Department of Botany, University of Nigeria
extract was carried out. The biological activities of the Nsukka (UNN). A voucher specimen of the plant is depos-
aqueous extract of the stem were evaluated in intact mice ited in both the University of Nigeria, Nsukka and Nation-
294 K. S. Gamaniel and P. A. Akah

al Institute for Pharmaceutical Reserarch and Development nist was tested on a separate preparation, and there were 5
Herbaria. determinations for each experiment.

Preparation of the extract Studies on small intestinal transit


After removing the leaves, the stem was cut into small The effect of the extract on small intestinal transit in mice
pieces, air-dried for five days and milled to a coarse powder. was tested using the charcoal meal method (Akah, 1989; Di
25 g of the powder was marcerated in a percolator with Carlo et al., 1994). Animals were pre-treated with i. p. injec-
250 ml of distilled water. The mixture was allowed to stand tion of the extract (100 and 200 mg/kg). Control animals re-
for 24 hours after which it was filtered and the filtrate ceived equivalent volume of normal saline as vehicle. Char-
freeze-dried to obtain a solid residue (2.4 g) referred to as coal meal, a suspension containing 5% charcoal in 10%
the extract. Appropriate concentrations of the extract were aqueous solution of tragacanth (0.3 ml/mouse), was admin-
made in distilled water for experiments. istered orally to each mouse 30 min after the administration
For all tests two extracts of different origin (batches) of the extract or normal saline. In some experiments yohim-
were prepared. Both extracts showed in a pre-TLC the bine (1 mg/kg), phenoxybenzamine (1 mg/kg), prazosin
same chromatographic pattern. The measured activities of (1 mg/kg) and propranolol (1 mg/kg) was administered sub-
both extracts showed no statistical significant differences. cutaneously 15 min before i. p. administration of the extract.
The animals were sacrificed 30 min after the charcoal meal,
and the small intestine was rapidly removed and layed out
Acute toxicity test
on a white paper for inspection and measurement of the dis-
The acute toxicity (LDso) of the extract was estimated in tance traversed by the charcoal. The length (from the pylor-
30 Swiss albino mice by the i. p. route as described by Mill- us to the ceacum) travelled by the charcoal plug was calcu-
er and Tainter (1944). In brief, the method involved the ad- lated as a percentage of the intestine length. In all the experi-
ministration of 5 different doses of the extract to 5 groups ments there were 5 mice in each group and/or dose level.
of mice (6 mice/group). The number of deaths in each Results were expressed as means ± SEM. The significance
group within 24 hours was recorded. The LDso was esti- of difference between means was tested by the Students t-
mated from the graph of percentage (%) mortality (con- test, and results were regarded as significant when P < 0.05.
verted to probit) against log-dose of the extract - probit 5
being 50%.
Results

Phytochemical tests The preliminary acute toxicity test in mice established the
i. p. LDso of the extract to be 1678.63 ± 78 rug/kg. Phyto-
The freshly prepared extract was subjected to phyto- chemical analysis revealed the presence of alkaloids, carbo-
chemical analysis as described by Trease and Evans (1983). hydrates, tannins, saponins, flavonoids and glycosides (ex-
cept cynogenetic glycosides).
Studies on isolated rat intestine
Effect on the isolated rat intestine
Segments of rat ileum (2 em long) removed from freshly
killed animals were suspended in a 20 ml organ bath con- The extract (5-80 ug) produced a concentration-depend-
taining aerated Tyrode solution maintained at 37°C. The ent and sustained relaxation of the intestine (Tab. 1). It was
composition of the Tyrode solution was (mmollL) NaCI possible to construct three dose-response-curves with the
137; KCI2.7; csci, 1.0; NaHC0 3 12; NaHzP0 4 0.2 and extract within 60 min without remarkable change in sensi-
glucose 5.6. The tension on the tissues was 1 g. The tissues tivity. Acetylcholine (5.5 x lO-SM)-induced contraction and
were equilibrated for 60 min during which the bathing so- noradrenaline (3.0 x 1O-7M)-induced relaxation of the
lution was replaced every 10 min. At the end of the equili- preparation were also recorded for comparison (Fig. 1).
bration period responses were established non-cumulative-
ly for the extract acting for 45 sec. The responses were re-
Effect of antagonists
corded on Ugo Basile Microdynamometer 7050, through
isometric transducer 7004. A 3 min time cycle was found to Prazosin (2.4 x 10-6M) and propranolol (3.4 x 1O-6M)
be adequate. The effect of yohimbine, phenoxybenzamine, did not influence the extract-induced response (Fig.2). Phe-
prazosin and propranolol on the responses of the submaxi- noxybenzamine (3.6 x 1O-7M) attenuated the relaxant ef-
mal concentration of the extract (20llg) was investigated. fect of the extract (Fig.3 a). Yohimbine (3.2 x 10- 7 - 1.3 x
The antagonists were incubated with the tissue 2 min be- 10-6M) caused a dose-dependent block of the relaxant ef-
fore re-establishing responses to the extract. Each antago- fect of the extract (Fig.3 b).
Analysis of th e Gas tro inte stinal Rela xin g Effect of the stern extract of Gongronema latifolium 295

Table 1. Effect of increasing concentration of the extract on the


isolated rat small intestine (n =5).
Concentration of Extract Relaxation (g)
(ug) Mean ± SEM
5 0.65 ± 0.013
to 0.96 ± 0.23
20 1.05 ± 0.033
40 1.17±0.0 19 5 10 20 40 80u9 5 10 20 /,0 8O U9
80 1.18 ± 0.068 extract extra ct

Fig. 1. Concentration-dependent responses of the extract


(5-80 ug) on isolated rat small intestine. Acetylcholine (ACh 5.5 x
Table 2. Effect of phenoxybenzamine (PBZ ), yohimbine, prazosin 10-8M) and noradrenaline (NA 3.0 x to-7M) induced responses
and propran olol on the inhibitory activity of the extract on small are shown.
intestinal transit. Number of animals is shown in parentheses.
*P < 0.05 versus vehicle contro l, ,.,. P < 0.05 versus extract
200 mg/kg. NS = normal saline.
Treatment Length of Intestine % inhibition
(mg/kg) travelled (em) ± SEM
Vehicle (NS) 36.31 ± 2,4 (5)
Extract 100 22.31 ± 1,4" (5) 37.18 *
Extract 200 13.14 ± 2.3" (5) 63.12*
Extract 200 + PZB I 32.26 ± 4.2 (5) 11.15**
Extract 200 +
yohimbine 1 34.61 ± 1.1 (5) 4.68 " "
Extract 200 + prazosin 1 14.04 ± 2.1 (5) 61.33
Extract 200 + Fig.2 . Effect of prazosin (PR 2,4 x 10-6M) and propranolol (PRO
propranol ol 1 12,43 ± 3.0 (5) 65.76 3,4 x to-6M ) on relaxant effect of the extract (E 20 /lg).-

Effect on small intestinal transit


In the control mice, the charcoal meal mo ved 65.61 ±
4. 63% of th e tot al len gth of th e intestine . The extract 100
and 200 mg/k g admi nistered i. p. significa ntly (P < 0.05) re-
du ced in a dose-relat ed manner th e mo vement of th e char-
Fig.3. (a) Showing the relaxant effect of the extract (E 20/lg) an-
coal meal (Tab le 2) . These mo vements we re significa ntly tagonised by phenoxybenzamine (PBZ 3.6 x 1O-7M) and (b) con-
different (P < 0.05) fro m the co ntrol. Phenoxybenzam ine centration-dependent block by yohimbine (YB) of the relaxant ef-
(1 mg/kg) and pr a zosin (1 mg/kg), w hich did not influence fect of the extract (E 20 ug),
per se the mo vem ent o f th e cha rcoal meal , red uced sign ifi-
ca ntly (P < 0.05) th e inhibito ry effect of th e extrac t
(200 mg/k g). O n th e o ther hand th e effec t of the extract o n 1994), and th e ro le of alphaj -adrcnoce pto rs was prop osed
th e inte stinal tr an sit was not infl uence d by pr azosin (Di Carlo et aI., 1994). Stimulat ion of alpha--ad renocc p-
(l mg/kg) and pro pr an olol (1 mg/kg). tors ha s been reported to induce gut relaxat ion a nd inhib i-
tion of sm all intestinal transit (Ruwa rt et al., 1980; H su ,
1982). In th e present report, th e extract induced relaxation
Discussion of th e intes tine a nd inhibition o f the sma ll intestina l tran sit
was attenuat ed effectively by th e relatively specific alpha .-
The results of th is stu dy clearly ind icat e th at the adminis- adr enoceptor antagonists, yohi mb ine a nd phenoxy benza -
tr ation of th e stem aq ueo us extract of G . latifolium pro- mine (Berla n et al., 1992; Bylund, 1992; H offman and Lef-
duced potent relaxatio n of th e gas trointestinal tract an d in- kowitz, 1990), but not by prazosin and propranolol, an al-
hibition of sma ll int estinal transit . The reversibility, repro - ph a.-adrenoceptor a nd B-adrenoceptor blockers respect ive-
d ucibility and dose-related effects a re rem arkable. There ly (Fo rd et al., 1994; Mekean and MacDo nald, 1994).
are many bioacti ve co nstit uents pr esent in the extra ct and These dat a tend to suggest t he involvement of alpha .-
hence, at present, it is not certain, whi ch of them is/ar e re- adrenoceptors, and it is likely, th ou gh not proven, th at fla -
spo nsible for th e o bserved effect s. H owever, some repo rts vonoids co ntri bute in pa rt to th e gas trointes tinal re laxi ng
ha ve sho wn th at flavonoids inh ibit intesti na l motibil ity in a effect of G . latifolium. Sim ilar results have been reported
dose-related manner (M eli et aI., 1990; Di Carlo et aI., by Visw an athan et a1. (1984) and Di Ca rlo et a1. (1994). AI-
296 K. S. Gamaniel and P. A. Akah

though we have proferred evidence implicating alpha.- Deighton, E C.: Vernacular botanical vocabulary for Sierra Leone.
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