Acute Toxicity and Antipyretic Activities of a Methanolic Extract of <i>Alchornea cordifolia</i> Leaves

doi:10.4236/pp.2013.47A2001

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Pharmacology & Pharmacy, 2013, 4, 1-6

http://dx.doi.org/10.4236/pp.2013.47A2001 Published Online October 2013 (http://www.scirp.org/journal/pp) 1

Acute Toxicity and Antipyretic Activities of a Methanolic

Extract of Alchornea cordifolia Leaves

K. E. Effo1*, G. Kouakou-Siransy1, G. Irie-Nguessan1, R. W. Sawadogo2, I. L. Dally3, A. B. Kamenan1,

L. S. Kouakou1, J. Kablan-Brou1

1 Laboratoire de Pharmacologie et de Physiologie, UFR des Sciences Pharmaceutiques et Biologiques, Université Félix Houphouët

Boigny, Abidjan, Côte d’Ivoire; 2Laboratoire de Pharmacologie et de Toxicologie, Institut de Recherche en Sciences de la Santé

(IRSS/CNRST), Ouagadougou, Burkina Faso; 3Laboratoire de Galénique, UFR des Sciences Pharmaceutiques et Biologiques, Uni-

versité Félix Houphouët Boigny, Abidjan, Côte d’Ivoire.

Email: *[email protected]

Received August 9th, 2013; revised September 12th, 2013; accepted September 28th, 2013

permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

ABSTRACT

Alchornea cordifolia (Euphorbiaceae) is a very prized plant among traditional healers in Africa. Its leaves are used for

its antipyretic properties in traditional areas. The aim of our study is to determine the acute toxicity and the antipyretic

activity of a methanolic extract of Alchornea cordifolia leaves. Acute toxicity was assessed by measuring mortality,

changes in body weight, spontaneous movements, and normal rectal temperature in mice. Antipyretic activity was

evaluated by brewer’s yeast-induced hyperpyrexia in rats according to Teotino method (1963). The antipyretic effect of

methanolicextract of Alchornea cordifolia leaves was compared with paracetamol (100 mg/kg bw) orally. Groups of

mice treated with doses of 6500; 3250; 1625 and 812.5mg/kg of the extr act did not show any mortality, nor sign ificant

alteration of body weight, nor alteration of spontaneous movements. However, incomplete reversed dose-dependent

hypothermic activity was observed with doses of 50.78; 101.56; 203.12; 406.25; and 812.5 mg/kg p.o. of the extract,

showing acute toxicity of this plant. In the antipyretic assay, the extract with doses of 50.78; 101.56; 203.12; 406.25;

and 812.5 mg/kg p.o. exhibited a significant dose-dependent antipyretic activity similar to paracetamol (100 mg/kg bw)

in rats. Thus Alchornea cordifolia may inhibit prostaglandins-biosynthesis from hypothalamus. Our results support

claims on its traditional uses in management of fever. However Alchornea cordifolia may aff ect hypothalamus not only

during fever but also when body temperature is normal.

Keywords: Alchornea cordifolia; Acute Toxicity; Brewer’s Yeast; Hypothermia

1. Introduction

Alchornea cordifolia (Euphorbiaceae) is a plant that is

widely used in traditional medicine in Africa, from Sene-

gal to Cameroon, as a remedy for several diseases. Its

leaves are renowned for being febrifuge [1]. Previous

studies revealed no mortality in mice in acute toxicity

tests, several pharmacological activities such as antim-

icrobial [2-4], antiameobic [5], antimalarial [6], anti-in-

flammatory [7,8], antispasmodic [9], antioxidant [10],

antidiarrheic [11], and anti-stress properties [12]. How-

ever, there is no existing pharmacological data that indi-

cated the antipyretic activity o f Alchornea cordifolia. T h u s,

the aim of our study was to determine the acute toxicity

and the antipyretic activity of the methanolic extract of

Alchornea cordifolia leaves in rats.

2. Materials and Methods

2.1. Plant Material

Fresh Alchornea cordifolia (Schum. and Thonn.) leaves

were harvested in the Floristic National Center of Abid-

jan and the collected leaves were given to the botanic

expert Prof Aké-Assi Laurent affiliated with the Univer-

sity of Cocody (Abidjan), who verified their uniformity

with the specimens previously deposited and catalogued

in the herbarium of the Floristic National Center of Abid-

jan. The leaves were collected and dried in darkness in a

25˚C air-conditioned room

2.2. Experimental Animals

Mice (Mus musculus) (15 - 34 g) and rats (Rattus

norvegicus) (weighing 150 - 286 g) were purchased from

*Corresponding author.

Acute Toxicity and Antipyretic Activities of a Methanolic Extract of Alchornea cordifolia Leaves

Department of Nutrition and Pharmacology of the Uni-

versity of Cocody (Abidjan, Côte d’Ivoire). The animals

were then bred at the Department of Pharmacology of the

faculty of pharmacy of the University of Cocody. All

animals were kept in the room maintained under envi-

ronmentally controlled conditions of 24˚C ± 1˚C and 12

h light - 12 h dark cycle. The animals had free access to

water and food and were acclimatized at least 1 week

before starting the experiments. The animals were cared

for and treated according to the principles of care for, and

use of, laboratory animals, formally approved by the Bo ar d

of Ethics of the Cocody University, in compliance with

the Guide for Care and Use of Laboratory. And before

the experiments they were fasted overnight with water ad

libitum.

2.3. Chemicals

Metha nol (Si g m a ), so di um car b oxy -m e thyl -c el lul o se (CMC)

Sigma (France). Paracetamol: (Doliprane 100 mgpoudre-

pour solution buvable) (Sanofi Aventis), brewer’s yeast

Arkogélules (Arkopharma) were used in this study.

2.4. Methanolic Extract

The fine powder (200 g) of dried leaves were macerated

for 24 h at room temperature in 700 ml of methanol. The

methanolic extract obtained were dried in a rotavapor

(Büchi R180) and conserved at 4˚C, and aliquo ts of dried

powder were used for pharmacological studies after sus-

pending in 2% CM C.

For acute toxicity test, the stock solution was concen-

trated at 650 mg/ml. Three other ranges of concentration

were prepared from this stock solution by successive

dilutions in 1/2 (325 mg/ml, 162.5 mg/ml, and 81.25

mg/ml).

We used successively for th e hypothermic activity and

antipyretic activity, solutions concentrated at 81.25 mg/ml,

40.62 mg/ml, 20.31 mg/ml, 10.15 mg/ml and 5.07 mg/ml

2.5. Acute Toxicity Test

Mortality, body weight changes and spontaneous motor

activity were assessed in mice and hypothermic activity

in rats.

2.6. Spontaneous Motor Activity (SMA)

A multi-counter activity cage, Letica activity cages (LE

886) connected to AM1051 (Benwick electronics) data

logger were used for this study. The AM1051 data logger

is provided with two layers of infrared sensor placed

horizontally to monitor the rearing, mobile and static

activities, as well as the active and mobile times. Mice

were singly placed in each cage and activity was auto-

matically recorded by the instrument for 5 min. Each

mouse was used only once. The extracts (50.78; 101.56;

203.12; 406.25; and 812.5 mg/kg p.o.) were adminis-

trated to five groups of mice (n = 6) and control group

received CMC. SMA measurements started one hour after

the administration of the extract and the fourth hour and

subsequently every two days for 14 day s after treatment.

2.7. Effect on Normal Body Temperature

(Hypothermic Activity)

Rats weighing 150 - 286 g were divided into six groups

of six animals. Initial rectal temperatures were recorded

using a 12 channel electric thermometer (LETICA Scien-

tific instruments), model 812 RS, PANLAB s. L, Barce-

lona. Then extract (50.78 mg/kg; 101.56 mg/kg; 203.12

mg/kg; 406.25 mg/kg; 812.5 mg/kg) was given orally.

Temperature changes in body temperature values before

and after drug administration were recorded.

2.8. Brewer’s Yeast-Induced Hyperthermia in

Rats

Rats weighing 180 - 220 g were divided into six groups

of six animals. Initial rectal temperatures were recorded

using a 12 channel electric thermometer (LETICA scien-

tific instrument), model 812 RS, PANLAB s. L, Barce-

lona. Hyperthermia was induced in rats according to the

method of Teotino et al. [8] by subcutaneous injection of

1 ml/100g b ody weigh t of 20% brewer’s yeast. When the

temperature was at a peak, 18 h after yeast injection, only

rats which developed satisfactory pyrexia (0.6˚C or more

increase in rectal temperature) were used. Then extract

(50.78 mg/kg; 101.56 mg/kg; 203.12 mg/kg; 406.25 mg/kg;

812.5 mg/kg) was given orally 24 h after yeast injection.

Paracetamol (100 mg/kg, p.o.) served as the reference

drug for comparing the antipyretic action of extract and

the rectal temperatures of animals were recorded at 1h

interval for 4 h following drug treatment.

The results are expressed as percentage [13,14]:



Percent reduction

y east-induced pyrexiapost-treatment tem perature10 0

Yeast-induced py rexia





2.9. Statistical Analysis

The results are expressed as mean value ± S.E.M. Fried-

man’s test was applied to the results. Mean values were

considered significantly different when P < 0.05.

3. Results

3.1. Acute Toxicity

Mortality and body changes weight

No death was observed nor a significant changes in

Acute Toxicity and Antipyretic Activities of a Methanolic Extract of Alchornea cordifolia Leaves

weight (Table 1). administered at 812.5 mg/kg to the rat presented an anti-

pyretic effect superposable to that of the paracetamol

administered at 100 mg/kg. The doses of 203.125 mg/kg,

406.25 mg/kg presented also a statistically significant

antipyretic activity (p < 5%).The antipyretic effect is

totally reversed after 4 hours.

Spontaneous motor activity

After a two-week observation, the methanolic extract

from Alchornea cordifolia engendered no sign ificant mo di -

fication of spontaneous motor activity.

Effect on normal body temperature

The doses of 203.125 mg/kg, 406.25 mg/kg and 812.5

mg/kg, presented hypothermic activity (p < 5%). The

methanolic extract of Alchornea cordifolia causes a hy-

pothermia that is not totally reversed at the end of a

four-hour time (Figure 1).

Figure 3 shows the percentage of hyperthermia inhibi-

tion after two hours. We noted a reduction of the hyper-

thermia upper to 80% with 203.125 mg/kg, 406.25 mg/kg

and 812.5 mg/kg. In doses lower than 203.125 mg/kg, we

noted a percentage of reduction lower than 50%.

4. Discussion

3.2. Antipyretic Activity

Figure 2 show the antipyretic activity. During four (04)

hours, the methanolic extract of Alchornea cordifolia Alchornea cordifolia is a plant which is very currently

used in traditional medicine for the treatment of several

Table 1. Mortality and body changes weight.

Substance mice number at the beginning mice number at the end Average initial weight (g) Average weight at the end (g)

CMC 2% 8 8 22.4 24.5

MEAC: 81.25 mg/kg 8 8 22.2 22.4

MEAC: 162.5 mg/kg 8 8 22.2 23.7

MEAC: 325 mg/kg 8 8 22.3 22.3

MEAC: 6500 mg/kg 8 8 22.2 23.6

Values rep resent the average ev olution of the number of mice an d the average o f weight. Number o f animals used ( n = 8). *p < 0.005 compared to control by

Friedman’s te s t. *: P < 0.05: significantly diffe r ent compared t o control (CMC 2%).

33.5

34.5

35.5

36.5

37.5

T0h T1h T2h T3h T4h

Rectal temperature (°C)

Time interval

CMC 2%MEAC: 50.78 mg/kgMEAC: 101.56 mg/kg

MEAC: 203.12 mg/kgMEAC: 406.25 mg/kgMEAC: 812.5 mg/kg

***

MEAC: Methanolic extract of Alchornea cordifolia; CMC: carboxyméthylcellulose Curves represent the

average evo lutio n of the r ectal temper atur e in ti me. Number o f animals used (n = 6 ). *p < 0.005 compared

to control by Friedman’s test. *: P < 0.05: significantly different compared to control (CMC 2%).

Figure 1. Hypothermic activity of methanolicextract of Alchornea cordifolia.

Acute Toxicity and Antipyretic Activities of a Methanolic Extract of Alchornea cordifolia Leaves

33.5

34.5

35.5

36.5

37.5

T-16hT0h T1hT2h T3hT4h

Rectal temperature (°C)

Time interval

CMC 2%MEAC: 50.78 mg/kgMEAC: 101.56 mg/kg

MEAC: 203.12 mg/kgMEAC: 406.25 mg/kgMEAC: 812.5 mg/kg

Paracétamol: 100mg/kg

Curves r epresent the av erage evo lution o f the rect al temperat ure in t ime. Number o f animals u sed (n = 6).

*p < 0.005 compared to control by Friedman’s test. *: P < 0.05: significantly different compared to control

(CMC 2%).

Figure 2. Antipyretic activity of methanolicextract of Alchornea cordifolia and paracetamol on brewer’s yeast induced

pyrexia in rats.

100

120

140

160

180

Inhibition of hyperthermia (%)

Treatment

The result s are given are mean ± S .E.M. number of animals used (n = 6). *p < 0.005 compared to control

by Friedman’s test.

Figure 3. Inhibition of hyperthermia ofmethanolicextract of Alchornea cordifoliaand paracetamol (100 mg/kg) at two hours.

diseases associated with fever. Our works aimed to s ear ch

out a possible toxicity and an antipyretic activity. About

acute toxicity, absence of death in the present study was

also observed by Traoré [15] using 5000 mg/kg p.o. of

leaves orally and by Umukoro [12] using 4000 mg/kg

orally. The absence of modification in the spontaneous

movements shows that Alchornea cordifolia cannot mod-

ify the normal behavior in mice. However, the dose-de-

pendent hypothermia observed in the present study after

administration of Alchornea cordifolia extract showed

acute toxicity of this plant. Hypothermia indicated altera-

tion of preoptic anterior hypothalamus which is critical in

Acute Toxicity and Antipyretic Activities of a Methanolic Extract of Alchornea cordifolia Leaves 5

the neuronal network of thermoregulation. The hypothala-

mus regulated the set point at which body temperature is

maintained. Hyperthermia inhibition observed in the pre-

sent work varies from 89% to 160%. Evolution of hyper-

thermia inhibition due to Alchornea cordifolia at 812.5

mg/kg during four hour was similar to paracetamol (100

mg/kg p.o.) with a pick at two hours (Figure 2). The

mechanism of the induced hyperthermia by brewer’s

yeast is mediated by prostaglandins biosynthesis [16,17].

Brewer’s yeast administrated through subcutaneous route,

behaves lik e an exogenous pyrogen. Thu s, it produces an

inflammatory process that according to Milton et al. [18]

and Aronoff et al. [19], leads to the biosynthesis of

chemical mediators as prostaglandins due to arachidonic

acid. Among prostaglandins, PGE2 is strongly involved

in the disturbance of the hypothalamic thermostat and so

is responsable for fever [18,19]. Thus, the antipyretic

activity of Alchornea cordifolia observed in this study

would be related to the inhibition of the production of

prostaglandins. Apart from their involvement in fever,

prostaglandins are also known for their vasodilatation

effect that generates red blotch and oedema as inflamma-

tion characteristics. Thus the hypothesis about the inhibi-

tion of the production of prostaglandins by Alchornea

cordifolia could be uphold with the works by Osabede

and Okoye [8] as well as those by Mavar-Manga et al. [7]

highlighting the protecting effect of Alchornea cordifolia

against oedema, the inflamemation model induced in

rodent.

These antipyretic property lay in the saponins contain

in Alchornea cordifolia leaves [5,9] that, according to

Gepdiremen et al. [20] are strong prostaglandin inhibi-

tors.

5. Conclusion

This antipyretic activity that is similar to paracetamol

one could partly justify the traditional use of Alchornea

cordifolia leaves against fever in the African societies.

Considering the partially reversible hypothermic effect in

a 4-hour observation time, we plan a more elaborate

study on the neurotoxicity of that plant.

6. Acknowledgments

The authors are grateful to Pr. L. Aké-Assi (Centre Na-

tional de Floristique d’Abidjan) for botanical identifica-

tion.

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