Research Journal of Applied Sciences, Engineering and Technology 2(3): 302-306, 2010
ISSN: 2040-7467
© M axwell Scientific Organization, 2010
Submitted Date: April 02, 2010
Accepted Date: April 13, 2010
Published Date: May 10, 2010
Body and Testicular Weight Changes in Adult Wistar Rats
Following Oral Administration of Artesunate
1
A.M . Izunya, 1 A.O . Nw aopara, 2 A. Aigbiremolen and 1 G.A. Oaikhena
1
Departm ent of Anatom y,
2
Departm ent of Pharmacology , College o f Medicine, A mbrose Alli U niversity,
Ekpom a, Edo State, Nig eria
Abstract: This experiment was designed to study the effects on the body and testicular weights of adult wistar
rats that recieved an oral administration of normal and double normal doses of artesunate. The rats were divided
into three groups (A, B and C) of five rats each. A and B served as the treatment groups, while C served as the
control group. Group A rats were given 4mg.kgG 1 b.w of artesunate daily for 3 days followed by 2mg.kgG 1 b.w
daily for next for 4 days. Group B rats were given 8mg.kgG 1 b.w of artesunate daily for 3 days followed by
4mg.kgG 1 b.w daily for next 4 days, while group C rats were given on ly distilled water. The rats were fed with
grow er's mash purchased from Edo feeds and Flour Mill Ltd, Ewu, Edo state and w ere given w ater ad libitum.
On day eight of the experiment, the rats were weighed and sacrificed . The testes w ere carefully dissected out,
freed from adherent tissues and weighed to the nearest 0.001 g. The results showed no changes in body weight
of rats in groups A, B and C. There was also no significant change in testicular weight of rats in group
A. However a significant increase in testicular weight was observed in group C. Our results suggest that
artesunate at norm al and double no rmal doses, has no effect on bo dy w eight of rats b ut may be toxic to the
testes at higher doses. It is uncertain however if these chan ges are reversible. It is recommended therefore, that
further studies aimed at corroborating these observ ations be carried ou t.
Key w ords: Antimalarial, artesun ate, body w eight, testicular w eight, tox icity
INTRODUCTION
Malaria is a leading cause of mortality and m orbidity
in developing areas of the world, and remains a major
public health problem in endemic regions (Breman et al,
2004. Resistance to available drugs is increasing, creating
a need for new drugs that are well tolerated and simple to
use. In the face of this ominous situation, artemisinin and
its derivatives (artesunate, artemether, arteether, and
dihydroartemisinin) have given renewed hope for
combating resistant malaria (Hein, 1994; Harinasuta and
Karbwang, 1994). These drugs have gained considerab le
prominence in the chemotherapy of both uncomplicated
and severe falciparum malaria by demonstrating high
activity against multidrug-resistant falciparum strains with
low toxicity profiles (Chanthap et al., 2005 ).
Artesuna te is one of the numerous drugs for malaria
intervention in Nigeria. It is a semi- synthetic derivative
of artemisinin, the active com pound of the Chinese herb
Artemisia annua which consists of the sodium succinyl
salt of dehydroartemisinin (Ittarat et al., 1999 ). Artesunate
and its active metabolite dihydroartemisin are potent
blood schizo nticidees, highly effective against multi-drug
resistant strains of plasmodium falciparum hence its
increasingly wide usage for the treatment and
management of malaria (Van Agtmaed et al., 1999 ). It is
used in com bination therapy an d is effec tive in cases of
uncomplicated P. falciparum.
Serious conc ern ha s been raised abou t unco ntrolled
use of artemisinin derivatives because at the moment they
are the last resort in the com bat against m ulti-drug
resistant P. falciparum malaria (White, 1994). The use of
these drugs should be controlled and restricted to proven
multi-drug resistance on severe ma laria in order to
preserve their efficacy (M uleng a, 199 8) and avoid
emergence of resistant strains. In malaria endemic areas
such as Nigeria, self medication is quite common and
purchase of antimalarials in the open market is rampant
(Nw anjo and Oze, 2007). The possibility of administering
overdose and misappropriation in the usage of
antimalarials are very common. D rugs though useful in
the treatment of disease conditions could also produce
untow ard effects in the individual. The untoward or toxic
effect m ay be harm ful to the patient (Udonan, 200 0).
Several studies have shown that high doses of
artesunate can produce neurotoxicity such as selective
Corresponding Author: Dr. A. M. Izunya, Department of Anatomy, College of Medicine, Ambrose Alli University, Ekpoma, Edo
State, Nigeria.
302
Res. J. Appl. Sci. Eng. Technol., 2(3): 302-306, 2010
damage to brainstem centres, gait disturbances in mice
and rats (Nontprasert et al., 1998, 2002; Genovese et al.,
2000) and loss of sp inal cord and pain response
mechanisms in animals (Genovese et al., 1995; Dayan,
1998). Others sho wed som e vary ing de gree o f cell
clustering, cellular hypertrophy, and intercellular
vacuolations in the stroma of the superior colliculus of
artesunate treated animals (Eweka and Adjene, 2008a).
Another showed some degenerative and necrotic changes,
cellular hyp ertroph y, and increa se interc ellular
vacuolations in the stroma of the stomach of rats ( Eweka
and Adjene, 2008b). More importantly, despite several
studies on the effects of artemisinin-based antimalarial on
male reproductive functions, there appears to be little or
no information in the literature on the impact of an ACT
drug, artesun ate, on body and testicular weights. This
study was therefore undertaken to examine the effects of
artesunate on the body and testicular weights of adult
wistar rats.
animals by orogastric tube for a period of seven days. The
dosage of artesunate was as per WHO recommendation of
4 mg.kgG 1 body weight daily for 3 days followed by 2
mg.kgG 1 body weight daily for the remaining 4 d ays. A ll
the animals were weighed before the experiment. The
drugs we re adm inistered to the groups as follows:
MATERIALS AND METHODS
Body and testicular weights determination: The
animals were weighed and autopsied by cervical
dislocation 24 h after the last the dose on the 8th day of
the respective trea tment. The testes we re dissected out,
freed from adherent tissues, and weighed up to the nearest
0.001g on a mettler analytical balance (PE 1600, Mettler
Instrum ent A G; Switze rland).
Group A: Four mg.kgG 1 body weight of artesu nate daily
for 3 days followed by 2mg.kgG 1 body weight daily for
the remaining 4 days.
Group B: Eight mg.kgG 1 body weight of artesunate daily
for 3 days followed by 4 mg.kgG 1 body weight daily for
the remaining 4 days.
Gro up C (Control): Distilled water.
The graded daily doses gave us the opportunity of
studying the effect of the normal and higher doses of the
drug.
Location and duration of study: This study was
conducted at the histology laboratory of the C ollege of
Medicine, Ambrose Alli University, Ekpoma, Edo State,
Nigeria. The preliminary studies, animal acclimatization,
drug procurement, actual animal experiment and
evaluation of results, lasted for a period of on e mo nth
(January, 2010). However, the actual administration of
the drug to the test animals lasted for one week (15,
January to 21, January 2010).
Statistical analysis: The data for body & testicular
weights were expressed as the m ean± SD . The treated
groups were compared to control using the Student’s t
test. Differences with values of p<0.05 were considered
statistically significant (M ahajan, 199 7).
Anim als: Fifteen adult wistar rats weighing between
100-150 g were used for this experiment. They were
obtained and maintained in the animal house of the
College of Medicine, Ambrose Alli University, Ekpoma,
Edo State. They were divided into three groups A, B, and
C of five rats each. Groups A and B were the treatment
groups, while G roup C served as the control. They were
kept in each group per cage and fed with grower’s mash
produced by Bendel Feeds and Flour Mills Limited, Ewu,
Nigeria. Water was given ad libitum. They were allowed
to acclimatize for one week before commencement of the
study. Ethical approval was sought and received from the
Department of Anatomy, Co llege of Medicine, Ambrose
Alli University, E kpoma, E do State on the need to
observe completely the rules guiding the employment of
rats for scientific studies.
RESULTS
General findings: No gross differences were observed
between the two groups of animals on day 8 at the
completion of experimental procedure. There were no
expected deaths recorded.
Effect of artesunate on body weigh t: The data obtained
from the mean body weights of the control and artesunatetreated rats are given in Table 1. A comparison of the
mean body weight of the rats before and after treatment
showed that both the artesunate-treated and contro l rats
manifested the same increase in body weight. The
percentage increase in mean bo dy w eight of rats
was 25% .
Drug administration: The artesunate tablets used for this
experiment, were manufactured by Mekophar Chemical
Pharmaceutical Join-Stock Company, Ho Chi Minh City,
Vietnam and purchased from Irrua Specialist Hospital,
Irrua, Edo State. The dru g solution w as made w ith
distilled water (1mg.mlG 1 ) and administered to the
Effect of artesunate on testicular w eight: The data
obtained from the mean testicular weights of the control
and artesunate- treated rats are given in Table 1. Using
t-test analysis technique there was no significant
difference in the mean testicular weights of rats in groups
303
Res. J. Appl. Sci. Eng. Technol., 2(3): 302-306, 2010
Tab le 1: Effect of arte sun ate on the body and testicular we ights of rats
following oral administration of normal and double normal
doses
Group
Body weight (g)
Testicular weight (g)
-------------------------------------------Initial
Fina l
% increase
A
100.00
125.00
25
0.606±0.365
B
100.00
125.00
25
*0.780±0.101
C
100.00
125.00
25
0.522±0.117
Values are expressed as mean± S D ; *: Sig nifica ntly d ifferen t statistica lly
from the c ontro l at p< 0.05 , T-tes t.
vascular abnormality (Bergh et al., 1988), inflammation,
salt retention and lymphatic obstruction (Robbins and
Cotran, 2004). It has also been shown that testicular fluids
reside in two compartments, the seminiferous tubules and
the interstitium (Setch ell et al., 1994). Seminiferous
tubule fluid is pro duced by the Sertoli cells, and its
volume is regulated by reab sorption in the efferent
ductules (Turner et al., 1984). Interstitial fluid originates
prima rily from the testicular vasculature, and its volume
is influenced b y cha nges in blood flow, vascular
permeability, physiological osm otic pressure differences,
and lymphatic drainage (Bergh et al., 1988). Testicular
interstitial fluid production has been shown to increase
after radiation (Laporte et al., 1985) and increase in
volume after the administration of drugs such as busulfan
(Udagawa et al., 2001), procarbazine (Delic et al., 1986)
and dibromochloropropane (D BC P) (Kluwe, et al., 1983).
Similarly it has been observed that blockage of the
efferent ducts by cells sloughed from the germinal
epithelium or the efferent ducts themselves can lead to an
increase in testis weight due to fluid accumulation (Hess
et al., 1991 ; Nak ai et al., 1993), an effect tha t could offset
the effect of depletion of the germina l epithelium on testis
weight.
Generally, artesunate exerts its anti-malarial activity
by generation of reactive, alkylating, oxygen free radicals
from its endoperoxide bond (M aggs et al., 1988) leading
to lipid peroxidation (Robert et al., 2001). The
accummulation of lipid peroxides is toxic to the
membrane structure , leading to a change in perme abilty
and to disintegration of cellular organelles (Muler and
Ohneso rge, 19 82). B ased on these reports therefore, it is
poss ible that the artesunate used in this study may have
acted in a similar manner to induce the testicular weight
gain observed in the double dose artesunate treated
group.
Moreover, the damage to the testes can be detected as
a weight change only at doses higher than those required
to produce significant effects in other measures of
gonadal status (Berndtson, 1977; Foote et al., 1986; Ku
et al., 1993). The effects seen in this study might have
been due to the ability of the artesunate to induce such
effects in the testes of the double d ose trea ted rats. .It is
uncertain however if these changes are reversible.
C and A, but there was a significance increase in mean
testicular weight of rats in grou ps C and B .
DISCUSSION
Our investigation on gross/morphological changes
indicated that there were no difference between the
control and experimental rats. The observed normal body
weight gain in the control and exp erimental groups,
implied that oral administration of norm al and double
normal doses of artesunate had no negative effect on
somatic growth. This finding agrees with the work of
Nw anjo and Oze (2007) in w hich a rtesun ate
administration caused no significant increase in body
weight of guinea pigs.
Although there was no difference in mean testicular
weight betw een the con trol and norm al dose artesu nate
treated rats, a significant increase in the mean testicular
we ight of the double normal dose treated rats was
however observed. In fact, it has been reported that an
increase or decrease in relative or absolute weight of an
organ after administering a chemical or drug is an
indication of the toxic effect of that chem ical (Simons
et al., 1995; Maina et al., 2008). Also, the weight of m ale
reproductive organs usually provides a u seful
reproductive risk assessm ent in experimen tal studies (Raji
et al., 2005; Simons et al., 1995) and testicular size is the
best primary assessment for spermatogenesis, since the
tubules and g ermin al elem ents ac count for approximately
98% of the testicular mass (Sherines and Ho wards, 1978).
Thus, the observed no-testicular-weight change in the
testis of both the control and the normal dose treated rats
sugg ests that the administration of artesunate at normal
dose had no toxic effect on this organ. But the observed
increase in testicular weight in the double normal dose
treated rats indicate that the drug may have toxic effect on
the testis at that dose.
Interestingly, there are studies that have shown that
the administration of a herbal tea mixture containing
Gynostemma pentaphyllum, Ra dix polygo ni multiflori,
Semen cassiae, Green tea and Folium nelumbinus caused
testicular oedema which has been linked to factors such as
testosterone (Maddock s and Sh arpe, 1989), Human
Chorionic Gonadotrophin (hCG) or endogenous LH
(Bergh et al., 1990), direct effect of GnRH agonists,
CONCLUSION
Our study suggests that artesunate at normal dose has
no effect on testicular weight but at double normal dose,
it causes a significant increase in testicular we ight. It is
however uncertain, if these chan ges are reversible. Thus,
there is a need to determine if these observations in wistar
rats may be applicable to humans and in this regard, one
can say that overdosage in humans may likely produce a
reproductive toxicological risk. W e therefore recommend
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Res. J. Appl. Sci. Eng. Technol., 2(3): 302-306, 2010
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