PHYTOTHERAPY RESEARCH

Phytother. Res. 13, 549–554 (1999)

Cardiovascular Effects of an Extract from the

Roots of a Shrub Elaeophorbia drupifera

A. E. Eno and O. I. Owo

Department of Physiology, College of Medical Sciences, University of Calabar, Calabar, Nigeria.

A crude extract was prepared from the roots of E. drupifera. Lethality studies in mice showed a dose-
mortality relationship with an LD50 of 145 mg/kg mice i.p. The extract (2–260 mg/kg. i.v.) was tested in
graded doses on the blood pressure and heart rate of urethane anaesthetized rats. The results showed
that the extract decreased both the blood pressure and heart rate in a dose-dependent manner. The max-
imum decrease in blood pressure (control, 78.3  6.5 mmHg) and heart rate (control, 120.2  5.5 beats/
min) produced by the extract was about 46.2% and 41.7% (% control), respectively.
Blocking the beta adrenoceptors with propranolol (0.5 mg/kg. i.v.) did not prevent the action of the extract
on both the blood pressure and heart rate, suggesting that the extract was acting at a different site. This view
was supported by the observation that the extract significantly depressed the increase in blood pressure and
heart rate caused by bilateral occlusion of the common carotid artery. Also, the extract was found to prolong
ACh-induced hypotension in the rats. In animals pretreated with atropine sulphate (0.2 mg/kg. i.v), the
extract was less effective in depressing the blood pressure. However, this atropine antagonism was
surmounted by raising the concentration of the extract.
Finally, in vitro studies using isolated arterial strips revealed that the extract also had a relaxant effect on
vascular smooth muscle. This relaxant effect was dose-dependent and was attenuated and/or abolished by
phentolamine (0.5 mg/mL). Also, the extract relaxed aortic strips precontracted with noradrenaline
(1  10ÿ7 mol Lÿ1) but failed to relax strips precontracted with KCl (50 mmol/L).
We conclude that the crude extract from the roots of E. drupifera probably contains acetylcholine-like
agent(s) which interferes with the cholinergic mechanism, as well as catecholamine-like agent(s) exhibiting
mainly alpha-adrenoceptor activity. Copyright # 1999 John Wiley & Sons, Ltd.
Keywords: Elaeophorbia drupifera; roots; blood pressure; heart rate; depression.

INTRODUCTION
Little literature is available on the medicinal uses of the
plant Elaeophorbia drupifera (Thonn.) Stapf; although it
is listed among the plants that heal (Ampofo, 1977). This
local herb is used by traditional herbalists for the
treatment of various ailments. The leaves, stem (bark),
roots or the latex are used depending on the type of
ailment. The fruit is succulent (Kinghorn and Evans,
1975) and the leaf is used as a filaricide and for guinea
worm infestation (Comley, 1990). Recently, the leaf has
been reported to contain hypoglycaemic agent(s) (Eno
and Itam, 1996), and agent(s) that stimulate autonomic
cholinoceptors (Eno and Itam, 1998). According to some
herbalists, the roots and even the leaves are used as
diuretics, antihypertensives, and could relieve muscle
pains and aches resulting from minor spasms or spasticity
when applied topically to the skin or taken orally
(personal communication).
Our main objective for this study was to investigate the
possible effects of E. drupifera roots on the cardiovas-
* Correspondence to: Dr A. E. Eno, Department of Physiology, College of
Medical Sciences, University of Calabar, P M B 1115, Calabar, Cross River
State, Nigeria.
E-mail: nkin@unical.anpa.net.ng
CCC 0951–418X/99/070549–06 $17.50
Copyright # 1999 John Wiley & Sons, Ltd.
cular system. This could provide a scientific basis for its
antihypertensive property, as claimed by many traditional
healers.
MATERIALS AND METHODS
Preparation of crude extract. Fresh roots of E.
drupifera were collected. The roots were first washed
free of soil and debris. Wash water was blotted off and
the roots ground to a paste. A quantity of the ground
sample (50 g) was weighed and soxhlet-extracted with
150 mL distilled water at 100 °C for 8–10 h. Where larger
ground samples (500–1000 g) were used, extraction was
done under reflux with an appropriate volume of water.
The extract was slowly evaporated to dryness in vacuo at
40 °C using a rotary evaporator. A starting sample of 50 g
of fresh material gave a mean yield of 2.35  0.6 g
(SD) of extract (n = 8). Weighed samples of the extract
were then used to make up test solutions ranging in
concentration between 5 and 200 mg/mL (expressed in
terms of the weight of extract).
Acute toxicity test. Male white albino mice (20–25 g)
were randomly assigned to 12 groups of 10 animals per
Received 9 April 1998
Revised 11 September 1998
Accepted 20 October 1998
550 A. E. ENO AND O. I. OWO

group. Each group was injected intraperitoneally with was added cumulatively into the bath at 10 min interval
one of the following: 2.5, 5, 10, 20, 40, 80, 100, 120, 140, which allowed each dose to reach a steady level before
160, 180, 200 mg/kg of the crude extract. The maximum the addition of the subsequent dose. In another set (n = 6)
volume injected was 0.2 mL. The groups were returned to of aortic strips, phentolamine (1.5 mg/mL) was used to
their home cages and given free access to food and water. block the alpha adrenoceptors before the application of
The mortality in each cage was assessed 24 h after the extract (2–8 mg/mL).
administration of the extract. The percentage mortalities
were converted to probits and plotted against the log10 of Statistical analysis. Regression lines with confidence
the dose of the extract. Regression lines were fitted by the limits were calculated for the linear portions of log
method of least squares and confidence limits for the concentration-response curves. The significance of dif-
LD50 values were calculated by the method of Litchfield ferences in slopes was used as a measure of parallelism of
and Wilcoxon (1949). the two lines. Log concentration limits at 50% of the
maximum response were used in the analysis of the
Measurement of blood pressure and heart rate. significance of concentration differences as described by
Normal male white Wistar rats (250–300 g) were Birmingham et al. (1970). Maximum responses were
anaesthetized with 25% urethane at a dose of 6 mL/kg compared by paired Student’s t-test.
body weight. The trachea was intubated and the femoral
vein and carotid artery cannulated (Portex cannulae, Reference Drugs. Phentolamine, propranolol and acet-
external diameter 1.02 mm, internal diameter 0.75 mm). ylcholine chloride were all obtained from Sigma.
The cannulation of the femoral vein and carotid artery Potassium chloride, noradrenaline tartrate and atropine
were for drug administration and blood pressure record- sulphate were from the British Drug House (BDH).
ings, respectively. At the same time, the heart rate was
monitored on a different channel of the same recording
machine (Washington, Model 400 MD/2C). The pressure
transducer (Washington, P. T. 400) was used for blood RESULTS
pressure measurements. The rectal temperature of the rat
was maintained at 37 °  1 °C by means of a rat table Toxicity Test
heater.
The crude extract was injected (i.v.) as a 0.2 mL bolus Lethality studies showed a dose-mortality relationship
at eight different increasing doses to construct a dose- which was apparently sigmoidal. A plot of probit values
response curve. The doses (2, 4, 8, 16, 32, 64, 130, and (% mortality) versus log-dose of extract gave a straight
260 mg/kg) were injected at an interval of 5 min, giving
cumulative doses of 2, 6, 24, 300, 62, 126, 256 and
516 mg/kg extract. The effect of a dose was calculated by
averaging the last minute of blood pressure and heart rate
recording preceding the following dose. In preliminary
experiments, a single dose of the extract produced an
effect lasting for more than 1 h. In another group of rats,
the dose-response experiment was repeated but the
animals received the extract 2 min after atropine
(0.2 mg/kg i.v.) pretreatment.
Blood pressure responses were measured as change in
mean arterial pressure (MAP mmHg), from the pretreated
level. Attempts were made using standard pharmacolo-
gical agents to elucidate the possible mechanism of
action of the crude extract.

Isolated tissue experiments. Rabbits (2–3kg) were

killed either by cervical dislocation or by a single blow
to the head. The abdominal aortae were removed and cut
into helical strips (3–4 cm long). The strips were mounted
vertically in a 25 mL organ bath containing Krebs
bicarbonates solution of the following composition:
(mM concentration) NaCl, 120; KCl, 4.8; CaCl, 1.2;
MgSO4
7H2O, 1.3; KH2PO4, 1.2: NaHCO3, 25.2; and
glucose 5.8; pH 7.4; bubbled with a gas mixture of 95%
O2 and 5% CO2, and maintained at 37 °  1 °C.
A resting tension of 2 g was applied to each strip of the
mounted tissue. Tension changes evoked by the extract
were recorded on a polygraph (Grass Model 7D) via an Figure 1. Dose-effect relationship. The curves show blood
isometric transducer (FT. 03). After allowing a 30 min pressure changes (mmHg) following cumulative administra-
equilibration period, dose-response experiments (n = 6) tion of E. drupifera root extract (2±516 mg/kg i.v.) to a group of
rats (*), and to another group (*) that received atropine
were carried out under baseline tension. Later, the rings (0.2 mg/kg i.v.) pretreatment. Blood pressure is expressed as
were made to contract with the addition of noradrenaline the relative change from the value obtained before the i.v.
(10ÿ7 mol/L) or potassium chloride (50 mmol/L). After injection of the extract. Data are the mean value  SEM (n = 8
the contractions had stabilized, the extract (2–8 mg/mL) per group, p < 0.01).

Copyright # 1999 John Wiley & Sons, Ltd. Phytother. Res. 13, 549–554 (1999)
 CARDIOVASCULAR EFFECTS OF ELAEOPHORBIA DRUPIFERA 551

Table 1. Effects of E. drupifera root extract on the rate of heart beat following drug
pretreatment
Heart rate (beats/min)
Control Drugs (mg/kg i.v.) Extract (10 mg/kg i.v.)
121.3.  4.6 Propranolol (0.5) 113.2  8.3a 106.7  6.4a
23.1  6.8 Carotid occlusion 137.4  7.1a 118.6  5.9b
118.6  8.2 Acetylcholine (0.1) 109.3  6.8 98.3  7.7b
The agents were administered before the injection of the extract (at steady state level).
a
p < 0.01 compared with the matching controls.
b
p < 0.05 compared with the drug treated group.
Data shown are mean  SEM, n = 5, and the concentration of the drugs (in
parentheses).

line. From the straight line graph, the LD50 value of the
extract was extrapolated. This value was about 145.4 mg
dry wt, of extract per kg mice (mg/kg mice i.p.). No
animal died earlier than 3 h. However, the high dose
recipients (140–200 mg/kg) became immobile about
40 min postinjection. They were cold to touch with a
mean rectal temperature of about 24.2  8.5 (SEM,
n = 5), suggesting severe hypothermia.
Effects of crude extract on blood pressure (BP) and
heart rate
The control mean arterial pressure (MAP) in the urethane
anaesthetized rats was 78.3  6.5 mmHg (SEM, n = 8)
before the administration of E. drupifera root extract.
Slow intravenous injection of the extract decreased the
BP of rats in a concentration-dependent fashion. The
extract-induced a concentration-dependent decrease in
BP as shown in Fig. 1 (ED50 = 60.9  3.4 mg/kg). The
maximum decrease in BP (41.3  7.5 mmHg) was
achieved with the extract at a cumulative dose of
516 mg/kg. This represents about 52.6% (% control)
Figure 2. The effects of E. drupifera roots extract (2±516 mg/kg
i.v.) on the rate of heart beat measured simultaneously with
the blood pressure. The group of rats injected with the extract
only is shown by the dotted column while the group
pretreated with atropine (0.2 mg/kg i.v.) is shown by the
hatched column. The control heart rate is the open column.
Data are the mean  SEM (n = 8 per group).
‡
‡‡
p < 0.05 vs control group.
p < 0.05 vs atropinized group.
Copyright # 1999 John Wiley & Sons, Ltd.
depression of blood pressure by the extract. The effect of
the extract on the rat BP was quick in onset and the
recovery time increased with increasing concentration of
the extract.
The control heart rate (HR), measured simultaneously
with the BP was about 120.2  5.5 (SEM, n = 8) beats/
min before the injection of the extract. Following the
administration of the extract (2–516 mg/kg), the HR
decreased dose-dependently (Fig. 2). The crude extract
(515 mg/kg) produced the maximum decrease in HR
(69.6  7.2 beats/min) which represents about 41.7% (%
control) decrease.
Further studies were carried out to elucidate the
possible mechanisms by which E. drupifera root extract
elicited the observed hypotensive effect. In these studies,
the crude extract (10 mg/kg. i.v.) aggravated the hypoten-
sion (10.4%, of controls) caused by propranolol
(0.5 mg/kg. i.v.). The extract produced about
13.6%  5.4% (SEM, n = 5, p < 0.01) further decrease
in BP when compared with that caused by propranolol
(Fig. 3a). Furthermore, the extract (10 mg/kg i.v) almost
abolished the reflex response (increased BP and increased
heart rate) to bilateral occlusion of the common carotid
artery (Perera et al., 1985). This blocking effect persisted
for about 1 1/2 h. The carotid occlusion (Fig. 3b) raised
the BP from the control level (78.3 mmHg) to about
92.5  4.6 mmHg (about 17.5% increase). The extract
(10 mg/kg i.v) reduced the occlusion-induced increase in
BP to about 72.7  6.1 mmHg. This figure represents
about 21.4% depression of BP by the extract, from the
raised level. Finally, the extract (10 mg/kg. i.v) was also
found to prolong the effect of acetylcholine (0.1 mg/kg.
i.v) on blood pressure. Acetylcholine decreased the BP
from the control level to about 73.8  7.4 mmHg.
Injection of the extract (10 mg/kg i.v.) produced a further
fall in BP to about 56.3  6.5 mmHg (about 30.1%
maximum decrease from the control level) (Fig. 3c).
Such an enhancement may also contribute to the
observed hypotensive activity of the extract.
The heart rate responses corresponding to these drug
treatments are summarized in Table 1, and were all in
good agreement with the BP results.
The results above raised a strong suspicion that a
cholinergic mechanism might be involved in the action of
the extract. Consequently, a similar dose-response curve
experiment was repeated in another group of rats (control
MAP = 76.8  7.6 mmHg, n = 8). In this group, the
animals were pretreated with atropine sulphate
(0.2 mg/kg i.v.) to block the muscarinic cholinoceptors,
before injecting the extract (2–516 mg/kg i.v., cumulative
dose range) as described in the methods section. In
Phytother. Res. 13, 549–554 (1999)
552 A. E. ENO AND O. I. OWO

Figure 3. Typical mechanical records showing the effects of the extract on the blood
pressure of rats following pretreatment with propranolol (Prop) (a), bilateral occlusion
of the carotid artery (occlusion) (b) and acetylcholine (ACh) (c).

atropine - treated animals, the extract-induced decrease in

BP was reversed. The dose-response curve
(ED50 = 48.9 mg/kg) was shifted to the left and the
atropine-antagonism appeared to be surmounted with
higher doses of the extract. The maximum decrease in BP
(40.6  6.3 mmHg) of rats pretreated with atropine
before the extract was not significantly different from
the matching controls (Fig. 1).
DISCUSSION
The crude extract from E. drupifera roots appears to be
relatively non-toxic considering its LD50 value in mice.
Although the cause of death in the high dose recipient
mice is not clear, cardiotoxicity may not be ruled out, as

Effects of extract on isolated aortic strips

The effects of E. drupifera root extract on vasodilation

was assayed in vitro using isolated arterial strips. At
concentrations of 2–250 mg/mL bath solution, the extract
consistently relaxed the arterial strips (Fig. 4). The
magnitude of the relaxations were dose-dependent. The
onset of relaxations was fast, but reached the maximum
level much more slowly (5–10 s). Once the arterial strips
relaxed, the tension was maintained for more than 1 h in
the presence of the extract. However, recovery (return to
baseline tension) could be achieved much earlier by
washing the tissue preparation with Krebs bicarbonate
solution.
Some pharmacological agents were used to influence
the relaxant effect of the extract on the aotic tissue (Fig.
5a–e). Cumulative addition of the extract (2, 4, 8, mg/mL)
to the bath solution caused dose-dependent relaxations of
the strips (Fig. 5a) as well as arterial strips precontracted
with noradrenaline (1  10ÿ7) (Fig. 5b) but failed to relax
strips precontracted with potassium chloride (Fig. 5c).
Furthermore, the in vitro vascular effect of the extract Figure 4. Dose-response curve showing the effects of graded
(6 mg/mL) was antagonized and/or abolished by phento- dose of E. drupifera root extract (2±50 mg/mL) on the resting
tension of isolated arterial strips. The relaxations were
lamine (0.5 mg/mL) (Fig. 5d). This blocking effect of expressed as the % decrease in tension development taking
phentolamine was also observed in strips precontracted the resting tension of the strips as 100%. Data are the
with noradrenaline (1  10ÿ7) (Fig. 5e). mean  SEM, n = 6; p < 0.01 vs resting tension.
Copyright # 1999 John Wiley & Sons, Ltd. Phytother. Res. 13, 549–554 (1999)
 CARDIOVASCULAR EFFECTS OF ELAEOPHORBIA DRUPIFERA
Figure 5. Mechanical records of the isolated arterial strips
showing (a) consistent relaxation of the muscle from its
resting tension by increasing doses of the extract, and when
precontracted with (b) noradrenaline (NA), and (c) potassium
chloride (KCI). Also shown is the phentolamine (phen)
antagonism of (d) extract-induced relaxation of the strips,
and (e) strips precontracted with noradrenaline (1  10ÿ7 M).
severe hypotension could reach the level at which blood
flow to vital organs (heart, brain and kidneys) is impaired,
this effect resulting in death (Tarazi and Gifford, 1979).
One important experimental observation about the
general activity of this extract was its fast onset of
action. This may indicate a low molecular weight
compound present in the extract, which may penetrate
rapidly to its site of action (Aldeen et al., 1981).
Alternatively, this could merely reflect the high concen-
tration of the active compound present in the extract
(Bowman and Rand, 1980).
Blood pressure measurements reflect the status of the
cardiovascular system (Milnor, 1980), and the main-
tenance of an adequate blood pressure in the aorta
depends on the product of two factors, the cardiac output
and the total peripheral resistance of the vessels (Fried-
man, 1976). Therefore, the present study was focussed on
the effects of the crude extract from E. drupifera roots on
the blood pressure, heart and the aortic vessel. The
experimental results clearly point to the hypotensive
property of the extract, which is probably due to some
interference with the cholinergic transmission, although
Copyright # 1999 John Wiley & Sons, Ltd.
553
some sympathetic-like effect may not be completely
ruled out. This is suggested because blocking the
sympathetic innervation to the heart with propranolol (a
beta-adrenoceptor antagonist) failed to prevent extract-
induced depression of heart rate and blood pressure,
suggesting that the extract was acting at a different site.
Further evidence in support of this view is provided by
the observation that the crude extract significantly
depressed raised blood pressure and heart rate produced
by carotid occlusion. In addition, the extract was found to
prolong the effect of administered acetylcholine. Such
enhancement may also contribute to the observed
hypotensive activity of the extract. These findings
suggest that the roots of E. drupifera probably do not
interfere with the sympathetic transmission of the heart,
rather, enhancement of the cholinergic transmission is
strongly suspected as a mechanism. The finding that
atropine sulphate (a muscarinic cholinoceptor blocker)
antagonized the hypotensive and bradycardiac effects of
the extract is in line with this contention. Furthermore,
the extract induced a dose-dependent increase in the
contractions of the isolated guinea-pig ileum, which were
blocked by atropine sulphate (unpublished observation).
However, in the present experiments, whether atropine
antagonism was selective or not, was not studied, but
possibly, it was a competitive type since it was
surmounted by raising the concentration of the extract
(Fig. 1).
In the isolated tissue experiments, the ability of the
extract to relax aortic strips suspended in bath solution as
well as those precontracted with noradrenaline, suggests
that the extract has a direct relaxant effect on vascular
smooth muscle (Ajagbonna et al., 1995), and this may
further explain its blood pressure lowering mechanism.
However, the observation that the extract failed to relax
strips precontracted with KCl suggests that the effect of
the extract on vascular smooth muscle may be specific.
Such specificity could be directed to the alpha-
adrenoceptors. However, the action of phentolamine—a
non selective alpha adrenoceptor antagonist (McDevitt,
1979) in the presence of the extract (Fig. 5 d) is difficult
to explain since it failed to antagonize extract-induced
relaxation of the aortic strips. The result indicates a
possible extract–phentolamine interaction that could lead
to displacement of the potent agent(s) at the active sites.
This speculation, however, remains a subject for future
investigation.
In conclusion, it appears, therefore, that the crude
extract from roots of E. drupifera contains several
pharmacologically active constituents. One such group
may be acetylcholine-like agents showing predominantly
cholinergic activity, whereas the other constituents may
be catecholamine-like agents showing predominantly
alpha-adrenoceptor activity. However, given that the
active principle, let alone its chemical structure, is
unknown, further progress must await refinements in
the separation techniques.
Acknowledgements
We are grateful to Mr D.D. Dakat of the Physiology Department,
University of Jos, for technical assistance. Our thanks also go to Mrs
Nkoyo Nathaniel for typing the manuscript.
Phytother. Res. 13, 549–554 (1999)
554 A. E. ENO AND O. I. OWO
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