Journal of Acute Disease (2012)135-140 135

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Journal of Acute Disease

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Peperomia pellucida in diets modulates hyperglyceamia, oxidative stress

and dyslipidemia in diabetic rats
Rabiat U. Hamzah1*, Adebimpe A. Odetola2, Ochuko L. Erukainure3, Ademola A. Oyagbemi4
1
Department of Biochemistry, Federal University of Technology, Minna, Nigeria
2
Department of Biochemistry, College of Medicine, University of Ibadan, Ibadan, Nigeria
3
Food Technology Division, Federal Institute of Industrial Research, Lagos, Nigeria
4
Department of Veterinary Physiology, University of Ibadan, Ibadan, Nigeria

ARTICLE INFO ABSTRACT

Article history: Objective: To investigate the antidiabetic and antioxidant properties of Peperomia pellucida
Received 19 February 2012
(P. pellucida) in alloxan-induced diabetic rats. Methods: Beside mouse chow, two diets were
Received in revised form 30 March 2012
Accepted 7 April 2012
designed to contain 10%w/w and 20%w/w P. pellucida as supplements respectively. Diabetes
Available online 20 November 2012 was induced in groups of five male albino rats by a single intraperitoneal injection of alloxan.
Two groups of diabetic rats and normo rats were each fed one of these diets respectively, while
two other groups served as positive and negative controls respectively. A seventh group was
Keywords: fed pelletized mouse chow. Results: Diabetic rats on diets supplemented with 10%w/w and
Peperomia pellucid 20%w/w of P. pellucida for 28 d resulted in reduction of blood glucose level. The level of total
Antidiabetic serum cholesterol, triglycerides and LDL-cholesterol decreased significantly (P<0.05) with the
Antioxidant supplementation diets compared to the untreated diabetic rats. Also treatment with glibenclamide
and P. pellucida (10% and 20%w/w) led to increased activities of SOD, CAT and GSH respectively.
There was significant (P<0.05) reduction in the level of HDL-cholesterol, Catalase, SOD activities
and GSH concentration in diabetic untreated rats. The supplemented diets significantly (P<0.05)
reduced lipid peroxidation, which was elevated in untreated diabetic rats. Significant decrease
(P<0.05) in the activities of AST, ALT and ALP was also observed in rats fed P. pellucida
supplemented diets. Conclusions: The results from this study indicate that P. pellucida has an
antidiabetic and antioxidant properties in experimental diabetes mellitus and thus justifies the
acclaimed traditional antidiabetic use.

that diabetes is a condition within the body where the beta

1. Introduction cells of the islets of Langerhans in the pancreas do not
produce enough insulin and/or the insulin receptors are
Diabetes mellitus is a major public health problem in the not working properly. This results to an inadequate supply
developed as well as developing countries. It is ranked as of insulin and therefore elevated blood glucose level. The
the seventh cause of death in the world and third when symptoms of diabetes include increased blood glucose;
it’s fatal complications are taken into consideration[1]. It increased appetite and thirst, unexplained weight loss,
is a disease characterized by chronic hyperglycaemia and weakness, decreased blood pressure and blurred vision[4].
glucosuria produced by an absolute or relative insufficiency Diabetes is also associated with significant oxidative
of insulin. The ailment may result into the development of stress which has been reported to be a major contributory
further metabolic and anatomic disturbances among which factor to several diabetic complications[5].Reactive oxygen
is lipemia, hypercholesterolaemia, weight loss, ketosis, species (ROS) are important part of the defense mechanisms
arteriosclerosis, and gangrene, pathologic changes in the against infection, but its excessive generation has been
eye, neuropathy, renal disease and coma [2,3]. It is known implicated in the pathogenesis of vascular disease[6-8].
*Corresponding author: Rabiat U. Hamzah, Department of Biochemistry, Federal Diabetic patients have an increased incidence of vascular
University of Technology, Minna, Nigeria.
Tel: +2348035911608
disease and it has been shown that free radical activity is
E-mail: rabiune@yahoo.com;
136 Rabiat U. Hamzah et al./ Journal of Acute Disease (2012)135-140

elevated during diabetes[9,10]. Increased oxidative stress obtained from the faculty of Veterinary Medicine, University
has also been proposed to be one of the major causes of the of Ibadan, Ibadan, Nigeria were used for the study. They
hyperglycemia-induced trigger of diabetic complications. were fed on standard rat pellet diet (Ladokun Feeds, Nigeria)
Hyperglycemia stimulates ROS formation from a variety of and water was provided ad libitum. They were maintained
sources. These sources include oxidative phosphorylation, under standard laboratory conditions and were subjected
glucose autooxidation, NAD(P)H oxidase, lipooxygenase, to natural photoperiod of 12 h light: dark cycle. Group one
cytochrome P450 monooxygenases, and nitric oxide synthase. animals were administered with physiological saline at 10
Normal levels of antioxidant defense mechanism is not mL/kg body weight and served as control. Experimental
sufficient for the eradication of free radical induced injury, protocols complied with the “Principle of Laboratory Animal
therefore administrations of antioxidants from a natural Care” (NIH publication No 85-23) guidelines.
origin have a promising role to play. Several antioxidants
of plant material are experimentally confirmed and widely
2.4. Alloxan Induced diabetes
used as more effective agents against oxidative stress[11,12].
Peperomia pellucida (P. pellucida) (L.) HBK (Piperaceae) is
Diabetes was induced by a single (I.P) injection of 100 mg/kg
popularly known in Nigeria as shiny bush or riri and is used
of alloxan monohydrate. After 72 h of alloxan injection, the
locally for hypertension, diabetes and generally as tonic for
diabetic rats (glucose level > 250 mg/dL) were separated and
healthy well being. It is an herbaceous plant with succulent
alternate and ovate leaves, with terminal and axillary used for the study.
efflorescences, at the opposite side from leaves, developing
well in loose and humid soil by the tree shadows[13]. In folk 2.5. Preparation of reference drug
medicine, this species is employed on abscesses, furuncles,
and skin sores, as well eye inflammation (conjunctivitis). The reference drug, Glibenclamide (Clamide by hovid,
Literature data confirmed the antimicrobial and analgesic Malaysia) was purchased from Danax Pharmaceuticals, a
effects including other activities, such as anti-inflammatory local chemist in Ibadan, Nigeria. It was administered orally
effect[14,15]. Phytochemical studies revealed the presence of to the group on standard drug daily. The drug was dissolved
dill-apiol and pellucidin A, in P. pellucida[16]. The present freshly in normal saline and appropriate volumes were given
study aims at investigating the antidiabetic and antioxidant to the animals depending on their weight. The animals were
effect of P. pellucida in alloxan induced diabetic rats. given 600 毺g/kg body weight of the active ingredient.

2.6. Experimental design

2. Materials and methods
A total of 35 rats were used. The rats were randomly
2.1. Preparation of plant material distributed into seven groups of five rats each.
Group A - received water; standard feed and served as
F resh leaves of P. pellucida were collected from the Normal control (10 mL/kg bwt)
environs of the University of Ibadan, Ibadan, Nigeria. They Group B - Normal + 10%w/w of P. pellucida
were authencitated at the Herbarium, Botany Department, Group C - Normal + 20%w/w of P. pellucida
University of Ibadan, Ibadan, Nigeria. The leaves were air
Group D - Diabetic control
dried under laboratory conditions and grinded to powdering
Group E - Diabetic + 600 毺g/kg body weight glibenclamide
form. The fine powder was stored in airtight containers at
(standard drug)
room temperature until use. 100 g and 200 g of P. pellucida
Group F - Diabetic + 10%w/w P. pellucida
were compounded with 900 g and 800 g of standard rat
Group G - Diabetic + 20%w/w P. pellucida
feed (Ladokun feeds, Ibadan) to get 10% and 20% w/w of the
The rats were treated for four weeks after which they were
plant-supplemented diet respectively.
sacrificed by cervical decapitation. Plasma was collected
2.2. Chemicals after centrifugation of collected blood samples at 3 000 g
for 10 min in MSC bench centrifuge (Beckman and Hirsch,
Alloxan was obtained from were purchased from Sigma Burlington, 10, USA).
Chemical Co. (St. Louis, MO, USA). All other chemicals and The livers from animals were removed and rinsed in ice
drugs used in this experiment were of analytical grade and - cold isotonic, 1.15% KCl solution. The liver samples were
the purest quality available. homogenized in four times of ice - cold isotonic phosphate
buffer, pH 7.4 and centrifuged at 10 000 g for 15 min to
2.3. Animals obtain the post mitochondria fraction (PMF). Both plasma
and PMF aliquots were stored at -4 曟 until use.
Male albino rats of Wistar strain weighing about 150-200 g
 Rabiat U. Hamzah et al./ Journal of Acute Disease (2012)135-140
137

2.7. Biochemical analysis

Blood glucose was determined by the Glucose Oxidase

method described by NCCLS[17]. Total plasma cholesterol
was determined by Roeschlau et al[18], plasma triglycerides
by enzymatic colorimetric method using Randox Kit[19].
Plasma HDL was assayed by the method of Lopes-Virella et
al[20] plasma LDL[21] plasma alanine aminotransferase (ALT)
and aspartate aminotransferase activity was determined
by following the method of Reitman and Frankel[22],using
R andox kit, A lkaline P hosphatase ( ALP ) activity was
determined by the optimized DGKC method using Randox
kit. The supernatant obtained from the centrifuged liver
homogenate was used for the following biochemical assays:
superoxide dismutase (SOD), catalase, reduced glutathione
(GSH) and lipid peroxidation (LPO)[23-26].

2.8. Statistical analysis

A ll values are expressed as mean依 S . E . M . D ata was

analyzed by one-way analysis of variance (ANOVA) followed
by Newman-Keuls multiple comparison tests. Differences
Figure 1. Effect of P. pellucida on blood glucose level in normal
of means were considered significant at P<0 . 05 using and alloxan induced diabetic rats. Results are expressed as Mean依
Graph-Pad Prism software Prism version 4.00 for Windows, Standard deviation, n = 5.
G raph P ad S oftware, S an D iego C alifornia USA , “www. *P<0.05 between blood glucose level at week 0 and week 4 in each
graphpad.com.” group; aP<0.05 between blood glucose level at week 4 of each group
and end of normal control; bP<0.05 between diabetic control and
diabetic groups.

3.Results An increase in blood glucose was observed in diabetic

control rats compared to normal control while treatment with
The hypoglycemic effect of P. pellucida supplemented diet glibenclamide, P. pellucida at 10% w/w and 20% w/w diet
is shown in Figure 1. supplementation resulted in a 62%, 64% and 68% reduction

Table 1
Effect of P. pellucida on lipid profile in normal and alloxan induced diabetic rats.
Group No Treatment Total cholesterol (mg/dL) TG (mg/dL) HDL (mg/dL) LDL (mg/dL)
Group A Normal control 17.60依4.51 9.60依0.89 6.40依2.19 8.80依4.64
Group B Normal + 10%w/w P. pellucida 15.00依3.81 8.00依7.18 8.60依2.51 5.70依2.52b
Group C Normal + 20%w/w P. pellucida 14.40依5.60 7.20依3.70 9.80依3.49 4.36依2.65*
Group D Diabetic control 27.60依7.83* 21.00依3.67* 3.40依1.34* 20.52依7.15*
Group E Diabetic + 600 毺g/kg Gilben. 19.00依6.09** 12.40依5.86** 5.80依1.92** 10.52依3.52**
Group F Diabetic + 10% w/w P. pellucida 17.50依5.30** 10.75依6.50** 6.75依2.36** 9.30依5.67**
Group G Diabetic + 20% w/w P. pellucida 15.00依5.77** 8.75依5.69** 8.25依1.26**c 8.75依1.54**
Result are expressed as Mean依Standard deviation (n = 5). *P<0.05 All groups compared with Normal control; **P<0.05 All diabetic treated
compared with Diabetic control; bP<0.05 the two normal treated with P. pellucida compared; cP<0.05 Diabetic treated with P. pellucida
compared with Diabetic + Glibenclamide.

Table 2
Effect of P. pellucida on AST, ALT and ALP in alloxan-induced diabetic rats (unit/L).
Group No Treatment AST ALT ALP
Group A Normal control 16.20依7.05 8.60依6.91 10.80依4.32
Group B Normal + 10%w/w P. pellucida 11.20依5.26 6.40依3.78 8.80依2.95
Group C Normal + 20%w/w P. pellucida 17.80依9.23 9.20依5.54 13.40依3.78b
Group D Diabetic control 31.20依14.53* 18.20依3.27* 26.20依4.34*
Group E Diabetic + 600 毺g/kg Gilben. 22.80依10.18 13.00依5.24** 15.00依9.51**
Group F Diabetic + 10% w/w P. pellucida 18.25依9.09 11.25依5.06** 12.25依3.30**
Group G Diabetic + 20% w/w P. pellucida 17.50依8.50 10.00依6.78** 10.75依1.50**
138 Rabiat U. Hamzah et al./ Journal of Acute Disease (2012)135-140

Table 3
Effect of P. pellucida on lipid peroxidation(LPO), gutathione concentration(GSH), superoxide dismutase(SOD) and catalase activities(CAT).
Treatment group LPO (mmol/mg protein) GSH (毺g/mg protein) SOD (unit/mg protein) CAT (units/mg protein)
Normal control 2.03依0.72 4.98依0.69 1.42依0.19 1.56依0.58
Normal + 10% w/w P. pellucida 1.58依0.42 5.88依0.85 1.54依0.52 2.18依1.06
Normal + 20% w/w P. pellucida 2.82依1.15 6.24依0.86* 1.62依0.73 2.54依0.85*
Diabetic control 4.31依2.36* 1.76依0.55* 0.32依0.17* 0.58依0.37*
Diabetic + 600 毺g/kg Gilben. 3.34依2.67 2.58依0.57* ** 0.62依0.20* ** 0.86依0.24
Diabetic + 10% w/w P. pellucida 2.06依0.44** 3.00依0.57* ** 1.19依0.52** c 1.10依0.32**
Diabetic + 20% w/w P. pellucida 1.84依0.58** 4.05依0.52* ** ac 1.37依0.47** c 1.38依0.28** c

in blood glucose level respectively. utilization of glucose by tissues[9,28].

I t was observed that the levels of total cholesterol, Our findings indicate that P. pellucida has antidiabetic and
triglycerides and LDL - cholesterol except HDL-cholesterol antioxidant activities. A general increase was observed in
were significantly (P<0.05) higher in case of alloxan induced the level of blood glucose in the diabetic control rats. This
hyperglycemic animals (Diabetic control) when compared increase is significant when compared to the normal control,
with normal control (group 1) animals while the values of normal rats fed with different percentages (10% and 20% w/
the above mentioned plasma lipid parameters were near to w) of P. pellucida and the various diabetic rats treated with
normal in case of animals receiving glibenclamide and diet glibenclamide and P. pellucida. The hypoglycemic action
supplemented with 10% and 20% P. pellucida. However, was more pronounced with both 10% and 20% P. pellucida
HDL-cholesterol increased significantly in these groups supplementation than glibenclamide treated group. This
compared to the diabetic control (Table 1). may therefore imply that the P. pellucida preparation at
T able 2 shows the activities of AST , ALT and ALP of 10% and 20% w/w supplement are slightly more potent than
experimental rats. D iabetic rats showed significantly glibenclamide at 600 毺g/kg body weight dose. Though no
more activities of plasma AST, ALT and ALP respectively mechanism to the effect of P. pellucida on blood glucose
compared to normal control. T reatment with all plant has been proposed, it could be suggested that the plant
supplemented diet and glibenclamide significantly reduced might contain substances that mimic the action of insulin
the activity of AST, ALT and ALP in the diabetic control rats just like the sulphonylureas that promote insulin secretion
(P<0.05). by closure of K+-ATP channels, membrane depolarization
Table 3 show the effect of P.pellucida on lipid peroxidation, and stimulation of Ca2+ influx, an initial key step in insulin
GSH concentration and the activity of antioxidant enzymes, secretion[29,30].
SOD and CAT. A statistically significant (P<0.05) increase It is reported that the derangement of glucose, fat and
in lipid peroxidation was observed in diabetic control protein metabolism during diabetes, results into the
animals compared to normal control and these values were development of hyperlipidemia[31,32]. Diabetic rats were
brought down to almost normal by the plant in diabetic observed to have increased plasma lipids, which are
rats and below normal value in non-diabetic rats. The responsible for several cardiovascular disorders[33]. The
two supplement ( 10 % w/w and 20 % w/w ) reduced lipid higher lipid levels seen in diabetic rats may be due to
peroxidation by 52% and 57% respectively more effectively increased mobilization of free fatty acids from peripheral
than glibenclamide (23%). GSH concentration, SOD and depots and also due to lipolysis caused by hormones[34].
CAT activities were reduced significantly in diabetic Supplementation with both supplements of P. pellucida
rats compared to the control; however supplementation produced significant beneficial effects in the lipid profile in
with the 10%w/w, 20%w/w P. pellucida supplements and alloxan-induced diabetic rats, reducing triglycerides, total
glibenclamide increased the activities of these antioxidants. cholesterol, LDL, and increasing HDL, significantly. Thus,
it can be concluded from this findings that the levels of
total plasma cholesterol, triglycerides and LDL-cholesterol
4. Discussion which are actually raised in diabetes can be lowered with
P. pellucida supplementation; this antihyperlipidemic
Diabetes mellitus is one of the most common chronic effect could represent a protective mechanism against
diseases and is associated with hyperglycemia, the development of cardiovascular disorder such as
hyperlipidemia, increased oxidative damage, glucosuria and atherosclerosis in diabetic patients.
other complications such as obesity and hypertension[27]. The hepatic enzymes AST, ALT and ALP were used as
Alloxan, 毬- cytotoxin induces chemical diabetes in a wide biomarkers to check for early acute hepatic damage. The
variety of animal species by damaging the insulin producing activities of AST and ALT are cytosolic marker enzymes
pancreatic 毬- cell resulting in a decrease in endogenous reflecting hepatocellular necrosis as they are released
insulin release which paves the way for the decreased into the blood after cell membrane damage. Therefore,
 Rabiat U. Hamzah et al./ Journal of Acute Disease (2012)135-140
139

elevated activities of ALT and AST in the circulation serve superoxide radical O2•- by accelerating its conversion to
as indicators of hepatic damage. ALP acts as a marker for H2O2. Catalase is ubiquitous to most aerobic cells in animals
biliary function and cholestasis. In this study, all treatment and is especially concentrated in the liver and erythrocytes.
groups with experimental plant preparation effectively It is known to decompose hydrogen peroxide into water and
reduced plasma AST, ALT and ALP activities in diabetic rats oxygen as shown in the equation:
better than the standard drug, suggesting that the plants may 2H2O2 → 2H2O + O2 (1)
prevent hepatic injury associated with diabetes. The result showed that there was a significant reduction
Oxidative stress resulting from enhanced free radical in SOD and CAT activity in diabetic rats compared to
formation and/or defects in anti-oxidants defense causes normal control while all treated groups showed significant
severe tissue damage which may lead to a number of increase in activity with 20% w/w supplement showing
diseases like coronary artery disease, atherosclerosis, cancer greater increment compared to either group treated with
and diabetes. Increased oxidative stress in streptozotocin the drug and the 10% w/w supplement. This decrease in the
diabetic rats has been reported[35].This oxidative stress activity of SOD observed in diabetic rats is consistent with
is also implicated in the development of diabetic various reports already documented[40,41]. As proposed by
complications[36]. Increased oxidative stress as measured by Wohaieb and Godin[41], the reduction in SOD activity might
indices of lipid peroxidation and protein oxidation has been be due to the direct damaging effect of free radicals on the
shown to be elevated in both type1 and type 2 diabetes even enzyme. Therefore the activity of SOD, which is to remove
in patients without complications[37,38]. the superoxide radical O2- by accelerating its conversion
The results showed increased lipid peroxidation in the liver to H2O2 is inactivated. The ability of the plant preparation
of diabetic control group. This may be because the tissues to increase catalase activity may be due to the induction of
contain relatively high concentration of easily peroxidizable the enzyme. Catalase is haemoprotein and phytochemical
fatty acids. The increase in oxygen free radicals in diabetes screening of P. pellucida has also shown the presence of iron
could be primarily due to increase in blood glucose levels, (Fe)[42]. Iron is essential for the function of catalase.
which upon auto-oxidation generate free radicals and T he results of the present study show that diet
secondarily due to the effects of diabetogenic agent of supplemented P. pellucida have an antidiabetic and
alloxan [39]. In diabetes, hypoinsulinaemia increases the antioxidant property in alloxan induced diabetic rats.
activity of the enzyme, fatty acyl coenzyme, coenzyme A This effect may be due to the presence of tannin, saponin,
oxidase, which initiates 毬-oxidation of fatty acids resulting flavonoid and other constituents in the plant, which could
in lipid peroxidation[9]. Increased lipid peroxidation impairs act synergistically or independently in enhancing the
membrane functions by decreasing membrane fluidity, and activity of glycolytic and antioxidant enzymes. Therefore
changing the activity of membrane-bound enzymes[36]. Its the acclaimed traditional antidiabetic use of this plant is
products (lipid radicals and lipid peroxide) are harmful justified in this study.
to the cells in the body and are associated with various
disease condition, atherosclerosis and brain damage[36].
S upplementation with P. pellucida and glibenclamide Conflict of Interest
reduced the extent of lipid peroxidation in the diabetic
treated group. This indicates that P. pellucida supplement There was no conflict of interest among the authors.
may inhibit lipid peroxidation and thereby oxidative damage
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