Int J Biol Med Res.

2023 ;14(4):7665-7668
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Original article
Exploring the Protective Effects of Vitamin C and D against Phosphine-Induced
Kidney Damage
a
Akinpelu MORONKEJI, b2Frederick O AKINBO, cSolomon Matthias GAMDE*, aTemidayo Daniel ADENIYI, dAbiodun Abioye OYELEKE,
e
IDOWU-Aiye Faith
a
Department of Medical Laboratory Science, Faculty of Allied Health Sciences, University of Medical Sciences, Ondo State, Nigeria.
b
Department of Medical Laboratory Science, Faculty of Basic Medical Sciences, University of Benin, Edo State, Nigeria
c
Department of Medical Laboratory Science, Faculty of Allied Health Sciences, Bingham University Karu, Nasarawa State, Nigeria
d
Department of Anatomy, Ladoke Akintola University of Technology, Ogbomoso, Oyo State.
e
Department of Medical Laboratory Science, Achievers University Owo, Ondo State, Nigeria.

ARTICLE INFO ABSTRACT

Keywords:
Background: In Africa, Aluminium phosphide is commonly used for storage and transportation
Phosphide poisoning
of food grains. Unfortunately, the chemical build up in the food we eat is poisonous without a
Inflammation
Interstitial haemorrhage specific antidote. We speculate that the antioxidant effects of vitamins C and D could mitigate
Vitamins C and D. the long list of side effects. Aim of the Work: To determine the possible protective effect of
vitamins C and D against phosphine-induced kidney damage in Wistar rats Materials and
Method: This is an experimental laboratory-based study. Twenty Wistar rats were assigned
randomly into five groups (n=4). Group, I served as the normal control while Groups II-V were
exposed to 2.5 mg/kg phosphine tablet. Group II was the positive control without treatment
while Groups III-V were treated with vitamin C (100mg/kg/bw), vitamin D (10mg/kg/bw).
Group V was co-administered with vitamins C and D. All treatments lasted for 30 days. Animals
were euthanized and blood was collected via cardiac puncture for renal function parameter
while excised kidney was processed histologically by the paraffin wax method. Result: There
was a significant increase in the urea and creatinine levels in phosphine-induced animals,
indicating the kidney was injured. Creatinine is a more reliable marker for assessing kidney
function than urea. Moreover, a number of interstitial foci of haemorrhage and inflammatory
cells observed on the kidney confirmed the renal toxicity of phosphine. However, the co-
administration of vitamins C and D produced a dose-related improvements in the kidney
parenchyma. Conclusion: Aluminium phosphide is toxic to the kidney parenchyma. However,
co-administration of vitamins C and D ameliorates phosphine-induced kidney damage.
c
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1. Introduction
Nowadays, environmental pollution is a common health include oral administration of charcoal, coconut oil, and
problem confronting humans. Many pollutants interfere with our intravenous magnesium sulphate (4).
daily lives whether directly or indirectly, but the residual build up
Exposure to the toxic phosphine gas may reduce both humoral
of pesticides in the food we eat is one of the most toxic
and cellular immune functions which causes oxidative stress,
contaminants. According to World Health Organization (WHO),
depletion of the antioxidant levels, and nephrotoxicity (5). In our
millions of pesticide poisoning occur yearly accounting for over
previous study, we demonstrated and reported the mitigating
250,000 deaths (1). Aluminum phosphide (ALP) alone account for
effects of the co-administration of vitamins C and D in phosphine-
over 10.5% suicidal cases in 2019 (2). Unfortunately, there is no
induced liver damage (6). Moreover, Adikwu and Deo (7) reported
specific antidotes for phosphine poisoning consequently
that antioxidant supplementation especially vitamins C and
accounting for the high morbidity and mortality rates particularly
vitamin D ameliorate the stress associated with metal poisoning.
among the youths (3). Some of the palliative care for ALP poisoning
Similarly, in many experimental studies, vitamins C and D
diminishes reactive oxygen species and heightens the antioxidant
* Corresponding Author : Solomon Matthias Gamde defence system (8,9,10). Besides, vitamin D is essential for a wide
Department of Medical Laboratory Science,
range of non-classical functions and its deficiency is associated to a
Faculty of Basic Medical Sciences,
number of chronic disorders including diabetes, cardiovascular
University of Benin, Edo State, Nigeria
email; solomonmatthias85@gmail.com.
and kidney diseases (11).
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 Akinpelu MORONKEJI et al. /Int J Biol Med Res.14(4):7665-7668
7666

Some clinical studies on humans have associated positive health mean, while the student t-test was used to assess if there is any
outcomes with vitamins C and D ingestion (4), however their difference in the data obtained, p values < 0.05 was considered
significance are inexhaustible. From the perspective that the kidney statistically significant.
is susceptible to more than a few damage due to high volume of blood
RESULT
and toxins flowing through it and its ability to filter, there is paucity of
information on the histopathology of phosphine-induced kidney Biochemical assessment
damage and their possible antidote. It is against this background we
In this study, there was a significant increase in plasma urea and
determined the proctective effect of vitamins C and D on phosphine-
creatinine levels in phosphine-induced animals compared to the
induced kidney damage.
control group, indicating that the kidney was injured (Table 1).
MATERIALS AND METHODS
Table 1: Effect of Vitamin C and D on kidney function markers:
Ethical approval
This study was carried out in accordance with the code of ethics
for experiments involving the use of laboratory animals. The
protocol was approved by The Ethics and Research Committee, Ondo
State Ministry of Agriculture, Akure City, Ondo State, Nigeria as
referenced (MNR/V384/9).
Experimental animals
This is an experimental laboratory-based study. Animals were
obtained from the Animal Holding of the Department of Anatomical
Science, Achievers University Owo, Ondo State, Nigeria. The animals
were housed in ventilated cages at the Animal House and allowed Results were expressed as mean value ± standard deviation
access to a standard rodent diet and water ad libitum through out the (SD).*mean values were significantly different compared to the
experiment following the International Human-Animal Care Normal control at P≤ 0.05.#mean values were significantly different
Standard. compared with the positive control (Aluminum Phosphate). Group I:
Control, Group II: Aluminum Phosphate, Group III: Aluminum
Study Design Phosphate + Vitamin C, Group IV: Aluminum Phosphate + Vitamin D,
This is an experimental study. Twenty Wistar rats were assigned Group V: Aluminum Phosphate + Vitamin C and D.
randomly into five groups (n=4). Group, I served as the normal
control while Groups II-V were exposed to 2.5 mg/kg phosphine Figure 1: The effect of Vitamin C and D on electrolytes and
tablet. Group II served as the positive control without treatment creatinine values. *mean values were significantly different
while Groups III-V were treated with vitamin C (100mg/kg/bw) and compared to the phosphine group at P≤ 0.05.
vitamin D (10mg/kg/bw). Group V was co-administered with
vitamins C and D. All treatments lasted for 30 days and the animals
were euthanized for sample collection.
Sample collection
Blood was collected via cardiac puncture into a lithium
heparinized anticoagulant bottles and was centrifuged at 3000 rpm
for 15 mins to obtain the plasma. The plasma obtained was used for
biochemical analysis while the excised kidney was fixed in 10%
neutral buffered formalin for histological investigation.
Biochemical analysis
Plama urea (U) and creatinine (Cr) were analysed using
diagnostic kits from Randox laboratory, United Kingdom while
plasma electrolytes; potassium (K+), sodium (Na+), chloride (Cl-),
Figure 2: Effect of vitamin C and D on potassium and urea levels.
and bicarbonate (HCO3) were estimated using “ARCHITECT c4000”
*mean values were significantly different compared to the
Aboot automated chemistry analyser.
phosphine group at P≤ 0.05.
Histopathological analysis
Representative organs were processed histologically as
described by Avwioro (12). Briefly, the grossed tissue was
dehydrated in three changes of alcohol, xylene, and molten paraffin
wax. Sections was cut at 4 microns on a rotary microtome (Leica
RM2125 RTS, Leica Biosystems, Buffalo Grove, United States of
America) and stained in hematoxylin-eosin (H & E).
Statistical analysis
Data were evaluated by one-way analysis of variance (ANOVA)
using statistical package for social science (SPSS) Version 25 (SPSS,
Cary, NC, USA) and expressed as the mean standard error of the
 Akinpelu MORONKEJI et al. /Int J Biol Med Res.14(4):7665-7668
Histopathology assessment
There was inflammatory cells and foci of interstitial
haemorrhage on the kidney parenchyma. However, co-
administration of vitamins C and D produced a dose-related
improvements in the kidney (Figures 3-5).
Figure 3 Normal kidney section (I): shows renal corpuscle
(white arrow), renal tubule (black arrow), and proximal tubule
(blue arrow). Phosphine induced degenerated kidney (II)
exhibited severe inflammation (red arrow). (H&E. Mag. X 400).
Figure 4 Phosphine induced degenerated kidney (I) treated
with Vitamin C exhibited severe inflammation (red arrow)
while animals treated with Vitamin D (IV) exhibited extravasion
of blood (red arrow). (H&E. Mag. X 400).
Figure 5 Phosphine induced degenerated kidney (V) treated
with Vitamin C and D shows mild interstitial haemorrhage (red
arrow) with improved histological features compared to
normal control (I). (H&E. Mag. X 400).
7667
DISCUSSION
The protective effect of antioxidants against the many
deleterious effects of ALP have been reported (13). We determined
the protective effects of vitamins C and D against phosphine-induced
kidney damage. In this experimental study, ALP caused significant
changes in both the biochemical and anatomic structures of the
kidney. There was a significant decrease in sodium (Na2+) levels in
the phosphine-induced group whereas vitamin supplementation
restored the Na2+ values with the most significant improvement
detected in the group co-administered vitamins C and D. There was
also significant restoration of potassium, chloride, and bicarbonate
values to normalcy in the group. Previous studies has suggested that
electrolytes abnormalities, such as high or low sodium and
potassium levels may be present in phosphine poisoning (14,15,16).
According to Mokhtari et al., (11), vitamin D improves the
antioxidant defence system such as glutathione, glutathione
peroxidase, and superoxide dismutase. Normal redox homeostasis
would break down when their intracellular scavenging capacity is
abridged. In some cases of metabolic stress, the levels of intracellular
oxidants may rise with toxic effects indiscriminately damaging
proteins, lipids, and DNA and activating specific signaling pathways
that is linked to the onset of a number of diseases (4). In addition,
some studies have also indicated the significance of vitamin C in
reducing the oxidative effect of phosphine (17,18). Creatinine is a
reliable marker than urea to assess kidney function (19), a
significant increase in both creatinine and urea in the present study
shows that the animals suffered nephrotoxic changes. The
alleviating effect of vitamins when administered individually, as well
as the co-administration of vitamins C and D was apparent in this
study with a significant reduction in urea and creatinine values.
Moreover, free radicals interacts with normal cellular components
leading to tissue-breakdown (14,20). The histopathological changes
in the present study is similar to previous reports (21,22) that
documented interstitial haemorrhage in the kidney of experimental
animals.
A number of studies have reported the role of vitamin D
supplementation in oxidative stress (23,24,25,26,27). The present
study revealed for the first time that vitamins C and D
supplementation improved the kidney function as well as histologic
features in phosphine induced damaged in tandem with previous
studies which established phosphine-induced kidney damage
(28,29).
CONCLUSION
In this study, ALP significantly increased the plasma urea and
creatinine levels, indicating that the kidney was injured. A number
of inflammatory cells and interstitial haemorrhage proved the
kidney was injured. However, co-administration of vitamins C and D
produced a dose-related improvements in the kidney.
Declarations:
Ethics Approval: The protocol was approved by The Ethics and
Research Committee, Ondo State Ministry of Agriculture, Akure City,
Ondo State, Nigeria as referenced (MNR/V384/9).
Consent for publication: Not applicable
Availability of data and material: Data are available from the
corresponding author upon request.
 Akinpelu MORONKEJI et al. /Int J Biol Med Res.14(4):7665-7668
7668
Competing interests: Nothing to declare
Funding: None
Acknowledgments: The authors acknowledge the Animal Ethics
Committee.
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