Pharm. Méd. Trad. Afr. 2004, Vol.13, pp. 175-187
Reversai of byperglycaemia and renal alterations in Streptozotocin diabetic
rats treated with Anacardium occidentale (Anacardiaceae).
Pierre Kamtehoulag", Leonard Tedong", Théophile Dlmn', Desire Paul
Ddjomeni Dzeuûet', Acha Emmanuel Assongalem2, Dogmo Selestin
Sokeng", Patrice Callard4 and Jean -François Flejou5•
1) Department of Animal Biology and Physiology, University of Yaounde l, PO
Box 812, Yaounde Cameroon.
2) Department ofPhysiological Sciences, Faculty of Medicine and Biomedical
sciences, University of Yaounde I, Cameroon.
3) Department ofBiological Sciences, Faculty of Sciences, University of
Ngaoundere, Cameroon.
4) Department ofPathology, Tenon Hospital, University of Pierre and Marie
Curie, France.
5) Department of Pathology, Saint Antoine Hospital, University of Pierre and
Marie Curie, France.
175

---

Pharm. Méd. Trad. Afr. 2004, Vol.l3, pp.175-187
Abstraet
Anocardium occidentale (Anaeardiaeeae) has recently ernerged as an effective oral treatment for diabètes.
Early morphological alterations in the rat kidney due to experimentally indueed diabetes by Streptozotocin are
described in order to evaluate the possible therapeutie role of hexane extract of Anacardium occidemale (AO).
Screening study in our laboratory had shown that hexane extract of AO is more potent in lowering blood glucose
levels in diabetie rats than aqueous. ethyl acetate, and methanol fraction. Our findings concem the alterations
observed in the rat kidney because this organ (together with retina) mainly involved in the early morphological
modifications associated with damage of organs and tissues. At the end of the treatment biochemical parameters
(total protein excretion rate, albuminuria, g1yeosuria, urea and glycemia) were measured. Photographs ofkidneys
were submitted to qualitative and quantitative analysis of images (using Quantimet 500 Image, Analyser-Leica)
equipped with specifie software). The following parameters were analysed: Diameter of Bowman's capsule.
distribution and total area occupied by glomerular capillaries, PAS positive structures (related in to the intensity
of reaction resulting from the different amount of muccopolysccharides ). Anacardium occidentale hexane
fraction (whieh contents, saponins, alkaloids, and polyphenols) is efficient in preventing these modifications
when treatment started earlier before diabetic damage of kidneys.
However, the histological study of mueopolysaceharides in diabetie animais highlighted, a tendeney for
kidneys to accumulate the substances. Anacardium signifieantly reduced the accumulation. Previous evidence
demonstrating a beneficiai effect of therapy based on hexane extract of Anacardium occidentale in insulin-
dependant diabetes mellitus seems to be confirmed by our experimental results in kidneys of adult rats. ln facto
Anacardium treatment is effective (in our experience) for ameliorating the morphological pattern observed early
in sorne diabetic tissues of rats. above ail. in the kidney.
Keywords: Anocardmm occidentale, Hexane extract, Diabetes mellitus, rats, Nephropathy.
1-1ntroduction
lnsulin-dependant diabetes mellitus (lDDM), also called type 1 diabetes is defined as a
chronic disease, characterised by a clinicat disorder of sugar, fat and proteins metabolism,
caused by absence of insulin to promote sufficient glucose output from the liver [1]. Patients
depend on insulin for survival. Without insulin, they develop acute metabolic complications
such as ketoacidosis and coma. Diabetic nephropathy is the most important cause of death in
type ] diabetic patients, of whom, 30010 - 40% eventually develop end-stage renal failure [2].
Studies have shown that good metabolic control is beneficial in slowing the progression of
nephropathy in diabetes, and if the duration of diabètes is prolonged before reinstitution of
nonnoglycemia, nephropathy is not easily reversed [3].
Experimental type 1 diabetes induced with streptozotocin in rats display many of the features
seen in human subjects with uncontro11ed diabetes me1litus [4]. The development of new
therapies that are able to improve glycaemia management and even to cure diabètes is of great
interest.
Anacardium occidemale L. (Anacardiaceae), plant originated from Brazil is used as folk
medicine in south of Cameroon and other African countries for the treatment of diabetes
me11itus, diarrhoea and hypertension [5].
176

---

Pharm. Méd. Trad. Afr. 2004, Va/.13, pp.175-187.
ln our laboratory, hypoglycaemic and protective role of this plant has been reported [6, 7].
Moreover, a preliminary test has revealed that hexane fraction is more potent in lowering
blood glucose than methanol, ethyl acetate and methylene choride.
Base on our previous findings, the current study aimed to identify biochemical and renal
histopathological abnormalities that occur with the evolution of streptozotocin-induced
diabetes in rats, and to appreciate their possible reversaI
after establishment of good
metabolic control with the hexane extract of Anacardium occidentale.
2-Materials and metbods
2-1 Plant
Plant materials of Anacardium occidentale were collected, in month of January and were
authenticated by comparison with national herbarium (Yaounde-Cameroon) file, voucher
specimen (N° 41935 IHNC).
The plant leaves were dried in the laboratory at room temperature and powdered in a mixer
grinder. The powder-dried leaves (3kg) were macerated in 8 J of methanol at room
temperature. After filtration, the solution was concentrated under reduced pressure. The
resulting extraet (262 g) was eluted in hexane (11) and concentrated to dryness to afford
viscous mass of hexane extract (35.67g) with the extraction yield of 13.6 %. 1.5 g of this
extract were dissolved in 3 ml of dimethyl sulfoxide (OMSO) and solution adjusted to 97 ml
with distiller water to obtain a solution of 100 ml.
2-2 Animals
Male wistar rats weighing 150-230g from the animal house of the faculty of science,
University of Yaounde 1, were used. The animaIs were fed with standard laboratory diet, and
given tap ad libitum. After randomisation into various groups, the rats were acc1imated for a
period of 6-7 days in new environment before the initiation of experiment. AnimaJs described
as fasted were deprived of food for at least 14 h, but had free access to water. A group of 5
non-diabetic (ND) rats receiving distiller water, was considered as control group whereas
another group of non-diabetic rats receiving extract of AO 300mg/kg was considered as
control treated group (NOT).
2-3 Streptozotocin-induced diabetic rats
Diabetes was induced in rats by single iv. injection of streptozotocin, Sigma chemical Co (St
Louis, Mo, USA) at a dose of 55 mg/kg (freshly dissolved in physiological saline), while non-
177

---

Pharm. Méd. Trad. Afr. 2004, Vol.13, pp.175-187
diabetic rats were injected with saline only. 72 h following streptozotocin injection, the blood
glucose was monitored, using glucometer Accutrend GC (Boeringer Mannheim, Germany).
Animais with plasma glucose levels higher than 200 mg/dl were selected for the study.
Oiabetic untreated rats were sub-divided into three groups: group 1 with two weeks duration
of diabetes (OC-2W), group II with 8 weeks duration of diabetes (DC-8W), and group III with
twelve weeks duration of diabetes (DC-12W).
ln the other hand, diabetic-treated rats were sub-divided into four groups: group IV, rats given
Aü 150 mglkg (O-2W+A0150), 2 weeks after diagnosis of diabetes; group V, rats given AO,
300 mg/kg (O-2W+Aü300), 2 weeks after diagnosis of diabetes; group VI, rats given AO,
300 mg/kg of BW immediately after diagnosis of diabetes (O-OW+Aü 300); group VU, rats
given Insulin, 2 weeks after diagnosis of diabetes (O-2W+ln).
Ali animais were housed individually, with free access to food and water. The plant extract
was given once daily for duration of7 weeks.
2-4 Determination ofbloodglucoselevel and other biochemical parameters.
To determine the blood glucose level, ail animais were ovemight fasted. Blood was obtained
from the tail vein and glycaemia was monitored once per week as previously described.
At the end of the treatment, ail rats were sacrificed , blood and organs were collected for
further analysis. Blood was centrifuged at 3000 XrPM at 4°C for ]0 min to separate the
serum. The level of glucose, albuminuria, glycosuria, and urea was measured using Randox
Kits.
2-5 Renal histological assessment
Nephropathy was assessed after treatment by measurements of 24-h urinary album in, total
protein excretion rate, glycosuria and urea. For urinary collection, rats were housed in
metabolic cages for 24 h. Seve rai drop oftoluene were added to the urine collection beaker to
inhibit microbial growth [8]. At the end of treatment, rats were sacrificed, kidneys were
removed, fixed in 10 % buffered formalin, dehydrated in gradua! ethanol (80-100°), cJeared in
xylene, and embedded in paraffin. Section (3 um thick) were prepared and then stained with
hematoxylin-eosine, Trichome of Masson, and PAS dye for photomicroscopic observation.
Photographs of kidneys were submitted morphometric analysis of images, using Quantimet
500 image, Analyser-Leica.
178

---

Pharm. Méd. Trad. Afr. 2004, Vol. 13, pp. 175-187
2.6 Statistical analysis
Ail values are expressed as means ± SEM. Statistical analysis were evaluated using student's t
test. Findings were considered to be statistically significant at a p value (*p) ofless than 0.05.
3-Results
The hexane extract exhibited antidiabetic property in streptozotocin-induced diabetic rats as
shown by glycemia and biochemical parameters levels measured in urine.
The effects of the extract on body and kidney weight in streptozotocin-induced diabetic rats
are given in table 1. The body weight significantly decreased (p<O.05) in diabetic rats
compared to normal controls at the end of the experiment. The kidneys weight were also
decreased in diabetic rats ( 19.54 %) as compared to healthy rats.
ln insulin as weil as extract (300 mg/kg given eartier or 2 weeks after diabetes diagnosis),
there were no significant decreased of the body and kidneys weight (Table 1).
The results of biochemical measurements used to evaluate renal function are shown in Fig 1.
By the end of study, total 24h urinary protein, albumin, glycosuria, and urea were
approximately, 7, 12, 100, and 4 times higher, respectively in diabetic controls animais
compared with non-diabetic rats. Regarding the effective dose (300
mg/kg)
given
immediately after diagnosis of diabetes, albuminuria, and urea were decreased with 24%, and
44%, as compared to diabetic untreated-rats. With the same comparison, we also observed a
decrease ofproteinuria and glycosuria with 9% and 26%, respectively.
The effects of the plant extract on serum glucose levels in streptozotocin diabetic rats are
shown in Fig 2. The initial blood glucose levels of the diabetic rats selected for the study were
in range of208 and 380 mg/dl. In the untreated control rats, the blood glucose level increased
by 328.8 mg/dl on the 14 days and 2111 diabetic rats died during this period. There after 4/9
animals died in the second period of experiment and the mean glucose levels on the last days
oftreatment in 5 animais, which survived was 400 mg/dl.
ln diabetic-treated rats, D-2W+AüI50 and D-2W+Aü300, the blood glucose levels steadily
decreased and it was 226.4 and 123.6 mg/dl, respectively at the end of the treatment. Thus, the
hexane extract of Aü restored (in the dose-dependant manner) the glycemia almost nearer the
normal values at the dose of 300 mg/kg. The effect of the plant extract (300 mg/kg) is there
for, comparable to that of insulin (standard drug). In the other hand, there were no significant
decreased of blood glucose levels in non-diabetic treated rats (NDT) with Aü 300 mg/kg
compared to non-diabetic rats receiving only distiller water.
179

---

Pharm. Méd. Trad. Afr. 2004, Vol. 13, pp.175-187
Phytochemical analysis revealed the presents of alkaloids, saponins and polyphenols ln
hexane extract of our plant.
The photographs in Fig 3B, C, and 0 show the histopathological lesions in glomeruli and
tubules. We observed an osmotic nephrosis, desquamation and destruction of tubular
epîthelium, pathologies, which increased with the duration of diabetes (Fig 3D). We also
remarked an atrophy of glomerular capillaries with Bowman's space dilated in morphometric
study (Table 1). Diabetic untreated-rats (D-8W), exhibited a significant atrophy of total area
occupied by glomerular capillaries, 3.5\\!m2 versus 6.38\\!m2 for non-diabetic rats. These
structures tended to be normal in Anacardium-treated animais. Periodic acid schiff (PAS)
staining revealed that mucopolysaccharide were abundant in diabetic rats Iddneys. Although
AO efficiently restored the histopathologicaJ and morphometric alterations (Fig 3E), it failed
to recover ail these failures when treatment started two weeks after diabetic state).
4-Discussion
The main funetion of the kidneys is to excrete the waste produets of metabolism, to regulate
the body concentration of water and salt in other to maintain the appropriate acid-base balance
of the plasma, and serves as an endocrine organ secreting erythropoietin, renin and
prostagladins [9].
ln this study, we have evaluated biochemical parameters determinants of progression to
nephropathy. Our observations confirm that microalbuminuria is a very good predictor of the
developing diabetic kidneys disease. However, the development of diabetic nephropathy is
characterised by a progressive increase in urinary prote in particularly albumin, and late
decline in glomerular filtration rate, leading eventually to end-stage renal failure [10].
Physiologically, the upper normal rang of urinary protein and albumin excretion is 150
mg/24h and 30 mg/24h, respectively [11-12]. Recently, Warram and al (13] studied a large
cohort of approximately 300 microalbuminuric type 1 diabetic subjects for 4 years and found
and increasing risk of developing nephropathy for patients with proteinuria.
Morphological study showed an atrophy of glomerular capillaries, interstitial fibrosis and
tubular necrosis, which increased with a duration of diabètes. Moreover, chemical analysis of
diabetic glomeruli indicates increased carbohydrate content in basement membrane. These
results are consistent with those of other authors analysing renal biopsies of human diabetic
patients, but with diffuse basement membrane thickening of capillaries, glomeruloscJerosis,
and exudative lesions even in diabetes with relatively short duration. [13].
180

---

Pharm. Méd. Trad. Afr. 2004, Vol. 13, pp.175-187
At the dose employed, we were able to demonstrate significant protection of hexane extract of
AO against early nephropathy (Proteinuria, albuminuria, and glycosuria). An interventions of
treatment slowed and sometimes inhibited the morphological alterations observed. However,
the excellent recovery of renal function expected with treatment of AO can he explained by
the regenerative capability of the renal tubules [14]. The action by which the extract lowered
the blood glucose is not wellknown. ft could act by increasing glycogenesis and/or decreasing
glycogenolysis in diabetic rat liver, there for, the mode of action of AO could he compared to
that of Ca,liSÙJ kleinii leaf in STZ-induced diabetic rats [15]. In another hand, the chemical
substances could be mediated by stimulation of regeneration process and revitalisation of
rema ining J3 celIs [16].
Phytochemical analysis had revealed the presents of alkaloids, polyphenols and saponins in
the plant extract. Based on increasing number of reports on blood glucose reduction
associated with sorne saponins [17] and alkaloids [18J, isolated from other medicinal plants it
is likely that the active principle (s) could be present in one or the two families chemical
substances. Accelerated chemical modification of proteins by glycoxidation and accumulation
of AGE (Advanced G1ycation End products) in tissue are implicated in pathogenesis of
diabetic nephropathy. The increase in AGE formation is sometimes attributed to increase
oxidative stress, which can he inhibited with polyphenols [19J.
Streptozotocin-induced diabetic rats are insulin deficient, hyperglycaernic, and have a
reduction in glucose uptake in adipose and skeletal muscle [20]. The effect of plant extract is
equal to (urea and albuminuria) or less than (proteinuria and glycosuria) that of insulin. But in
ail case, the hexane extract decreases the urinary parameters in the dose-dependant manner.
Insulin as a standard drug causes hypoglycaemia when taken in excessive doses and overt
hypoglycaemia is the most worrisome effect of this drug. Anacardium occidentale did not
cause hypoglycaemic effect and is not toxic, there tor, could serve for optimal protection of
renal function during diabetic nephropathy.
S-Conclusion
The goal ofthese studies was to evaluate the effect of variety period therapies on development
of renal complications in streptozotocin-induced diabetic rats. The experiments indicated that
the more effectively a therapeutic intervention limits the progression of nephropathy. Our data
shows that, the hexane fraction expressed the best protection against renal function during
nephropathy, and that alteration can be preventing by eartier treatment. Therefore, further
181

---

Pharm. Méd. Trad. Afr. 2004, Vol.l3, pp.175-187
studies are necessary to isolate the active component and to clarify in more detail the pathway
conceming the protection.
Ack~owledgements
This work was support by French Pathology-Cytology -Development (PCD) association.
Pr Lontsi David, Department of Chemistry, University of Yaounde 1, Cameroon, is gratefully
acknowledged, for having provided laboratory facilities tor the plant extraction.
6-References
l-Sanchez SS, Abregu AV, Aybar MJ, and Sanchez ANR. Changes in the expression of small
intestine extracellular matrix protein in streptozotocin induced diabetic rats. Cell. Biol.Inter
2000; 12: 1881-1888.
2-Giorgino F, Lavida L, Cavallo PP, Solnica B, Fuller J, and Chaturvedi N. Factors associated
with progression to macroalbuminuria in microalbuminuria type 1 patients: Eurodiab
prospective and complications study. Diabetologia 2004; 4: 1413-1418.
3-Renu A, Saiyada NA, and Odenbach S, EtTeet of reinstitution of good metabolic control on
oxidative stress in kidney of diabetic rats. .J Diabetes ('ompI2004; 5: 282-288.
4-Chattopadhyay S, Ramanathan M, Das J, and Bhattacharya SK. Animais models diabetes
mellitus.lndian.J Erp Biol 1997; 35: 1141-5.
5-Paris R, Plat M, Giono-Barber, Linhard J, and Laurens A. Recherche chimique et
pharmacologique sur les feuilles d'Anacardium occidentale L. (Anacardiacées). Bull Soc Med
Afr Noire 1977; 22: 275-281.
6-Sokeng SD, Kamtehouing P, Watcho P, Jatsa HB, Moundipa PF, and Lontsi D.
Hypoglycemie activity of Anacardium occidentale L. aqeous extract in normal and
streptozotocin-induced diabetic rats. Diabetes Res 2001; 36: 001-009.
7-Kamtchouing P, Sokeng DS, Moundipa PF, Pierre W, Jatsa BH, and Lontsi D. Protective
role of Anacardium occidentale extract against streptozotocin-induced diabetes in rats. Joum
ofEthnopharm 1998; 62: 95-99.
8-Alderson NL, Chachieh ME, Frizzell N, Canning P, Metz TO, Januszewski and al. EtTeet of
antioxidants and ACE inhibition on ehemieal modification of proteins and Progression of
Nephropathy in Streptozotocin diabetic rat. Diabetologia 2004; 47: 1385-1395.
182

---

Pharm. Méd. Trad. Afr. 2004, Vol. 13, pp.J75-J87
9-Bulloek BL. Pathophysiology: Adaptation and alteration function. 3n1 ed. Phyladelphya:
lB/Lippincott 1992; p. 623-630.
IO-Salah R, Pajiea P and Zijko M. Microalbuminuria and Diabetes Mellitus. l ïiabetlologia
Croat 2004; 33: 209-220.
l l-Morgensen CE. Microalbuminuria as predietor of clinical diabetic nephropathy. Kidney lnt
1987,3: 673-689.
12-Warram JH, Scott U and Hanna LS. Progression of microalbuminuria in type 1 diabetes:
non linear relationship with hyperglycemia. Diabetes 2000 49: 94-100.
13-Kissane JM. Anderson's pathology, 81h edition. Toronto: Wshington University Sehoo1 of
Medicine; 1985. p. 754-9.
14-Babu V, Gangadevi T and Subramoniam A. Antidiabetic activity of ethanol extract of
cassia kleinii leaf in streptozotocin-induced diabetic rats and isolation of active fraction and
toxicity evaluation of the extract. lndianJ of pharmacol 2003; 35: 290-296.
]5-Diatewa
M,
Samba
BC,
Assah
HCT,
and
Abena
AA.
Hypoglycemie
and
antihyperglyeemic effects of diethyl ether fraction iso1ated from the aquous extract of the
leaves of cogniauxia podoleana Baillon in normal and alloxan-induced diabetic rats. .1 of
h'tnopharmaco/2004; 92: 229-232.
16-Bolkent S, Yamardag R, Tabakoglu-oguz, and Ozsoy-sacan, O. Effects of chard (beta
vulgatis L. var. cicla ) extract on panereatic 13 eells in streptozotocin-diabetic rats: a
morphologie and biochemical study. .l ofEthnopharmacol Iïïôî); 73: 251-259.
17- Marles RJ and Famsworth NR. Plants as sources of Antidiabetic Agents. Econom and
Med Plant Res 1994; 6: 150-185.
18-Raj DSC, Choudhury D, Welboume TC, Levi M. Advanced glyeation end produets: a
nephrologist's perspective. Am.J Kid Dis 2000; 35: 365-380.
19-Hennebele T, Salpaz S, Baileuil F. Polyphenols vegetaux, sources utilisations et potentiel
dans la lutte contre le stress. Phytotherapie 2004; 2: 3-6.
20-Chavabarti Rand Rajagopalan R. Diabetes and insulin resistance associated disorders:
diseas and therapy. Curr Sc 2002; 83: 1533-8.
-Cresponding author:
Leonard Tedong
Animal Physiology Laboratory
Faculty of Sciences
University of Yaounde 1
PO Box 812 Yaoundé- Cameroon
Itedot1g@y~hoo.fr
Itedong@uycdc.uninet.cm
183

---

r-,
00
........
1
~
........
~
"'":Î
........
-.;
~
~
~
'""t
'<:t
00
.....
ND
ND-A0300
D-8W
~
D2W+A0150
D2W+A0300
O-OW+A0300
D-2W+ln
"'l::i
Relative percentage of
0.38
0.37
038
0.44
0.37
0.38
~
0.35
~
the kidneys weight
UER m1l24h
3.6±1.02
8.6±O.S2
62.8±4,7*
4S.8±8,4*
30.8±0.8*
18.75±1.1
20.00±1.1
"'l::i
'~
Total surface of
8.71±O,3
8.5±1.04
S.78±1.2*
~
6.6±O.38
7.6±1.06
70S±D.14
7.17±O.49
~
glomeruli n=5 (urn")
Total area occupied
6.38±0,13
6.44±1.02
3.SO±O.8
~
S.10±O.43
4.37±O.73
4.8S±0.31
S.22±0.39
~
by glomerular
capillaries 0=5 (ll_m_2.!-)
_
Table 1 Physical parameters in fasted rats, 35 days after treatment with hexane
extract of Anacardium occidentale. Diabetic control rats (D-8W) showed a significant decrease in total area occupied by glomerular
capillaries, compared with non dîabetic rats (ND). Values shown are mean ± SEM, *P< 0.05, significant different comparee te control
rats. UER: Urinary Excretion Rate. n: number of glomeruli measured.

---

Pharm. Méd. Trad. Afr. 2004, Vol. 13, pp.175-187
~OND
-NDT
mO-8W
1
I O NO
• NOT
oD-8W
l'
l /1IijJ D-2W+A0 150 1lI0-2W+A0300 Oo-OW+A0300
!œD-2W+A0 150 1iilD-2W+A0300 OD-OW+A0300! 1
. 1
,
lIo-2W+~__
~
II!! D-2W+/n
I
i 1,
. ~ - - _ .
1
.
1
1
*a
1
1
Ra
100
1
8
*b
~ 90
l' .!
7
l!
~
80
1
c
6
:E
70
*e
Cl
:8 - 5
g 60
e~
1
U N
4
50
.!!
1(::0
3
Glèl
5
40
1
c -
c
30
'Ë
I ~ 2
:::l
20
Q.
1
10
o
0""--'==
A-Experimentaf groups
B-Experlmental groups
L-- - - -'==ïl
OND
.M1T
BD-8W
1 1
mND
.NDT
E1D-8W:~
D D-2W+A0150 /1 D-2W+A0300 0 D-OW+A0300
1ID-2W+ln
rI!IIID-2VV+A0150iiiD-2VV+A030013[).M+A0300 1
(gD-2VV+/n
1
-
-
,
1
1
140
*a
1
*b
120j
200
180
100 1
_160
J;;
140
80
*e
*e
~ 120
60
E 100
-;;
80
40
~
60
;:)
40
20 '
20
O-""'-'-========== =---
o
c -experimentai groups
D-&perimental groups
l_ .
_
Fig. 1 Metabolic parameters in urine collection of fasted rats, 35 days after treatment with Anacardium
occidentale. A : Proteinuria ; B : Albuminuria ; C: Glucose excretion rate; 0 : Urea. Oiabetlc control
rats (D-8W) showed a marked increase in ail melabolic parameters compared with non-diabetic rats (
ND). Values shown are the mean ±SEM, *P<0,05, Signiticant difference as compared to, control (a),
Anacardium-treated group (b) and Insu/in (c).
185

---

Pharm. Méd. Trad. Afr. 2004, VoU 3, pp.l75-l87
-+-ND
j
j
-NOT
-A-O-aW
:;
-D-2W+AD 150
-+- 0-2W + AD 300
-+- O·OW + AD 300
1)
-D-2W+ ln
!
:__ .-._._--_.-~---_.--_._~-- -~--~ - - _.. _---_._-.---~-_._--_._-------
500 ,
*
=- 450 ,
1
32 400 J
= :
E
1
- 350 ~
~ 300 -:
..!!
1
Q)
250 ..,
li)
0
!
CJ
200 .;
.a
1
= 1
150
"a
-t
#,
0
0
100 l
iD
50 -i
!
1
- - - - - - - - - - - . ~ ~ - - - . - - - -
••- - - .
1
o 'r--
1
OW
W1
W2
W3
W4
W5
1
Time (weeks)
Fig 2 Effects ofhexane extract ofAnacardium occidentale on blood glucose
levels of streptozotocin-diabetic rats. Data are shown as means ± SEM. *p<O.OS,
significant different compared to non diabetic (ND) rats, and #, significantly
different frOID the initial day oftreatment, p<O.OS.
186

---

Pharm. Méd. Trad. Afr . 2004, VoU3, pp. 175-187
13
Fig.3: PAS stain of kidneys of rats at 35 days and 28 days of treatment age with hexane
extract of Anacardium occidentale and Insulin, respectively.
A: Non diabetic (ND) rat. B: Diabetic control rat on 2 weeks (DC-2W) after diagnosis of
diabetes. C: Diabetic control rat on 8 weeks (D-8W) . 0 : Diabetic control rat on 3 months
(DC-3M). E: Diabetic rat on zero day, given AO 300mg/kg/day. F:Diabetic rat on 2
weeks,given AO 300mg/kg (D-2W+AO 300).G: Diabetic rat on 2 weeks, given insulin,
5lJUlkg/day (D-2W+ln) ( - + ) arrows indicate epithelium tubules nephrosis. (~
)arrow indicates positive structures from the different amount of muccopolysaccharides
187

---