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Vol. 5(1), 14-21, January (2016) Int. Res. J. Biological Sci.

Attenuation effect of Moringa oleifera leaves powder on Blood Biochemical

disturbance induced in lead-exposed rats
Aïssi K.Alain1*, Casimir D. Akpovi1, Julien Sègbo1, MaximinSenou1, Eugénie Anago1, Théodora A. Ahoyo1, Jean R. Klotoé1,
Patrick A. Edorh2 and Frédéric Loko1
1
Research Laboratory in Applied Biology, Department of Human Bioengineering, Polytechnic School of Abomey-Calavi, University of Abomey-
Calavi, BENIN
2
Research Laboratory in Biochemistry and Environmental Toxicology, Department of Biochemistry and Cellular Biology, University of Abomey-
Calavi, BENIN
[email protected]
Available online at: www.isca.in, www.isca.me
Received 23rd November 2015, revised 24th December 2015, accepted 4th January 2016
Abstract
The nutritional and phytopharmaceutical composition of Moringa oleifera leaves suggest that this vegetable can prevent
toxic effects induced by xenobiotics .The current study aims to assess the prophylaxic capacity of Moringa oleifera leaves
powder (MoLP) on biochemical disturbances in lead-exposed rats. Twenty-eight (28) Wistar rats were equally divided into
four groups receiving respectively distilled water (Control), MoLP, lead acetate and both lead acetate + MoLP. The
treatment was administrated orally for 28 days. Blood samples were collected from animals at day zero (D0) and D28 for
biochemical assays. Results showed that lead treatment decreased significantly the levels of total proteinemia (p<0.01),
albuminemia (p<0.05), serum iron (p<0.01) and calcemia (p<0.01) compared to control group. MoLP addition to lead
treatment produced opposite effect on these parameter levels and reversed the impact of lead treatment. The levels of blood
urea (p<0.001), creatinine (p<0.01) and uric acid (p<0.01) increase significantly in lead-treated animals compared to
controls. MoLP treatment induced no significant difference in the levels of blood urea, creatinine and uric acid. However, it
suppresses the effect of lead treatment. On the whole, these results showed that MoLP administration to rats reverses lead
deleterious effects on many circulating biochemical parameters. The results highlight MoLP importance in the attenuation of
some biochemical disturbances associated with chronic lead absorption and strengthen arguments in favour of the use of
MoLP as food supplement.

Keywords: Lead toxicity, Moringa oleifera leaves powder, Biochemical disturbances, Blood, Rat.

Introduction M. oleifera leaf is the most commonly used part of the plant
because it’s easier accessibility for people8. Therefore, it is
Moringa oleifera is a vegetable species to Moringaceae family important to ensure the accuracy of various indications of MoLP
widespread in tropical and subtropical regions, particularly in which is increasingly sold as food supplement in West Africa9.
sub-Saharan Africa1. It is increasingly promoted worldwide for The current study aims at evaluate the prophylactic capacity of
its impressive range of medicinal use2 and high nutritional MoLP against biochemical disturbances induced by lead
value3. Several authors report the use of its leaves, roots, seeds, exposure.
barks, fruits, flowers and immature pods in the prevention or
treatment of many diseases such as diabetes, hypertension, Materials and Methods
inflammation, bacterial infections, fungal infestations, immune
deficiencies, heart disease, cancer, epilepsy, gastric ulcer, Chemical and vegetal materials: Lead acetate solution at 10%
urinary disorders, hyperlipidemia, and liver disease2. (Manufactured by HACH Company) was acquired at the
National Laboratory for Quality Control of Water and Food in
M. oleifera leaves hydro-alcoholic extract can be used to Ministry of Health. It was diluted in distilled water for a final
prevent adverse toxic effects of some antipyretic4 and ant concentration of 10 mg/ml.
tubercular drugs5. A detoxifying activity of the same hydro-
alcoholic extract was highlighted in lead-exposed rats. Indeed, Mature and fresh leaves of M. oleifera were harvested at Come,
the seed powder of this plant was used to reduce organic 60 km from Cotonou in Benin. The plant identity was
damages induced by arsenic6 and lead7. However, there are few authenticated by a botanist from University of Abomey-Calavi,
studies on the antitoxic potential of M. oleifera leaves powder Benin. After detaching, the leaves were washed following the
(MoLP), commonly used. procedure described by Sauveur and Broin10. Then, they were
dried away from direct sunlight and dust for fourteen days

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before to be milled. The obtained powder was recovered in a France) on a Mindray BS 200 auto-analyzer.
clean box and kept away from humidity. It was dissolved in
distilled water one day before each use for the rats. Statistical data processing: Data were entered in Excel 2010
then exported in SPSS 16.0 software. The mean and standard
Animal and experimental design: The study was done in deviation were calculated for each measured parameter. Values
University of Abomey-Calavi (Benin). Twenty-eight (28) were checked for homogeneity of variances using the Levene's
Wistar rats aged 10-12 weeks and weighing between 145-151 g test. A one-way analysis of variance followed by the post hoc
were equally divided into four groups. They were placed in Bonferroni’s multiple comparisons test was carried out for
identified cages inside a room where the light was alternated comparing mean levels and detect specific significances
with the darkness per cycle of 12 hours. All animals had free differences between groups. Differences were considered as
access to water and standard rodent diet which was renewed significant when p<0.05.
every morning.
Results and Discussion
The first group (Control) received 0.5 ml of distilled water. The
second group (MoLP) received 500 mg/kg body weight (bw) of Results: Total proteinemia: From D0 to D28, total serum
MoLP. The third group (Lead) received 10 mg/kg bw of lead protein level decreased significantly (p<0.01) by 29.3 % in the
acetate. The last group (Lead + MoLP) received both lead lead-exposed group (figure-1) compared to control. Rats
acetate (10 mg/kg bw) and MoLP (500 mg/kg bw). The treatment with MoLP or the combination of lead and MoLP
products were daily administered by force-feeding for 28 days. showed no significant variation in total protein level compared
Different probe was used for each kind of solution in order to to controls. MoLP has allowed keeping 32.2 % of the serum
avoid interferences. protein level that would be lost under the lead action (p<0.01).

Blood sampling and biochemical analysis: At the beginning Albuminemia: Albuminemia decreased significantly (p<0.01)
(D0) and end of the exposition period (D28), animals were by 29.3 % in lead-treated group (figure-2). MoLP addition to
anesthetized by exposure to chloroform vapour. Blood samples lead treatment has reduced significantly (p<0.01) by 34.3 % the
were collected by venipuncture in the retro-orbital plexus with a risk of decreasing in serum albumin level.
heparinised capillary pipette following the method described by
Tehoua et al.11. All specimens in collection tubes without Blood Urea: The level of uremia increased significantly
anticoagulant (Vacutainer system; Becton Dickinson) was (p<0.001) by 127.8% in the lead treated group (figure-3). No
labelled and centrifuged within 2 hours at 2500 rpm/min for 10 significant variation of uremia was noted in the other groups.
minutes. Total protein, albumin, urea, creatinine, iron, calcium, The supplementation with MoLP has reduced significantly
glucose, uric acid, total cholesterol and triglycerides were (p<0.01) by 112 % the risk of increasing in blood urea level.
measured by using Elitech reagents (ELITech Group, Maizy,

**p<0.01; Values with the same letter are not significantly different
Figure-1
Total proteinemia level in rats exposed to lead acetate and/or Moringa oleifera leaves powder compared to controls

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*p<0.05; Values with the same letter are not significantly different
Figure-2
Albuminemia level in rats exposed to lead acetate and/or Moringa oleifera leaves powder compared to controls

***p<0.001; Values with the same letter are not significantly different
Figure-3
Blood urea level in rats exposed to lead acetate and/or Moringa oleifera leaves powder compared to controls

Creatininemia: Lead treatment induced a significant increase supplementation with MoLP has reduced significantly (p<0.01)
(p<0.01) in creatinine level by 56.2 % (figure-4). MoLP by 43.1 % the risk of hyperuricemia.
addition to lead treatment has allowed preventingsignificantly
(p<0.01), 34.3 % the risk of increasing in creatininemia level. Serum iron: Serum iron level decreased significantly (p<0.01)
by 58.3 % in the lead-exposed group (figure-6). Rats treatment
Uricemia: The level of uric acid increased significantly with MoLP or the combination of lead and MoLP showed no
(p<0.05) by 73.1 % in the lead treated group and by 30 % in the significant variation in iron level compared to with the controls.
similar group which received MoLP in addition (figure-5). The MoLP has allowed preserving 52.7 % of the serum iron level
difference in those increased level indicate that the that would be lost under the lead action (p<0.01).

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**p<0.01; Values with the same letter are not significantly different
Figure-4
Creatininemia level in rats exposed to lead acetate and/or Moringa oleifera leaves powder compared to controls

**p<0.01; *p<0.05; Values with the same letter are not significantly different
Figure-5
Blood uric acid proteinemia level in rats exposed to lead acetate and/or Moringa oleifera leaves powder compared to
controls

Serum calcium: Serum calcium level decreased significantly Cholesterolemia: No significant variation in the serum
(p<0.01) by 21.8 % in lead-treated group (figure-7). MoLP cholesterol level was noted in the experimental groups
addition to lead treatment has reduced significantly (p<0.01) by compared to the control (figure-9).
16.8 % the risk of calcemia decreasing.
Triglyceridemia: The level of triglycerides did not vary
Blood glucose level: Blood glucose level showed no significant significantly in any of the treated groups compared to the
variation in rat group receiving lead alone, MoLP alone or control (figure-10).
association of both lead and MoLP when compared to control
(figure-8).

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**p<0.01; Values with the same letter are not significantly different
Figure-6
Serum iron level in rats exposed to lead acetate and/or Moringa oleifera leaves powder compared to controls

**p<0.01; Values with the same letter are not significantly different
Figure-7
Serum calcium level in rats exposed to lead acetate and/or Moringa oleifera leaves powder compared to controls

Discussion: This study showed that the administration of 10 nephrons, the structural and functional unit of the kidneys13.
mg/kg bw of lead acetate to Wistar rats induce a decrease in This justifies the observed hypercreatinemia which is a specific
serum protein and albuminemia levels. These findings are in markers showing the decrease in kidneys capacity to purify
agreement with some published results12. Who reported blood and excrete urine. The increase of blood urea and
hypoproteinemia in rats exposed to lead acetate during a week? creatinine in lead intoxicated rats is due to a nephropathy
Hypoproteinemia in lead-exposed rats is followed by a blood characterized by glomerular and/or tubular damages14. These
urea increase. This can be explained by proteins catabolism biochemical disturbances were correlated with the
increasing12 from which amino acids are converted into hyperuricemia observed in the lead-exposed rats. It was reported
ammonia leading to urea. Lead affects the excretion function of that lead can induce oxidant/antioxidant system imbalance15.

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Lead induced tubular reabsorption of uric acid14 which is an that can alter the metabolism16. The serum iron is rarely free16. It
oxidative stress biomarker16. The observed decreasing serum appears in blood only under pathological conditions after its
iron and calcium level may be correlated to this oxidative releasing from transport proteins because of the reactive oxygen
condition. The lead competes with these two metal ions, by species.
expelling them from the biding sites of proteins and other
transport macromolecules. Iron and calcium deficiency may In this study, glycemia has not been affected by lead
facilitate the rapid absorption of lead that may in return intoxication. Similar results have been reported19, but some
accentuate their deficiency17. The released iron because of the authors have found hypoglycaemia in lead-exposed rats20 while
lead, contributes to amplify the oxidative stress18 by catalysing others12 reported hyperglycaemia in rats treated with the same dose
the synthesis of free radicals and other pro-oxidative molecules of lead.

Values with the same letter are not significantly different

Figure-8
Blood glucose level in rats exposed to lead acetate and/or Moringa oleifera leaves powder compared to controls

Values with the same letter are not significantly different

Figure-9
Total cholesterolemia level in rats exposed to lead acetate and/or Moringa oleifera leaves powder compared to controls

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Values with the same letter are not significantly different

Figure-10
Triglyceridemia level in rats exposed to lead acetate and/or Moringa oleifera leaves powder compared to controls

Cholesterol and triglyceride are the two major blood lipids. The detoxification property of Moringa oleifera is also due to its
Lead intoxication didn’t affect the cholesterol level. This result high concentration in methionine and cysteine, which are amino
is in accordance with that of some authors21, but it contrasted acids with sulfhydryl group that improve heavy metals
with those of others22 who reported increased circulating free chelation26.
cholesterol in rats exposed to lead acetate during one month.
Blood triglyceride levels did not vary significantly. These Conclusion
results also contrasted with those of other authors21 who showed
that administration of 10 mg/kg bw decrease triglyceride levels The results of study highlight that Moringa oleifera leaves
in rats. powder has antitoxic properties. It has prevented some lead
deleterious effects on many circulating biochemical parameters
The administration of Moringa oleifera leaves powder in lead- in rats. These arguments are in favour of the use of Moringa
exposed rats has preserved them against protein, iron and oleifera leaves powder as food supplement especially in chronic
calcium deficiency. This observation can be explained by the lead-exposed animals.
richness of Moringa oleifera leaves in proteins, iron and
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