ArticlePDF Available

SOME EFFECTS ASSOCIATED WITH THE USE OF THE BIOPREPARATION FROM Picralima nitida SEEDS EXTRACT AS ANTIDIABETIC AGENT

Authors:
  • Federal University of Agriculture, Abeokuta

Abstract and Figures

The study was aimed to investigate some untoward effects that could be associated with the use of P. nitida as hypoglycemic agent using some biochemical and histological evidences. The antioxidant property of the plant was determined by using 1, 1-diphenyl-2-picrylhydrazyl radical scavenging activity. Biochemical studies in plasma using determining the testes such as blood glucose, alanine and aspartate aminotransferases, gamma glutamyl transferase activities, electrolytes (sodium, potassium and bicarbonate, lipid peroxidation levels, haematological parameters (red blood cell and whole blood cell, platelets, and lymphocyte counts), blood glucose level, lipid profile, and also liver and kidney function tests were performed. Histopathological examinations of the liver, kidney and pancreas were done following the standard Heamatoxylin and Eosin staining method. Methanol extract of the seeds has the highest antioxidant level (36.73%), indicating higher free radical scavenging activity; followed by aqueous extract (19.36%) and coconut water extract (4.09%). There was significant reduction (p
Content may be subject to copyright.
BIOTECHNOLOGIA ACTA, V. 7, No 2, 2014
92
Diabetes is becoming one of the prevalent
disease conditions worldwide. Diabetes melli-
tus, often simply referred to as diabetes, is a
group of metabolic disease in which a person
has high blood sugar, either because the body
does not produce enough insulin, or as a result
of insensitivity of target cells to the insulin
that is produced [1]. Clinically, there are three
main types of diabetes: Type 1 diabetes melli-
tus, also known as Insulin Dependent Diabetes
Mellitus (IDDM) or juvenile onset diabetes is
caused by an autoimmune destruction of the
insulin secreting β-cells of the pancreas [2]. It
usually develops during childhood, adolescence
or during early adulthood [3], since the insulin
producing β cells are partially or completely
loss, the patient requires daily injection of in-
sulin [4]. It is also characterized by a condition
in which the pancreas does not produce insulin
at all, which could be as a result of damage on
UDK 616.379-008.64 doi 10.15407/biotech7.02.092
SOME EFFECTS ASSOCIATED
WITH THE USE OF THE BIOPREPARATION
FROM Picralima nitida SEEDS EXTRACT
AS ANTIDIABETIC AGENT
Key words: Picralima nitida, diabetes mellitus, biochemical, histological evidences.
O. A. Akinloye1 1Department of Biochemistry, Federal University of Agriculture,
E. A. Balogun2 Abeokuta, Nigeria
S. O. Omotainse1 2Department of Veterinary Pathology, Federal University of Agriculture,
O. O. Adeleye1 Abeokuta, Nigeria
Email: oaakin@yahoo.com
Received 23.10.2013
The study was aimed to investigate some untoward effects that could be associated with the use of
P. nitida as hypoglycemic agent using some biochemical and histological evidences.
The antioxidant property of the plant was determined by using 1, 1-diphenyl-2-picrylhydrazyl radical
scavenging activity. Biochemical studies in plasma using determining the testes such as blood glucose,
alanine and aspartate aminotransferases, gamma glutamyl transferase activities, electrolytes (sodium,
potassium and bicarbonate, lipid peroxidation levels, haematological parameters (red blood cell and
whole blood cell, platelets, and lymphocyte counts), blood glucose level, lipid profile, and also liver and
kidney function tests were performed. Histopathological examinations of the liver, kidney and pancreas
were done following the standard Heamatoxylin and Eosin staining method.
Methanol extract of the seeds has the highest antioxidant level (36.73%), indicating higher free
radical scavenging activity; followed by aqueous extract (19.36%) and coconut water extract (4.09%).
There was significant reduction (P < 0.05) in blood glucose of all the treated rats at the end of the exper-
iment (ranging from 41.66% to 55.59%). Significant increase (P < 0.05) in body weights of the treated
rats were also observed at the end of the treatment (ranging from 9.26% to 38.89%). There was a sig-
nificant (P < 0.05) increase in the hematological parameters in all the extract treated groups. There was
also significant decrease (P < 0.05) in the lipid profiles of the treated groups. Plasma studied enzymes
activities decreased in all treated groups. Ionoregulatory disturbances observed included hyperkalemia
and hypernatremia in all the treated groups but were reduced significantly (P < 0.05) at the end of the
treatment. Urea and bicarbonate concentrations and also of lipid peroxidation level decreased signifi-
cantly in all the groups. The histopathological studies revealed that the extracts were unable to amelio-
rate some observable pathologic conditions associated with induced diabetic tissues. Although, diabetes
mellitus have been reported to be associated with varied histological changes in different organs, in this
study, histological examinations of the pancreas of the treated and untreated groups showed varying
degree of degenerations but the extent of severity in the lesions were more pronounced in the extract
treated groups. In this relation the obtained results of this study which revealed the hypoglycemic and
antioxidant potentials of Picralima nitida seed extracts for the treatment of diabetes mellitus should be
taken with caution in administering the P. nitida seed extract as an hypoglycemic agent.
Experimental articles
93
the pancreatic cells. Type 2 is often referred to
as Insulin Independent Diabetic which is char-
acterized by a condition in which the pancreas
does not produce enough insulin or the insulin
produced is not working properly. Gestational
diabetes is when pregnant women who have
never had diabetes before have high blood glu-
cose level during pregnancy, in some cases it
may precede development of diabetes type 2
[2]. Long term diabetes mellitus is commonly ac-
companied by other cardiovascular risk factors
such as dyslipidemia, hypertension, prothrom-
bic factors and microvascular problems involv-
ing eyes, kidney and peripheral nerves [5].
Treatment of diabetes is very important
because it is a life threatening ailment and
presently there is no known cure [6]. Manage-
ment of diabetes often concentrates on keeping
blood sugar levels close to normal (euglycemia)
as possible without presenting undue danger
to the patient. Dietary management, regular
exercise, and use of appropriate medications
are some other management regime. Long
term diabetes mellitus is commonly accompa-
nied by other cardiovascular risk factors such
as dyslipidemia, hypertension, prothrombic
factors, ketoacidosis, and non-ketotic hyper-
sosmolar coma and microvascular problems
involving eyes, kidney and peripheral nerves
failure, poor wound healing, gangrene on the
feet which may lead to amputation and erectile
dysfunction or impotence [5, 7].
Picralima nitida is a member of the fami-
ly Apocynaceae and order gentianales and is
widely distributed in the tropical rainforests
of Africa, as homesteads or bushes [8, 9]. It
can measure up to 20–35 m high when fully
grown, having white flower in terminal inflo-
rescence and very large paired fruits (pods).
Its leaves are opposite, simple, and entire.
The fruit (pod) has many seeds which are usu-
ally embedded in the pulpy material known as
pulp. The pericarp of the fruit which contains
latex is known as the rind [10]. P. nitida seed
is known as Akuamma seed in Ghana, Osi-igwe
seed in Igboland (Eastern Nigeria), Eso Abere
in Yorubaland (Western Nigeria) [11].
Picralima nitida (Abere) has been used as
antimalarial, antifungal, analgesic and an-
tidiabetic agent. Traditionally, the seeds are
crushed or grinded into powdery form and tak-
en orally, for the treatment of malaria, diar-
rhoea, diabetic, as a painkiller and stimulant.
The antimicrobial, antipyretic and antifungal
activities of this plant have been reported [12–
14]. Pronounced inhibitory activities against
asexual erythrocytic forms of Plasmodium
falciparum was reported to be highest in the
root, stem, bark, and fruit rind extracts while
the leaf and seed extracts yielded much lower
activity or were completely inactive [15]. The
antimalarial activity of the plant has been at-
tributed to its alkaloid components [16–18].
The trypanocidal and antileshmanial activi-
ties of the plant have been reported [19, 20].
The plant also possesses opium analgesic [21,
22] and anticholinesterase properties [23]. The
seeds of Picralima nitida extract have also
been reported to possess hypoglycaemic, with
the coconut water extract of the seed having
higher hypoglycaemic effect than the aqueous
extract [24, 25].
This study was designed to evaluate some
possible untoward effects the P. nitida seed
extracts could have while using it as an hypo-
glycemic or antidiabetic agent using both bio-
chemical and histological tests.
Materials and methods
Materials
Fresh seeds of Picralima nitida were pur-
chased from Itoku market in Abeokuta, Ogun
State, Nigeria and were authenticated by Dr.
Aworinde (Plant taxonomist) of the Depart-
ment of Biological Sciences, Federal Universi-
ty of Agriculture Abeokuta. Technology of the
seeds obtaining is contain washing and rinsing
with distilled water, mopping with clean tis-
sue paper, cutting into pieces and allowed to
air dried after which they were grinded into
powdery form.
Animals
42 adult Wistar rats purchased from the
College of Veterinary Medicine, University
of Agriculture Abeokuta were used for this
study. The rats were allowed to acclimatize for
seven days in cages at room temperature. They
were given free access to commercial feed and
water ad libitum.
A Technology of Extract Preparation
Twenty-five grams (25 g) each of the dried
powdery form of the Picralima nitida seed was
soaked into 500 ml of distilled water, coconut
water and methanol separately. The mixture
was shaken vigorously and stirred at regular
interval for 48 hrs to allow for equilibration
of the mixture. The mixture was then filtered
using sterile muslin bag, and the filtrate ob-
tained was concentrated to constant residual
weight which was used to calculate percentage
yield. The extract obtained was kept inside air
BIOTECHNOLOGIA ACTA, V. 7, No 2, 2014
94
tight container and stored at 4 °C prior to use.
The percentage yield for methanolic, dis-
tilled water and coconut water extracts were
74.6%, 25.04% and 35.8% respectively.
Determination of antiradical activity of Pi-
cralima nitida
The antiradical activity of the plant was
determined by using 1, 1-diphenyl-2-picryl-
hydrazyl (DPPH) radical scavenging activity
as described by Ayoub et al., [26]. The DPPH
radical scavenging activity was calculated us-
ing the formula:
DPPH radical scavenging activity (%) =
[(A0 – A1/A0) × 100].
Where A0 is the absorbance of the control,
A1 is the absorbance of extract or standard
sample.
Induction of Diabetes Mellitus
Diabetes mellitus was induced in the rats
following the method described in [26]. The
rats were given a single intraperitoneal injec-
tion of 150 mg/kg of freshly prepared alloxan
monohydrate dissolved in normal saline after
an overnight fasting. Rats that survived after
the seventh day with blood glucose concentra-
tions more than 150 mg/100ml were consid-
ered diabetic.
Animal grouping and administration of extract
The rats were divided into seven groups of
six rats per group and treated as follows:
Group 1: served as control; non-induced
rats and were given free access to normal
feed and water ad libitum. Group 2: untreated
diabetes-induced rats. Group 3: non-induced
rats, received coconut water only. Group 4:
diabetes-induced rats, received coconut water
extract (100 mg/kg body weight). Group 5: dia-
betes-induced rats, received distilled water ex-
tract (100 mg/kg body weight). Group 6: dia-
betes-induced rats, received methanol extract
(100 mg/kg body weight). Group7: diabetes in-
duced rats, received 10 mg/kg body weight of
glibenclamide (standard drug).
Blood Glucose Determination
A drop of blood was collected by cutting the
tip of the tail of conscious rat and placed on the
strip of ACCU-CHEK glucometer for the de-
termination of blood glucose.
Periodic Weighing
Body weights of each rats in all the groups
was measured every three days throughout the
period of the treatment.
Blood collection and dissection
At the end of the experiment, the rats were
anesthesized under light diethyl ether, blood
samples was collected from each rat by cardi-
ac puncture into heparinised tubes. The tubes
were rocked gently to allow proper mixing of
the blood. Thereafter, the rats were dissected
and the liver and kidneys of each rat excised,
mopped, weighed and stored in 10% formalin
for histopathological studies.
Preparation of Plasma
Test tubes containing whole blood samples
collected were centrifuged at 4000 rpm for five
minutes, the supernatant (plasma) was decant-
ed into another tube and labeled accordingly.
Biochemical Analysis
Plasma Aspartate Aminotransferase (AST)
and Plasma Alanine Aminotransferase (ALT)
assays: AST and ALT was measured spectro-
photometrically by the method of Reitman and
Frankel [27] as described in Randox kit manual.
Plasma Urea
Urea in the plasma was determined using
Urease-Berthelot method as described in Ran-
dox kit manual [28].
Plasma Cholesterol
Plasma cholesterol was determined after
enzymatic hydrolysis and oxidation according
to the method described by Trinder [29] as de-
scribed in Randox diagnostic kit manual.
Plasma Triglycerides
Plasma triglycerides are determined after
enzymatic hydrolysis with lipases according to
the method described in Randox kit manual.
Estimation of vLDL-Cholesterol
The concentration of Very Low Density Li-
poprotein (vLDL) cholesterol was calculated by
modification of the Friedewald formular [30].
vLDL-Cholesterol was calculated as Triglycer-
ol concentration obtained divided by five.
Plasma Gamma-glutamyltransferase (GGT)
Plasma GGT was measured spectrophoto-
metrically by the method described by Szasz
[31] as described in Randox Diagnostic kit
manual.
Lipid peroxidation
The extent of lipid peroxidation was esti-
mated in terms of thiobarbituric acid reactive
species (TBARS) by measuring the amount of
malondialdehyde (MDA) formed according to
the method described by Drapper et al. [32],
with slight modifications.
Determination of plasma sodium and po-
tassium
Plasma sodium and potassium concentra-
tion were determined using the flame photom-
etry method.
Determination of Bicarbonate concentration
The plasma bicarbonate concentration was
determined by titration.
Experimental articles
95
Packed Cell Volume (PCV) Estimation
PCV was determined by the microhaemato-
crit method [33] as described by Omotainse and
Anosa [34].
Haemoglobin (Hb) Concentration
The haemoglobin concentration was deter-
mined colorimetrically using Cypress diagnos-
tic kit.
Red Blood Cell (RBC) and Whole Blood Cell
(WBC) count
The RBC and total WBC counts were car-
ried out by the use of the Neubauer haemocy-
tometer according to the method of Schalm et
al. [33] as described by [34].
Histopathological examinations
Histopathological examinations of the
liver, kidney and pancreas was done following
the standard Heamatoxylin and Eosin (H&E)
staining method as described by Krause [35].
Statistical Analysis
Results obtained were expressed as mean
± S.D. The levels of homogeneity among the
groups were tested using one-way Analysis
of Variance (ANOVA) and Duncan Multiple
Range Test (DMRT). Analysis was done using
statistical package for Social Science (SPSS)
version 16.0. Difference at P < 0.05 were con-
sidered statistically significant.
Results and discussion
The antiradical activity of the different ex-
tracts of Picralima nitida seeds
The antiradical activity of all the studied
extract of P. nitida seed are summarized in
Table 1. The methanolic extraction of the seed
has the lowest absorbance among the three
when compared with the standard, indicating
higher free radical scavenging activity.
Body weight of Animals
Table 2 shows the summary of the effect
of the plant seed extracts on body weights of
the animals. After the intraperitoneal injec-
tion of 150 mg/kg alloxan monohydrate, mean
body weight of all the alloxan-induced groups
were significantly reduced (P < 0.05). During
treatment, all diabetic-induced but treated
rats showed increase (P < 0.05) in body weight
when compared with their weights after induc-
tion prior treatment. The result showed higher
weight gain in the rats treated with methano-
lic and coconut water extract of P. nitida seeds
when compared to other treated groups. There
was consistent increase in weight of the nor-
mal control groups while the diabetes induced
but not treated group showed a consistent de-
crease in weight throughout the experiment.
The summary of the blood glucose levels of
the rats is presented in Table 3. Before induc-
tion of diabetes the mean blood glucose level
of the rats ranged from 81.67 to 91.33 mg/dl.
There was increase in the mean blood glucose
of all the induced groups, having values be-
tween 223.33 and 265.33 mg/dl which con-
firmed them diabetic. All the groups treated
with the extracts had decreased mean glucose
values respectively. Glibenclamide treated
rats also showed decrease in mean blood glu-
cose from 245.00 to 110.67 mg/dl at the end
of the experiment. However, the diabetic un-
treated rats maintained high plasma glucose.
Effects of treatment on the haematologi-
cal parameters are presented in table 4. The
Packed Cell Volume (PCV) and Hb contents of
the alloxan-induced but untreated group and
the induced but distilled water extract treated
group were significantly lower (P < 0.05) than
the control. Only the induced but methanol ex-
tract treated group had no significant differ-
ence in the RBC content when compared with
the control. The WBC and lymphocyte con-
tents of all the alloxan induced groups were
significantly lower (P < 0.05) than that of the
control, with the alloxan-induced untreated
group having the lowest value. Comparison
the lymphocyte contents of the induced groups
with the non-induced group, only the alloxan-
induced and the glibenclamide-treated groups
had a significant increase (P < 0.05). Alloxan
induced-methanolic extract-treated group had
the least lymphocyte level.
Table 5 depicts the effect of treatments
on some lipid profile parameters. The re-
sult showed that alloxan caused significant
(P < 0.05) increase in the lipid profile (total
Table 1. The antiradical activity of the different
extracts of Picralima nitida seeds
EXTRACT ABSOR BANCE % INHIBI TION
AQUEOUS
EXTRACT 0.444 19.36
METHA NOLIC
EXTRACT 0.348 36.73
COCONUT
WATER
EXTRACT 0.528 4.09
STANDARD
(VIT C) 0.0015 100
BIOTECHNOLOGIA ACTA, V. 7, No 2, 2014
96
Table 2. Effect of treatments of Picralima nitida seeds extract on body weight (g) of rats
GRP TREAT-
MENT DAY 1 DAY 7 DAY 10 DAY 13 DAY 16 DAY 19
1Control 80 ±11.18a99 ±13.42a107±10.37b117±10.37b122 ±7.58bc 132±10.37bc
2Alloxan
only 129 ±4.18b118±4.48b115 ±4.08b111 ±7.50b109 ±9.02a108±11.68ab
3CW only 80 ±11.18a103 ±13.0ab 115±11.18b128±13.51bc 143±11.51cd 151 ±15.58c
4A + CW 120±20.92b104±11.94ab 104±20.43ab 111 ±22.75b123±16.47bc 134 ±13.77c
5A + DS 105±20.92b92 ±16.05a90 ±13.69a88±9.75a97 ±10.41a98 ±17.56a
6A + MET 124±6.88b115±10.61b123±18.37b142±5.77c147±10.41d150±10.00c
7A + GLIB 126±29.88b109±20.43ab 123±16.05b127±23.63bc 132±27.54bcd 142±27.54c
Notes: hereinafter values are mean ± SD. Values within the same row with different superscripts are signifi-
cantly different at P < 0.05, (n = 6); CW only: Coconut water only; A + CW: Alloxan + Coconut water extract of
P. nitida seed; A + DS: Alloxan + Distilled water extract of P. nitida seed; A + MET: Alloxan + Methanol extract
of P. nitida seed; A + GLIB: Alloxan + Glibenclamide.
Table 3. Effect of treatments of Picralima nitida seeds on the plasma glucose level (mg/dL) of rats
GR TREAT-
MENT DAY 1 DAY 7 DAY 10 DAY 13 DAY 16 DAY 19
1Control 90.60±5.18a84.80±3.96a85.20±5.26a81.6±6.27a84.6±4.22a81.4±5.94ab
2Alloxan
only 91.33±6.51a265.33±41.59b292±66.73c250.67±35.02c249.67±40e247.67±46.2d
3CW only 83.60±5.59a82.80±5.07a 83±4.47a79.6±8.68a91.8±1.3a75±9.03a
4A+ CW 86.75±10.99a234.25±32.09b227.75±31.17b204±9.35b194.75±8.66d144.5±45.2c
5A + DS 81.67±7.57a223.33±29.19b287±62.64c223.33±34.5bc 174±3.46cd 128.67±21c
6A+MET 88.67±3.06a249.33±39.53b214.67±11.72b196.33±29.02b150.67±5.13bc 122.67±6.11bc
7A+GLIB 79.33±14.01a245.00±33.45b203.33±39.07b193±29.51b134.67±22.37b110.6±4.58abc
Table 4. Effect of treatments of Picralima nitida on some hematological parameters of rats
GRP TREAT-
MENT PCV (%) Hb (g/dl) RBC
(× 106/mm3)WBC
(×103 mm3)PLAT
(×103 mm3)LYMP (%)
1Control 45.08±3.03cd 15.05± 0.89c7.66±0.48d10.10±0.69d638.20±7.86e64.00±2.55ab
2Alloxan
only 31.53±1.67a10.34±0.43a5.43±0.31a5.13±0.21a217.00±6.25a77.67±3.51d
3CW only 41.40±2.69bc 13.99±0.78bc 7.30±0.29cd 9.04±0.27d637.40±7.89e65.20±2.28b
4A + CW 41.08±0.72bc 13.69±0.59bc 6.88±0.21bc 7.68±0.33c244.67±8.60c62.75±2.87ab
5A + DS 38.40±6.27b12.74±2.08b6.43±0.47b6.47±0.40c294.67±8.33c63.00±3.61ab
6A + MET 46.70±1.45d15.24±0.52c7.80±0.26d8.33±0.45c419.33±6.03d60.00±3.46a
7A + GLIB 42.73±2.18bc 14.20±0.63bc 7.03±0.47bc 5.80±0.36ab 228.00±11.14a71.33±1.53c
Experimental articles
97
cholesterol, triglyceride and vLDL) of the
treated and untreated groups when compared
with the control. However, treatments with
the P nitida seed extracts showed significant
(P < 0.05) reduction of the lipid profile levels.
The cholesterol, triglycerol, and very Low Den-
sity Lipoprotein values of the alloxan-induced
glibenclamide treated group were significant-
ly lowered (P < 0.005) when compared with
the control. The alloxan-induced but distilled
water treated group showed a significant in-
crease in the lipid profile when compared with
all other groups.
The influence of treatments on some liver
function test parameters is presented in Table 6.
The alloxan- induced but untreated group
showed a significant increase in the activities
of all marker enzymes namely; GGT, AST, and
ALT, when compared with the control. None of
the treatments was able to bring the activities
of these enzymes to the level of the control.
Electrolytes and urea plasma levels of
the different groups of rats were presented
in Table 7. There was no significant differ-
ent (P > 0.05) between distilled water extract
treated group and glibenclamide treated group
as well as between the control and group ad-
ministered coconut water only. All the renal
function parameters (urea, sodium, potassium
and bicarbonate) of the glibenclamide group
reduced significantly (P < 0.05) when com-
pared with the control group.
Table 8 shows the summary of values of MDA
concentration in the plasma of different groups.
The MDA content of the extract treated groups
decreased (P < 0.05) when compared to the con-
trol. However, the group administered alloxan
only had a significant increase (P < 0.05) in the
MDA content when compared to the control.
HISTOLOGICAL STUDIES
The liver
Histopathological examination of the liver
of the rats in groups 1 and 3 showed normal ap-
pearance/architecture of the hepatocytes, bile
ducts and blood vessels (Fig. 1).
Examination of the liver of untreated al-
loxan induced diabetic group showed lesions
which include mononuclear cell infiltration
of the portal triads particularly around the
bile duct (Fig. 2), necrosis of the hepatocyte,
diffuse disorganisation of the hepatocyte
cords and diffuse distribution of mononuclear
cells in the sinusoids. Photomicrograph of the
treated groups also showed varying degree of
lesions (Fig. 2–7). The degree of lesions in the
treated groups are however milder compared
to the untreated group.
Table 5. Effect of treatments on some lipid profile parameters of rats
GRP TREATMENT Total CHOL (mg/dl) TRIG (mg/dl) vLDL (mg/dl)
1Control 87.20±3.69a62.40±3.49ab 12.48±0.69ab
2Alloxan only 120.47±2.99e96.70±2.21d19.34±0.44d
3CW only 87.72±3.05a64.80±2.89ab 12.88±0.52ab
4A + CW 97.88±1.42b67.03±7.55bc13.41±1.51bc
5A + DS 110.67±3.16d74.40±4.79c14.88±0.96c
6A + MET 103.63±4.30c67.23±5.39bc 13.45±1.08bc
7A + GLIB 98.03±2.85b58.50±5.99a11.70±1.19a
Table 6. Effect of treatments on activities of some markers enzymes of hepatotoxicity in rats
GRP TREATMENT GGT (U/L) AST (U/l) ALT(U/l)
1Control 23.82 ± 2.10a22.65 ± 2.90a37.02±2.20a
2Alloxan only 67.10 ± 3.30e79.01 ± 0.72f54.38±4.45d
3CW only 25.00 ± 1.49a35.93 ± 1.82b37.76±1.19a
4A + CW 34.38 ± 2.11bc 45.16 ± 4.83c38.78±1.75ab
5A + DS 38.90 ± 1.92d55.17 ± 4.81d40.63±1.31ab
6A + MET 35.40 ± 3.48cd 37.80 ± 0.70b42.00±1.76bc
7A + GLIB 30.97 ± 3.29b60.69 ± 1.54e45.67±3.79c
BIOTECHNOLOGIA ACTA, V. 7, No 2, 2014
98
Table 7. Effect of treatments of Picralima nitida seeds on renal function parameters in rats
GRP TRTMENT UREA
(mg/dl)
SODIUM
(mmol/L)
POTASSIUM
(mmol/L)
BICARBONATE
(mmol/L)
1Control 19.58±1.86a73.00±7.42a3.56±0.61a12.00±1.23a
2Alloxan only 54.43±4.15e131.67±3.50d9.53±0.31d22.33±1.16d
3CW only 19.64±2.29a86.40±1.67b7.02±0.33bc 16.00±1.00b
4A + CW 44.05±1.82d94.75±12.37bc 7.13±0.61bc 16.50±2.08bc
5A + DS 35.43±2.45c98.67±9.29c7.83±0.72c18.33±1.16c
6A + MET 26.97±1.06b104.33±3.22c8.07±0.59c18.00±1.00bc
7A + GLIB 34.93±1.59c85.00±2.00b6.10±2.23b13.67±0.58a
Table 8. Effect of treatments of Picralima nitida
seeds on lipid peroxidation level in rats
GRP TREATMENT MDA (µmol/L)
1Control 0.48±0.33a
2Alloxan only 3.20±0.76c
3CW only 0.49±0.36a
4A + CW 1.24±0.38b
5A + DS 1.25±0.27b
6A + MET 1.27±0.21b
7A + GLIB 1.19±1.91b
Fig. 1. Photomicrograph of the liver section
showing normal appearance of the perivascular
spaces of the hepatocytes (P) and blood vessel (V)
in the liver:
of the normal control groups (1 and 3). ×200 H&E
P
Fig. 2. Photomicrograph of the liver section
showing infiltration of perivascular space (V)
of the hepatocytes:
in the untreated diabetes induced group.
×200 H&E V
Fig. 3. Photomicrograph of the liver section show-
ing bile duct hyperplasia (D) in the hepatocytes:
of the induced groups treated with 100mg/kg
of coconut water and distilled water extraction
of P. nitida. ×400 H&E
D
Fig. 4. Photomicrograph of the liver section show-
ing perivascular cellular infiltration (PI) and dis-
organized hepatic cords in the hepatocytes (DH):
of the diabetic group treated with glibenclamide.
×400 H&E
Fig. 5. Photomicrograph of the liver section show-
ing severe periportal cellular infiltration (P) and
disorganized hepatic cords of the hepatocytes:
in the diabetic group treated with coconut water
and methanolic extraction of P. nitida seeds.
×200 H&E
V
P
V
P
D
DH
Experimental articles
99
The Kidney
Photomicrograph of the kidney of the con-
trol rats revealed normal appearance of the
glomeruli, tubules and blood vessels (Fig. 8).
Histopathological examination of the un-
treated group showed necrosis of the Bow-
man’s capsule, enlarged perivascular space
(oedema) of the kidney and tubular necrosis
of the kidney. The group treated with coconut
water extraction showed diffuse massive tu-
bular epithelial cell degeneration, glomerular
necrosis, congestion of the vessels and degen-
eration of the renal tubules. Similar lesions or
more severe than these were seen in the other
treated groups.
The Pancreas
The lesions observed in the different
groups in the pancreas were as summarized in
the Table 9 and Figures 11–18. These lesions
include necrosis of the exocrine and fat cells,
enlargement of the islet of Langerhans, hyper-
plastic ducts, proliferation of the blood vessels
and blockage of the tubules by necrotic and
inflammatory cells. Other lesions include the
thickening of the tubular wall, perivascular
necrosis and fibrosis as well as vascular degen-
eration of the cells of the islets.
Diabetes is a common and very prevalent
disease affecting the people in both developed
and developing countries [36]. According to
the World Health organization’s report in
2006, at least 171 million people worldwide
suffer from diabetes [37]. The incidence of
diabetes mellitus is increasing rapidly and it
is estimated that by the year 2030, this num-
ber will double [37]. The primary goal of treat-
ment of diabetes is to bring the elevated blood
sugars down to a normal range in order to al-
leviate or prevent complications.
The use of herbal products for medicinal
benefits has played an important role in near-
ly every culture on earth and for many years,
not only because they are relatively cheap and
readily available, but also due to ease with
Fig. 1. Photomicrograph of the liver section
showing normal appearance of the perivascular
spaces of the hepatocytes (P) and blood vessel (V)
in the liver:
of the normal control groups (1 and 3). ×200 H&E
P
Fig. 2. Photomicrograph of the liver section
showing infiltration of perivascular space (V)
of the hepatocytes:
in the untreated diabetes induced group.
×200 H&E V
Fig. 3. Photomicrograph of the liver section show-
ing bile duct hyperplasia (D) in the hepatocytes:
of the induced groups treated with 100mg/kg
of coconut water and distilled water extraction
of P. nitida. ×400 H&E
D
Fig. 6. Photomicrograph of the liver section show-
ing severe necrosis of hepatocytes (N):
in the group treated with methanolic extraction
of P. nitida seeds. ×200 H&E
Fig. 7. Photomicrograph of the liver section
showing necrosis of the hepatocytes (NH).
×200 H&E
Fig. 8. Photomicrograph of a section of the kidney
of control group:
showing the normal appearance of the glomerulus
(G), tubules (T), and blood vessels of the kidney.
×200 H&E
Fig. 9. Photomicrograph of a section of the kidney:
showing necrotic glomerulus (NG) of the diabetic
groups treated with coconut water, distilled water
and methanolic extraction of the seeds of P. nitida.
×400 H&E
Fig. 10. Photomicrograph of a section of kidney
showing congested blood vessels (B):
in the groups treated with coconut water, distilled
water, methanolic extraction of P. nitida seeds,
also in glibenclamide treated group. ×200 H&E
N
NG
B
NH
G
T
BIOTECHNOLOGIA ACTA, V. 7, No 2, 2014
100
Table 9. Summary of the severity of lesions in the pancreas in the various treatment groups
LESIONS GROUPS
1 2 3 4 5 6 7
Necrosis of exocrine and fat cells ++ + + + +
Enlarged islet of langerhans ++ +++ +++ +++ +
Hyperplastic ducts and ductular proliferation +++ +++ ++ +
Blockage of ductular lumen by inflammatory cells ++ + ++ ++ ++
Thickening of the walls of the tubules and vessels ++ +++ ++ +
Peri-vascular necrosis and fibrosis ++ +++ ++ ++
Vascular degeneration of the cells of the islets and
fat cells – – – + + +++ +
Notes: – not present; + mild; ++ moderate; +++ severe.
Fig. 11. Photomicrograph of the pancreas section:
showing of normal appearance of the langerhans
isletsf (IL) in the pancreas of normal control
groups. ×400 H&E
Fig. 12. Photomicrograph of a section
of the pancreas:
showing degeneration of the langerhans islets
(DI) and thickened duct (TD). ×100 H&E
PANCREAS: hyperplastic ductular wall
Fig. 13. Photomicrograph of a section of the pancreas:
showing mild hyperplasia (MH) of the ductular
wall. ×100 H&E
Fig. 14. Photomicrograph of a section of the pancreas:
showing severe hyperplasia (SH) of the ductular
wall. ×400 H&E
Fig. 15. Photomicrograph of a section of the pancreas:
showing mild vacuolation (MV) of islet cells.
×200 H&E
Fig. 16. Photomicrograph of a section of the pancreas:
showing periductal cellular infiltration (PI).
×200H&E
IL
DI
TD
MH PI
MV
SH
Experimental articles
101
which they can be administered. One of such
disease condition in which herbal products is
being advocated for it management is diabetes
mellitus [38].
Changes in body weight are usually consid-
ered in all physiological conditions. The dia-
betic control group showed a remarkable de-
crease in body weight at the end of the study.
On the other hand, increase in body weight was
observed in all the seed extract treated groups
and also in the control groups.
Increased oxidative reactions level shows
the production of free radicals and reactive
oxygen species which are formed under nor-
mal physiological conditions but may become
deleterious when they are not quenched by the
antioxidant molecule(s) within the systems
[39]. The antiradical activity of the different
extracts of Picralima nitida seeds showed that
the methanolic extract of the seed have the
highest percentage inhibition 36.73%, thus
having higher free radical scavenging activ-
ity, this is followed by the aqueous extract
19.36%, while the coconut water have the
least percentage inhibition of 4.09%.
There have been reports on the increase
in blood glucose during diabetes in rats and
humans [40–44]. The blood glucose lowering
effect of P. nitida extract after induction by
alloxan is in agreement with the reports of Sa-
lihu et al. [24] and Inya-Agha et al. [45]. This
could be attributed to the presence of some ac-
tive ingredient(s) that enhances its ability to
increase the permeability of cell plasma mem-
brane to glucose. There has been an increase in
the prevalence of atherosclerosis and hyperlip-
idaemia among diabetics worldwide [46]. This
is said to occur as a result of the altered lipid
profile in the serum of diabetic patients [47,
48], which is as a result of increase in plasma
triglycerides and total cholesterol levels. Hy-
percholesterolemia has been reported to oc-
cur in alloxan-diabetic rats [49, 50]. The most
common lipid abnormalities in diabetes are hy-
pertriglyceridemia and hypercholesterolemia
[51]. This was observed in the lipid profile of
the diabetic control group. In our study there
was a significant decrease in the lipid profiles
of all the three groups treated with extracts
of Picralima nitida seeds. However, the group
treated with coconut water seed extract showed
a significant decrease in total cholesterol level
when compared with the control, followed by
the methanolic extract treated group. There
was also a significant decrease in the total
triglyceride and vLDL of all the seed extract
treated groups. However, glibenclamide been
a standard drug for diabetes showed a most
pronounced decrease in the lipid profile levels.
The observed hypolipidemic effect may be at-
tributed to decreased blood glucose levels and
decreased cholesterogenesis as well as fatty
acid synthesis. This similar to the report of
Bopanna et al. [52].
Anaemia is a medical condition caused by
an abnormally low number of RBC, PCV and
the Hb content. The anaemia occurring in di-
abetes is due to the increased non-enzymatic
glycosylation of RBC membrane protein which
correlates with hyperglycemia [1]. This study
showed a marked reduction in the PCV and Hb
content in the diabetes induced but untreated
group. The treated groups showed improve-
ment in the PCV and Hb content. This indi-
cates that the seed extract of Picralima nitida
might probably contain some phytochemicals
that were able to increase or boast the level of
packed cell volume and Hb concentration.
Aspartate- (AST) and alanine transaminase
(ALT) are found predominantly in the liver,
and are biochemical markers for liver injury
in patients [3]. Elevated activity of both en-
zymes above normal is an indication of possible
liver damage. Although, the diabetes induced
groups all had high activities of AST and ALT
when compared with the control, there was
Fig. 17. Photomicrograph of a section of the pancreas:
showing necrotic islet (NI). ×400H&E
Fig. 18. Photomicrograph of a section of the pancreas:
showing thickened pancreatic duct. ×200 H&E
NI
TD
BIOTECHNOLOGIA ACTA, V. 7, No 2, 2014
102
significant reduction in the activities of these
enzymes in all the extract treated groups. Only
the untreated group had very high levels, thus
indicating liver impairment.
Plasma GGT is a measure of the hepatobili-
ary system. Elevated plasma GGT activity has
been reported to be found in diseases of the liv-
er, biliary system and pancreas [53]. There was
a significant reduction in the plasma GGT of
the treated groups. This indicates an improve-
ment in the hepatobilary system.
High lipid peroxidation level is a sign of
oxidative damages. Oxygen free radicals spe-
cies have been implicated in the pathogenesis
of diabetes mellitus [54]. This was seen in all
the diabetes — induced groups, but the extract
treated groups showed significant decrease in
the extent of lipid peroxidation. This could
probably be due to the presence of the alkaloids
and glycosides present in the seed extracts.
However, the untreated group had a very high
level lipid peroxidation.
Picralima nitida seeds have been reported
to be rich in alkaloids and glycosides [8, 24].
Glycosides present in the seeds have been re-
ported to be responsible for the blood glucose
reduction through pancreatic and extra pan-
creatic effect. These pancreatic and extra pan-
creatic effects on the blood glucose could be
through prevention of hepatic glucose over
production, increase in glucose uptake by the
muscles, inhibition of gastric emptying and/or
increase in glucose permeability of plasma cell
membrane.
Although, diabetes mellitus have been re-
ported to be associated with varied histologi-
cal changes in different organs, in this study,
histological examinations of the pancreas of
the treated and untreated groups showed vary-
ing degree of degenerations but the extent of
severity in the lesions were more pronounced
in the extract treated groups. The histopatho-
logical examination of the liver and kidney
also revealed varying degree of degenerations
in these organs. This suggests the inability of
the different extracts administered to, either
prevent the uptake of alloxan into the ü cells,
or to interfere with its destructive actions in
the cells. It could therefore be concluded that
although, Picralima nitida seed extracts were
seen to reduce diabetes to some extent (with
the methanolic extract having highest anti-
oxidant property than the others), however,
this plant seed extract did not produce any ob-
servable ameliorative or regenerative proper-
ties to the organs damaged by alloxan. Thus,
caution should be exercised while taking P.
nitida seed extracts decoctions (tradition-
ally) or recommending it as a possible remedy
against diabetes.
REFERENCES
1. Umar I.A., Mohammed A., Dawud F. A.,
Kyari, H., Abdullahi M. Anti-diabetic action
of the aqueous extract of Ocimum suave in al-
loxan induced diabetic rats. Afr. J. Biotech-
nol. 2012 11(38), 9243–9247.
2. Lubert S.. Metabolic derangement in diabetes
result from relative insulin insufficient and
glucogen excess. Intergration of metabolism.
Biochemistry. 1995, 4th ed., P. 780.
3. Harris M. National Diabetes: Data Group Na-
tional institutes of health. Diabetes and Diges-
tive and Kidney Diseases. «Diabetes in Ameri-
can». 2nd ed. NIH. 1995, P. 1395–1468.
4. Meyer P. A. Digestion and absorption in: Mur-
ry R. K. Harper’s Biochemistry Stamford. Ap-
pleton and Lange. 2000, P. 178–188.
5. Barnett H. A., O’ Gara G. Diabetes and the
heart: clinical practice series. Churchhill liv-
ingstone Edinburge, United Kingdom. 2003,
P. 7–30.
6. Harris M. I., Flegal, K. M., Cowie C. C., Eber-
hardt M. S., Goldstein D. E., Little R. R., Wied-
meyer H. M., Byrd-Holt D. D. Prevalence of di-
abetes, impaired fasting glucose, and impaired
glucose tolerance in United States adults. The
third National Health and Nutrition Examina-
tion Survey, 1988–1994. Diabetes care. 1998,
21(4), P. 518–524.
7. Nathan D. M., Cleary P. A., Backlund, J. Y., Ge-
nuth S. M., Lachin J. M., Orchard T. J., Ras kin
P., Zinman B. Intensive diabetes treatment and
cardiovascular diseases in patient with type 1
diabetes. New England J. Med. 2005, 353(25),
P. 2643–2653.
8. Corbett A. D., Menzies J. R. W., Macdonald, A.,
Pa ter son, S. J., Duwiejua, M. The opioid ac-
tivity of akuammine, akuammicine and
akuammidine: alkaloids from Picralima nitida
(fam. Apocynaceae). Brit. J. Pharmacol. 1996,
V. 119, P. 334.
9. Ubulom P., Akpabio E., Udobi C., Mbon R. An-
tifungal activity of aqueous and ethanolic
extracts of picralima nitida seeds on Aspergil-
lus flavus, Candida albican and Microsporum
canis. Pharmaceut. Biotechnol. 2011, 3(5),
P. 57–60.
10. Keay R. W. J., Onochie C.I.A., Stemfield D. D.
Nigerian Trees. Federal Department of Forest
Resources: Ibadan, Nigeria. 1964, P. 1–20.
11. Aguwa C. N., Ukwe C. V., Inya-Agha S. I.,
Okonta J. M. Antidiabetic effect of Picralima
Experimental articles
103
nitida aqueous seed extract in experimental
rabbit model, J. Natl. Remed. 2001, V. 1, P.
135–139.
12. Iroegbu C. U., Nkere C. K. Evaluation of the
antibacterial properties of Picralima nitida
stem bark extracts. Intern. J. Mol. Med. Adv.
Sci. 2005, 1(2), P. 182–189.
13. Ansa-Asamoah R., Kapadia G. J., Lloyed H.
A., Sokoski E. A. Picratidine, a new indole al-
kaloid from Picralima nitida seeds. J. Nat.
Prodts-Lloydia. 1990, V. 5, P. 975–977.
14. Magihu M., Mbuyi M., Ndele M. B. Use of
Medicinal Plants among the Pygmes (Mbute)
to treat Malaria in the area of Mambasa, Itu-
ri, Zaire. The Biodiversity of African Plants,
Wageningen, Netherland. 1996, P. 741–746.
15. Francois G., Ake Assi L., Holenz J., Bring-
mann G. Constituents of Picralima nitida
display inhibitory activities against asexual
erythrocytic forms of Plasmodium falci-
parum in vitro. J. Ethnopharmacol. 1996,
V. 54. P.113–117.
16. Iwu M. M., Klayman D. L. Evaluation of in vi-
tro antimalarial activity of Picralima nitida
extracts. J. Ethnopharmacol. 1992, V. 36, P.
133–135.
17. Kspadia G. J., Angerhofer C. K., Ansa-Asa-
moah R. Akuammine: an antimalarial in-
dolemonoterpene alkaloid of Picralima nitida
seeds. Planta Medica. 1993,59(6), P. 565–566.
18. Moeller B. L., Seedorff L., Nartey F. Alka-
loids of Picralima nitida. Phytochemistry.
1972, V.11, P. 2620–2621.
19. Iwu M. M, Klayman D. L, Bass G. T. Antima-
larial activity of Indole alkaloids from Pic-
ralima nitida. Am. J. Trop. Med. Hyg. 1992,
V. 47. P. 179–186.
20. Wosu L. O., Ibe C. C . Use of extract of Pic-
ralima nitida bark in the treatment of ex-
perimental trypanosoniasis: A preliminary
study. J. Ethnopharmacol. 1989, V. 25,
P. 263–268.
21. Menzies J. R. W., Paterson S. J., Duwiejua M.,
Corbelt A. D. Opioid activity of alkaloids ex-
tracted from Picralima nitida (fam. Apocy-
naceae). Eur. J. Pharmacol. 1998, V. 350,
P. 101–108.
22. Ezeamuzie I. C., Ojinnaka M. C., Uzogara E. O.,
Oji S. E. Antiinflammatory, antipyretic and
antimalarial activities of a West African me-
dicinal plant Picralima nitida. Afr. J. Med.
Med. Sci. 1994, 23(1), 85–90.
23. Levy A., Collin M. C. Anticholinestaric prop-
erties of pseudo Akamminigine alkaloid of
Picralima nitida Apocynaceae. Ann. Pharma-
ceut. Franc. 1978, V. 36, P. 77–83.
24. Salihu M. A., Luqman A. O., Oshiba O. J. Rabiu
O. Jimoh, Sikiru A. Jimoh, Ayokunle Olawepo,
Adesola I. R. Abioye. Comparative study of
the hypoglycemic effects of coconut water
extract of Picralima nitida seeds (Apocyna-
ceae) and Daonil in alloxan-induced diabetic
albino rats. Afr. J. Biotechnol. 2009, 8(4), P.
574–576.
25. Okonta J. M., Adibe M. O., Ubaka C. M. Antiulcer
activity of methanolic extract and fractions of Pi-
cralima nitida seeds (Apocynacaea) in rats. Asian
Pacific J. Trop. Med. 2010, V. 7, P. 13–15.
26. Ayoub R. C., Zahid M. Q., Syed A. R., J. Wil-
liam, M. Arshad. Quantitative determination
of Antioxidant potential of Artemisia persica.
Analele Universitatll din Bucuresti-Chimie
(serie noua). 2010, 19(1), P. 23–30.
27. Reitman S., Frankel S. A colorimetric method
for the determination of serum glutamic oxa-
loacetic and glutamic pyruvic transaminase.
Amer. J. Clin. Pathol. 1957, V. 28, P. 56–63.
28. Weatherburn M. W. Phenol-hypochlorite re-
action for determination of ammonia. Anal.
Chem. 1967, V. 39, P. 971–982.
29. Trinder P. Mono-reagent enzymatic glucose.
Clinical Chemistry W. B. Sanders, Philadel-
phia. 1969. P. 24–27.
30. Sandkamp M., Funke, H., Schulter Köhler E,
Assmann G. Lipoprotein (a) is an indepen-
dent risk factor for myocardial infarction at a
young age. Clin. Chem. 1990, V. 36. P. 20–23.
31. Szasz G. Methods of Enzymatic Analysis. 2nd
English ed. New York, Academic Press Inc.
1974, P. 717–720.
32. Drapper H. H., Squines E. J., Mahemedi H. W.
A comparative elevation of thiobarbituric
acid method for the determination of malo-
ndialdehyde in biological material. Freemed,
1993, V. 15, P. 353–363.
33. Schalm O. W., Caroll E. J. In: Veterinary He-
matology. 3rd ed., Lea and Febiger, Philadel-
phia. 1975, P. 204–206.
34. Omotainse S. O., Anosa, V. O. Erythrocyte re-
sponse to Trypanosoma brucei in experimen-
tally infected dogs. Rev. Elev. Med. Vet. Pays
Trop. 1992, 45(3), P. 279–283.
35. Krause W. J. The art of examining and inter-
preting histological preparations. A student
handbook. Parthenon Publishing group, U. K.
2001, P. 9–10.
36. Erasto P., Adebola P. O., Grierson D. S., Afo la-
yan A. J. An ethanobotanical study of plants
used for the treatment of diabetes in the
Eastern Cape Province, South Africa. Afr. J.
Biotechnol. 2005, 4(2), P. 1458–1460.
37. American Diabetes Association. Total preva-
lence of diabetes and pre-diabetes. American
Diabetes Association. 2005, P. 15.
38. Osinubi A. A., Ajayi O. G., Adesiyun A. E.
Evaluation of the antidiabetic effect of aque-
ous leaf extracts of Tripinanthus butungil in
male spragne Dawley rats. Med. J. Islamic
World Acad. Sci. 2006, 16(1), P. 41–47.
39. Fang Y. Z., Yang S., Wu G. Free radical, an-
tioxidant and nutrition. Nutrition. 2002,
V. 18. P. 872–890.
BIOTECHNOLOGIA ACTA, V. 7, No 2, 2014
104
40. Ozougwu J. C. Anti-diabetic effects of Allium
cepa (onions) aqueous extracts on alloxan-
induced diabetic Rattus novergicus. J. Med.
Plants Res. 2010, 5(7), P. 1134–1139.
41. Hamme H. R., Martins S., Federlin K., Geisen K.,
Brownlee M. Aminoguanidine treatment in-
hibits the development of experimental dia-
betes retinopathy. Proc. Natl. Acad. Sci. USA.
1991, V. 88, P. 11555–11558.
42. Sharpe P. C., Yue K. M., Catterwood M. A.,
McMaster D., Trimble E. R. The effects of
glucose induced oxidative stress on growth
and extracellular matrix gene expression of
vascular smooth muscle cells. Diabetologia.
1998, V. 41, P. 1210–1219.
43. Tukuneu N. B., Bcyraktar M., Varli K. Rever-
sal of defective nerve conductions with vi-
tamin E supplementation in type 2 diabetes.
Diabetes Care. 1998. V. 21, P. 1915–1918.
44. Bell B. M., Hayes J. R., Stout R.W. Lipoprotein,
insulin and glycaemic control in diabetes. Hor-
mon. Metab. Res. 1984, V. 16, P. 252–260.
45. Inya-Agha S. I., Ezea S. C. Odukoya O. A.
Evaluation of picralima nitida: Hypoglycemic
Activity, Toxicity and Analytical Standards.
Intern. J. Pharmacol. 2006, 2(5), P. 576–580.
46. Pepato M. T., Baviera A. M., Vendramini R. C.
Perez Mda P., Kettelhut Ido C., Brunetti I. L.
Cissus sicyoides (Princess Vine) in the long-
term treatment of streptozotocin-diabetic
rats. Biotechnol. Appl. Biochem. 2003, V. 3,
P. 15–20.
47. Orchard T. J. Dyslipoproteinemia and dia-
betes. Endocrinol. Metab. Clin. North. Am.
1990, V. 19, P. 361–379.
48. Betteridge D. J. Diabetic dyslipidaemia. Am.
J. Med. 1994. V. 96, P. 25–31.
49. Sharma S. R., Dwivedi S. K., Swarup D. Hypo-
glycaemic and hypolipidaemic effects of Cin-
namon tomala nees leaves. Ind. J. Exp. Biol.
1996, V. 34, P. 372–374.
50. Pushparaj P., Tan C. H., Tan B. K. Effect of
Averrhoa bilimli leaf extract on blood glucose
and lipids in streptozotocin diabetic rats. J.
Ethnopharmacol. 2000, V. 72, P. 69–76.
51. Mitra K. S., Gopumadahavan S., Muralid-
har S. T., Anturlikar S. D., Sujatha M. B.
Effects of D-400, a herbomineral preparation
on lipid profile, hemoglobin and glucose
tolerance in streptozotocin induced diabetes in
rats. Ind. J. Exp. Biol. 1995, V. 33, P. 798–800.
52. Bopanna N. K., Kannan J., Gadgil S. Antidia-
betic and antihyperglycemic effects of neem
seed kernel powder on alloxan diabetic rabbits.
Ind. J. Pharmacol. 1997, V. 29, P. 162–167.
ДЕЯКІ НЕСПРИЯТЛИВІ ЕФЕКТИ,
ПОВ’ЯЗАНІ З ВИКОРИСТАННЯМ
БІОПРЕПАРАТУ З ЕКСТРАКТУ
НАСІННЯ Picralima nitida
ЯК АНТИДІАБЕТИЧНОГО АГЕНТА
О. А. Акінлойє1, Є. А. Балогун2,
Т. А. К. Омотанзе1, O. O. Аделейє1
1Відділ біохімії, Федеральний університет
сільського господарства, Aбеокутa, Нігерія
2Відділ ветеринарної патології, Федеральний
університет сільського господарства,
Aбеокутa, Нігерія
E-mail : oaakin@yahoo.com
За допомогою біохімічних і гістологічних
методів вивчено деякі несприятливі ефекти,
які можуть бути пов’язані з використан ням
біопрепарату з насіння P. nitida як гіпо глі-
кемічного засобу. Антирадикальну ак тивність
біопрепарату визначали за допомогою стабіль-
ного радикала 1,1-дифеніл-2-пікрилгідразилу.
Біохімічні дослідження включали визначення
активності аланін- і аспартатаміно-, гамма-
глутамілтрансферази, концентрації глюкози
крові, електролітів (натрій, калій і бікарбо-
нат), рівня пероксидного окиснення ліпідів.
Визначали також параметри крові (кількість
НЕКОТОРЫЕ НЕБЛАГОПРИЯТНЫЕ
ЭФФЕКТЫ, СВЯЗАННЫЕ
С ИСПОЛЬЗОВАНИЕМ БИОПРЕПАРАТА
ИЗ ЭКСТРАКТА СЕМЯН Picralima nitida
В КАЧЕСТВЕ АНТИДИАБЕТИЧЕСКОГО
АГЕНТА
О. А. Акинлойе1, Е. А. Балогун2,
Т. А. К. Омотанзе1, O. O. Аделейе1
1Отдел биохимии, Федеральный университет
сельского хозяйства, Aбеокута, Нигерия
2Отдел ветеринарной патологии,
Федеральный университет сельского
хозяйства, Aбеокутa, Нигерия
E-mail : oaakin@yahoo.com
С помощью биохимических и гистологиче-
ских методов изучены некоторые неблагопри-
ятные эффекты, которые могут быть связаны с
использованием биопрепарата из семян P. nitida
в качестве гипогликемического средства.
Антирадикальную активность биопрепара-
та определяли с помощью стабильного радика-
ла 1,1-дифенил-2-пикрилгидразила. Биохими-
ческие исследования включали определение
активности аланин- и аспартатамино-, гам-
маглутамилтрансферазы, концентрации глю-
козы крови, электролитов (натрий, калий и
Experimental articles
105
сумарних клітин крові, еритроцитів і лімфо-
цитів), функцій печінки і нирок, а також
ліпідний профіль. Гістопатологічні дослі джен-
ня печінки, нирок і підшлункової залози було
виконано за допомогою стандартного методу
фарбування гематоксилін-еозином.
За введення щурам Вістар максимальну
антиоксидантну активність порівняно з кон-
трольними розчинами (водний і водно-коко-
совий екстракти) мав метанольний екстракт
насіння (36,73%), що вказує на його висо-
кий ступінь нейтралізації вільних радикалів,
далі водний (19,36%) і водно-кокосовий
екстракти (4,09%). У всіх оброблених щурів
наприкінці експерименту спостерігали значне
зниження (Р < 0,05) концентрації глюкози кро-
ві (в діапазоні від 41,66% до 55,59%) та істотне
збільшення (Р < 0,05) маси тіла (в межах від
9,26% до 38,89%). Для всіх груп, які отримува-
ли екстракт насіння, характерним було збіль-
шення гематологічних показників (Р < 0,05),
а також достовірне (Р < 0,05) зниження пара-
метрів ліпідних профілів і активності ензимів
плазми. У всіх групах, які отримували екстр-
акт, спостерігали іонорегуляторні порушення,
зокрема гіперкаліє- і гіпернатріємію, однак
вони були значно меншими (Р < 0,05) напри-
кінці обробки. У всіх дослідних групах були іс-
тотно нижчі концентрація сечовини і бікарбо-
нату та рівень пероксидного окиснення ліпідів.
Гістопатологічні дослідження показали, що
екстракти не коригувати деякі спостережувані
патологічні стани тканин, індуковані діабетом.
Хоча, згідно з даними літератури, цукровий
діабет супроводжується різними гістологічни-
ми змінами в різних органах, одержані в цій
роботі результати морфологічного вивчення
тканини підшлункової залози в групах тварин,
які отримували і не отримували біо препарат,
свідчать про наявність ушкоджень різного сту-
пеня, однак їх тяжкість в осередках ураження
була більш виражена за введення біопрепарату
екстракту насіння. У зв’язку з цим одержані
дані, які дали змогу визначити гіпоглікеміч-
ну і антиоксидантну активність біопрепарату
з екстрактів насіння Picralima nitida як анти-
діабетичного засобу, дають підстави стверджу-
вати, що використовувати його для лікування
цього захворювання слід з обережністю.
Ключові слова: цукровий діабет, екстракт
насіння Picralima nitida, біохімічні,
гістологічні дослідження.
бикарбонат), уровня пероксидного окисления
липидов. Определяли также параметры крови
(число суммарных клеток крови, эритроци-
тов и лимфоцитов), функций печени и почек,
а также липидный профиль. Гистопатологи-
ческие исследования печени, почек и подже-
лудочной железы были выполнены с помощью
стандартного метода окрашивания гематокси-
лин-эозином.
При введении крысам Вистар максималь-
ной антиоксидантной активностью по сравне-
нию с контрольными растворами (водный и
водно-кокосовый экстракты) обладал метаноль-
ный экстракт семян (36,73%), что указывает на
его высокую степень нейтрализации свободных
радикалов; затем следовал водный (19,36%)
и водно-кокосовый экстракты (4,09%). У всех
обработанных крыс в конце эксперимента на-
блюдали значительное снижение (Р < 0,05)
концентрации глюкозы крови (в диапазоне от
41,66% до 55,59%) и существенное увеличе-
ние (Р < 0,05) массы тела (в пределах от 9,26%
до 38,89%). Для всех групп, получавших экс-
тракт семян, характерным было увеличение
гематологических показателей (Р < 0,05), а
также достоверное (Р < 0,05) снижение пара-
метров липидных профилей и активности эн-
зимов плазмы. Во всех группах, получавших
экстракт, наблюдали ионорегуляторные на-
рушения, в частности гиперкалие- и гиперна-
триемию, но они были значительно менее вы-
ражены (Р < 0,05) в конце обработки. Также во
всех опытных группах была существенно ниже
концентрация мочевины и бикарбоната, а так-
же уровень пероксидного окисления липидов.
Гистопатологические исследования показали,
что экстракты не корригировали некоторые на-
блюдаемые патологические состояния тканей,
индуцированные диабетом. Хотя, согласно дан-
ным литературы, сахарный диабет сопровожда-
ется различными гистологическими изменени-
ями в различных органах, полученные в этой
работе результаты морфологического изучения
ткани поджелудочной железы в группах живот-
ных, получавших и не получавших биопрепа-
рат, свидетельствуют о наличии повреждений
различной степени, однако степень их тяжести
в очагах поражения была более выражена при
введении биопрепарата из экстракта семян.
В связи с этим полученные данные, которые
позволили определить гипогликемическую и
антиоксидантную активность биопрепарата из
экстрактов семян Picralima nitida в качестве
антидиабетического средства, дают основание
утверждать, что использовать его для лечения
этого заболевания следует с осторожностью.
Ключевые слова: сахарный диабет, экстракт
семян Picralima nitida, биохимические и
гисто логические исследования.
... Not available in literature Alkaloids, polyphenols and terpenoids (Ogbonnia et al., 2008) Akuammicine, 10-deoxya-kuammine, akuammine, akuammidine, burnamine and picraline (Teugwa et al., 2013) (continued on next page) Acute toxicity: Oral administration of 600, 750, 1000, 1500 and 3000 mg/kg to mice did not exert any mortality Subacute toxicity: Toxic effect on the liver, kidneys and the lungs (Kouitcheu Mabeku et al., 2008). Pancreas degeneration in methanol seeds extract treated groups (Akinloye et al., 2014) Apocynaceae: Rauvolfia vomitoria Afzel. ...
Article
Ethnopharmacological relevance: The management of diabetes mellitus management in African communities, especially in Gabon, is not well established as more than 60% of population rely on traditional treatments as primary healthcare. The aim of this review was to collect and present the scientific evidence for the use of medicinal plants that are in currect by Gabonese traditional healers to manage diabetes or hyperglycaemia based here on the pharmacological and toxicological profiles of plants with anti-diabetic activity are presented in order to promote their therapeutic value, ensure a safer use by population and provide some bases for further study on high potential plants reviewed. Materials and methods: Ethnobotanical studies were sourced using databases such as Online Wiley library, Pubmed, Google Scholar, PROTA, books and unpublished data including PhD and Master thesis, African and Asian journals. Keywords including 'Diabetes', 'Gabon', 'Toxicity', 'Constituents', "hyperglycemia' were used. Results: A total of 69 plants currently used in Gabon with potential anti-diabetic activity have been identified in the literature, all of which have been used in in vivo or in vitro studies. Most of the plants have been studied in human or animal models for their ability to reduce blood glucose, stimulate insulin secretion or inhibit carbohydrates enzymes. Active substances have been identified in 12 out of 69 plants outlined in this review, these include Allium cepa and Tabernanthe iboga. Only eight plants have their active substances tested for anti-diabetic activity and are suitables for further investigation. Toxicological data is scarce and is dose-related to the functional parameters of major organs such as kidney and liver. Conclusion: An in-depth understanding on the pharmacology and toxicology of Gabonese anti-diabetic plants is lacking yetthere is a great scope for new treatments. With further research, the use of Gabonese anti-diabetic plants is important to ensure the safety of the diabetic patients in Gabon.
Article
Full-text available
L'infection à Trypanosoma brucei a conduit à un processus aigu et fatal chez les chiens communs nigérians en raison d'une anémie à évolution rapide. Les chiens infectés ont répondu par une réticulocytose accrue qui ne s'est pas prolongée avec la chronicité. En comparaison, la réponse à une anémie hémolytique artificiellement induite a été progressive, bien marquée et prolongée. L'anémie consécutive à l'infection à T. brucei chez le chien était soit normocytique et normochromique dans le cas d'une infection aiguë, soit microcytique et normochromique dans l'infection chronique. Quant à l'anémie induite artificiellement, elle était soit macrocytique et normochromique, soit normocytique et normochromique. Le potentiel érythropoïéique du plasma in vivo chez la souris était plus élevé chez les chiens infectés, sauf à la fin de la parasitémie. Cette anémie à T. brucei chez les chiens infectés est donc une réaction initiale mais devient un facteur de peu d'importance avec l'établissement de la chronicité.
Article
Full-text available
The hypoglycaemic and hypolipidaemic effects of the increasing dosages of A. cepa aqueous extracts on alloxan - induced diabetic Rattus novergicus for possible use in the management of diabetes mellitus was investigated. Diabetes mellitus was induced in 54 out of a total of 63 adult Rattus novergicus using 150mg/kg of alloxan monohydrate. Increasing dosages (200, 250 and 300mg/kg) of A. cepa aqueous extracts were given to the diabetic rats for six weeks while the control rats got either normal saline (1ml) or increasing dosages of glibenclamide (2.5, 3.8 and 5.0mg/kg) during the same period. Blood glucose level, total serum lipids and total serum cholesterol were assessed with routine methods. F-LSD was employed to test significant differences (P < 0.05) among treatment means. Increasing dosages of A. cepa aqueous extracts produced a dosedependent significant (P < 0.05) reductions in the blood glucose levels, total serum lipid and total serum cholesterol when compared with that of the control rats. The most effective percentage reduction in blood glucose level, total serum lipids and cholesterol were observed at 300mg/kg. From the experimental findings, it is possible to conclude that A. cepa studied exhibited promising hypoglycaemic and hypolipidaemic activity in alloxan-induced diabetic rats. It's hypoglycaemic and hypolipidaemic effects could represent a protective mechanism against the development of hyperglycaemia and hyperlipidaemia characteristic of diabetes mellitus.
Conference Paper
Type 2 diabetic patients have an increased risk of cardiovascular disease and, although many factors contribute to this risk, it is likely that diabetic dyslipidaemia plays an important role. Dyslipidaemia in Type 2 diabetic patients is characterized by low levels of I-IDL cholesterol and high triglyceride levels. In Type 2 diabetes, the total amount of LDL cholesterol is the same as in healthy people, but there are qualitative changes, e.g. a shift to smaller, denser LDL particles and an increased susceptibility to oxidation. Oxidized LDL may promote the development of atherosclerosis. It is possible to modify the major abnormalities of diabetic dyslipidaemia by combining lifestyle modifications (e.g. increased physical activity, cessation of smoking and weight reduction) with improved glycaemic control and hypolipidaemic drugs to reduce the burden of CVD within this high-risk population.
Article
Oral administration of 50% ethanolic extract of Cinnamomum tamala leaves significantly lowered (P < 0.01) the plasma glucose levels in normoglycaemic and streptozotocin hyperglycaemic rats. The extract also exhibited antihypercholesterolemic and antihypertriglyceridemic effects in streptozotocin-hyperglycaemic rats.
Article
Objective: The blood sugar lowering effect of P. nitida aqueous seed extract was investigated on normoglycaemic and hyperglycaemic rabbits. Materials and methods: The hypoglycaemic effect of P. nitida was evaluated using alloxan (80 mg/kg body weight, i.p) induced hyperglycaemic rabbits. The extracts potency was compared with standard drug, tolbutamide and distilled water. The LD50 was determined using mice. Results: A dose of 648 mg/kg body weight of extract caused maximum lowering of blood sugar levels in both normal and alloxanized rabbits. The mean fasting blood sugar in the normoglycaemic rabbit was reduced by 19.46% within 3 h, while in alloxanized rabbits blood sugar level was reduced by 75.5% within 6 h. The LD50 of the extract in mice was 1601.2 ± 60.5 mg/kg body weight when given i.p. Conclusion: P. nitida, though a crude drug, exhibited a faster onset of action and more persistent in hyperglycaemic situation than tolbutamide standard controls. This qualifies it to be used in ethnomedical diabetic management.