Effect of L-Glutamic acid on histology and functions of Liver and kidney of Rats and Protective Role of Zingibar Officionale

Abstract

Background: Monosodium glutamate (MSG) or L-Glutamic acid is the sodium salt of
glutamic acid is toxic to human and experimental animals. Liver and kidney may be
susceptible to injury resulting from toxic substances. Ginger has immuno-modulatory,
antitumorigenic, anti-inflammatory, anti-apoptotic, antihyperglycemic and anti-lipidemic
actions.
Objective: This study has been carried out to evaluate the protective effects of aqueous
extract of Zingiber Officinale in decreasing and ameliorating effects of Monosodium
glutamate induced alteration in hepatic and renal tissues and their functions of rats.
Patients and Methods: Twenty four adult rats were divided into three equal groups and
maintained under standard laboratory conditions. The ginger extract and the MSG were
given orally once daily for 21 days. Group I Control: distilled water Group II: received
MSG (4.0g/kg B.W) Group III: received 4.0g/kg B.W of MSG +100mg/Kg of ginger.
After the last dose blood was collected.
Results: Monosodium Glutamate /L-Glutamate caused loss of normal histological
architecture of liver and kidneys of rats of Group II with significant changes in biochemical
parameters compared with control groups (P < 0.0001). Zingibar Officinale /Ginger is an
excellent antioxidant substance that ameliorated and prevented the toxic effect of MSG in a
group III treated with combination of MSG and Ginger histologically and functionally.
Conclusion: Zingibar Officinale (ginger) caused decrease the hepatotoxic and nephrotoxic
effects resulted from oxidative damage induced by MSG because of their antioxidant
effects.

Full text

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Diyala Journal of Medicine Volume 11, Issue 2, December 2016
DJM
Diyala Journal of Medicine
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V
Table of Contents
No.
Paper Title
Page
1
Prevalence and Association of Atypical Cervical Epithelial Cell
Abnormalities with Different Sociodemographic Factors among Women
Attending the Maternity Teaching Hospital
in Erbil, Iraq.
Runak Ali Ismail , Samira Salim Shekh Mohammad and Zahida Haris Saeed
1-8
2
Role of Cyclin D1 in Patients with Chronic Hepatitis C Dina Abas karbool and Areej Atiyah Hussein
9-16
3
Distribution of Antithyroid Peroxidase Antibody in Patients with
Clinically Suspected Thyroid Dysfunction Mazin A Shakir , Sawsan M Abdil Majeed and Waleed AT Al Ani
17-24
4
Assessment of Neopterin and Interleukin-1 Beta Serum Levels in Burn
Patient Ruqaya M. Al-Barzinji and Amer A. Khaleel
25-33
5
Evaluation of the Effect of Varicocelectomy on Semen Parameters and
Fertility Saad Muhmood Hussain Arraki
34-43
6
Bactericidal activity of 5 kinds of disinfectant on Vibrio cholera isolated
from drinking water in Baqupa city Khadija Shabban Al-mizury and Amel Mutafa kamil
44-50
7
Effect of L-Glutamic acid on histology and functions of Liver and kidney
of Rats and Protective Role of Zingibar Officionale Snoor Jalal Mustafa,Gulala Ibrahim Qader and
ShlerAkram Faqe Mahmood
51-59
8
Histopathological findings in hysteroctomized sample of women with
abnormal uterine bleeding attending Al- Batool Maternity Teaching
Hospital. Baquba /Iraq Sarah Walied Rahiem , Sawsan Talib Salman and Walied Rahiem Mahmod
60-65
9
Ultraviolet A Is AnEfficient Therapy For Plane Warts
Alaa Aldeen Khorsheed Murad, Attaa A. Hajji and Mohammed Hussein Alqaisy
66-72
10
Bioeffects of 1.5TStatic magnetic field on the DNA strand of human
leukocytes in vitroduring MRI scan Hanaa N. Abdullah, Haydar A. Al-shimmari, Mudhafer B. Mahdi and Muhammed M. Radhi
73-77

Effect of L-Glutamic acid on histology and functions of Liver and kidney Snoor Jalal Mustafa
of Rats and Protective Role of Zingibar Officionale
Diyala Journal of Medicine 51 Vol.11, Issue2,December2016
Effect of L-Glutamic acid on histology and functions of Liver and
kidney of Rats and Protective Role of Zingibar Officionale
Snoor Jalal Mustafa (PhD) 1,Gulala Ibrahim Qader (Msc)2and
ShlerAkram Faqe Mahmood (Msc)3
Abstract
Background: Monosodium glutamate (MSG) or L-Glutamic acid is the sodium salt of
glutamic acid is toxic to human and experimental animals. Liver and kidney may be
susceptible to injury resulting from toxic substances. Ginger has immuno-modulatory,
antitumorigenic, anti-inflammatory, anti-apoptotic, antihyperglycemic and anti-lipidemic
actions.
Objective: This study has been carried out to evaluate the protective effects of aqueous
extract of Zingiber Officinale in decreasing and ameliorating effects of Monosodium
glutamate induced alteration in hepatic and renal tissues and their functions of rats.
Patients and Methods: Twenty four adult rats were divided into three equal groups and
maintained under standard laboratory conditions. The ginger extract and the MSG were
given orally once daily for 21 days. Group I Control: distilled water Group II: received
MSG (4.0g/kg B.W) Group III: received 4.0g/kg B.W of MSG +100mg/Kg of ginger.
After the last dose blood was collected.
Results: Monosodium Glutamate /L-Glutamate caused loss of normal histological
architecture of liver and kidneys of rats of Group II with significant changes in biochemical
parameters compared with control groups (P < 0.0001). Zingibar Officinale /Ginger is an
excellent antioxidant substance that ameliorated and prevented the toxic effect of MSG in a
group III treated with combination of MSG and Ginger histologically and functionally.
Conclusion: Zingibar Officinale (ginger) caused decrease the hepatotoxic and nephrotoxic
effects resulted from oxidative damage induced by MSG because of their antioxidant
effects.
Key words: Monosodium glutamate, ginger, antioxidant, liver, kidney
Corresponding Author:snoorm88@gmail.com
Received: 31thMay 2016
Accepted: 24thJuly 2016
1Department of Anatomy- School of Medicine-Faculty of Medical Sciences- University of
Sulaimani-Sulaimani- Iraq.
2 Department of Pharmacology- School of Medicine-Faculty of Medical Sciences- University
of Sulaimani - Sulaimani - Iraq.
3 Department of Microbiology - School of Medicine-Faculty of Medical Sciences- University
of Sulaimani - Sulaimani - Iraq.
Introduction
Monosodium Glutamate (MSG) or
L-Glutamic acid known as AJI-NOMOTO
is the sodium salt of glutamic acid [1].
Glutamate is one of the most common
amino acids found in nature and is the main
component of many proteins and peptides

Effect of L-Glutamic acid on histology and functions of Liver and kidney Snoor Jalal Mustafa
of Rats and Protective Role of Zingibar Officionale
Diyala Journal of Medicine 52 Vol.11, Issue2,December2016
of most tissues, it is also produced in the
body and plays an essential role in human
metabolism [2,3]. Now MSG can be
produced commercially by bacterial
fermentation [4]. Despite its taste
stimulation and improved appetite
enhancement, reports indicate that MSG is
toxic to human and experimental animals
[5]. The hepatocytes have metabolic
functions that deal with very essential
processes such as detoxification,
deamination, transamination, removal of
ammonia in the form of urea, biosynthesis
and release of the non-essential amino acids
and plasma proteins, gluconeogenesis,
storage of glycogen, conversion of
carbohydrates and proteins into lipids,
synthesis of lipoproteins, phospholipids and
cholesterol, oxidation of fatty acids, storage
of iron in the form of ferritin as well as
storage of vitamins A, D and B12. Several
functional tests have been formulated to
explore hepatic status [6,7,8,9,10]. Several
enzymes have been determined to explore
hepatic status such as alanine amino
transferase (ALT) and aspartate amino
transferase (AST) and because the liver is
involved in the performance of these varied
functions, it may be susceptible to injury
resulting from toxic substances [11].
The Kidneysarenormally involved with
the removal of toxic metabolites and waste
products from the blood and regulation of
the amount of fluid and electrolytes balance
in the body. To test functions of the kidneys
routine urinalysis is used to measure serum
urea, creatinine, sodium, potassium and
bicarbonate [7,12]. However several studies
in animals have shown that MSG is toxic to
various organs such as liver, brain, thymus,
and kidneys [1,2].
ZingebarOffecionale (Ginger) is a strong
anti-oxidant substance prevents generation
of free radicals. It is considered a safe
herbal medicine with only few and non-
significant side effects (13). The main
pharmacological actions of ginger
include immuno-modulatory,
antitumorigenic, anti-inflammatory, anti-
apoptotic,
antihyperglycemic, anti-lipidemic and anti-
emetic actions.
This study has been carried out to evaluate
the protective effects of aqueous extract of
ZingiberOfficionale on Monosodium
glutamate induced
histomorphometricalteration in hepatic and
renal tissues and their functions in rats.
Patients and Methods
Twenty four adults male rats weighing
between 180-210g were maintained in a well-
ventilated animal house under standard
condition of humidity, temperature and a
constant light: dark lighting schedule, they
were allowed to acclimatize for one week
prior to the start of the experiment. The
animals were fed with pelletized food and
water. They were obtained from the animal
house of veterinary college, University of
Sulaimani. The animals were housed in clear
polypropylene cages lined with wood chip
beddings. The health and reproductive status
of the animals were assessed and only
healthy animals were selected for the
experiment.
Statistical analysis
Synthetic monosodium glutamate (MSG)
of 98% purity was obtained from Aldrich
chemistry, France, for use in the study. A
stock solution was prepared by dissolving
(1.0g) MSG granule in (1.0) ml distilled
water. From this and based on the animals
weight, 4.0g/kg B.W dosage were
administered to the animals in group (II and
III) using gavage tube [14].
The aqueous extracts of Zingiber
Officinale (Ginger) was prepared by
dissolving 1 g of dried powderof MSG in
50ml of distilled water and the final extract
concentration obtained was 20mg/ml. Extract

Effect of L-Glutamic acid on histology and functions of Liver and kidney Snoor Jalal Mustafa
of Rats and Protective Role of Zingibar Officionale
Diyala Journal of Medicine 53 Vol.11, Issue2,December2016
was stored in air- tight container and
refrigerated throughout the experiment(15).
Experimental design:The twenty four rats
were divided into three groups of eight
animals per each.The ginger extract and the
MSG were given orally once a day for 21
days. They were administered as follows:
Control Group I: received distilled water
Group II: received MSG (4.0g/kg B.W)
Group III: received MSG 4.0 g/Kg
BW and ginger 100g/kg .
Blood was collected; serum was prepared by
centrifugation and used for enzyme analysis.
Statistical analysis of the data was performed
by using SPSS (Version 18), using
independent paired t-test.
Results
Figure (1): A photomicrograph of a section in the liver of control group showing the portal area with
branches of portal vein (arrow head) and hepatic artery (double arrows).H&E X200
Figure (2):A photomicrograph of a section in the liver of MSG treated group II showing dilated congested
central vein (CV) and blood sinusoids (s) [H&E ×400].
Figure (3): A photomicrograph of a section in the liver of MSG treated group II showing that most of the
peripheral hepatocytes (thin arrows) appear with cytoplasmic vacuolization.[H&E ×400].

Effect of L-Glutamic acid on histology and functions of Liver and kidney Snoor Jalal Mustafa
of Rats and Protective Role of Zingibar Officionale
Diyala Journal of Medicine 54 Vol.11, Issue2,December2016
Figure (4):A photomicrograph of a section in the liver of MSG treated group II showing enlarged portal
area with branches of dilated portal vein (pv), congested hepatic artery (A), cellular infiltration (*) and
numerous bile ductules (thin arrows).[H&E ×400].
Figure (5):A photomicrograph of a section in the liver of MSG and ginger treated group IIІ showing
preservation of nearly normal hepatic lobular architecture with the presence of slightly dilated congested
central vein (cv) and blood sinusoids (curved arrows) with few cellular infiltration (*) [H&E ×400].
Figure (6): A photomicrograph of a section in a control rat’s kidneyshowing normal glomerular tuft (g) in
the cortex and tubules (t). (H & E X40)

Effect of L-Glutamic acid on histology and functions of Liver and kidney Snoor Jalal Mustafa
of Rats and Protective Role of Zingibar Officionale
Diyala Journal of Medicine 55 Vol.11, Issue2,December2016
Figure (7): A photomicrograph of a section in a control rat’s kidney showing renal corpuscle formed of
glomerulus (g) surrounded with Bowman’s capsule and preserved renal space(*).Note the proximal (P)and
distal (D)convoluted tubules(H&E X100).
Figure (8): A photomicrograph of a section from the kidney of a rat from groupII.Someglomeuli showing
shrinkage (arrow) and the others showing swelling with partial loss of the Bowman's spaces (g).(H & E
X100).
Figure (9): A photomicrograph of a section from the kidney of a rat of MSG group showing monocellular
inflammatory cell infiltrates (I). Tubular dilatation (D), hyaline casts (C) andvacuolization of tubular cells
are also observed (arrows) (H & EX200).

Effect of L-Glutamic acid on histology and functions of Liver and kidney Snoor Jalal Mustafa
of Rats and Protective Role of Zingibar Officionale
Diyala Journal of Medicine 56 Vol.11, Issue2,December2016
Figure (10):A photomicrograph of kidney section form a rat of MSG group Ishowing severe shrinkage of
the glomerulus (arrow) with increased in Bowman's spaces (*). There is also dilated congested interstitial
blood vessels and interstitial hemorrhage in the inter tubular spaces (crossed arrows). (H &E X400).
The biochemical parameters were
significantly higher in the MSG group II than
control groups (P < 0.0001) In group treated
with combination of MSG and Ginger the
results were.
Table (1): Comparison of means of liver function test parameters between Groups I, II, and III.
Group I
Mean±SD
Group II
Mean ±SD
Group III
Mean ±D
Bilirubin
0.1667±0.02108
0.1667±0.02108
0.2000±0.00000
Cholesterol
63.3333±1.28236
57.6667±1.97765
*↓
45.8000±1.39284
*↓
AST
234.00±1.57056
153.17±1.35195*↓
225.00±1.46059*
↓
ALT
57.6667±0.91894
33.5000±0.99163*↓
53.8333±0.98036
*↓
Albumin
1.3667±0.02108
1.2667±0.02108*↓
1.3333±0.02108
Table (2): Comparison of means of renal function test parameters betweenGroups I, II, and III.
Group I Mean±SD
Group II Mean ±SD
Group III Mean ±SD
Uric acid
1.3000±0.03651
2.6667±0.10220*↑
1.6833±0.04773*↑
Urea
15.6667±0.21082
17.5000±0.42817*↑
15.1667±0.30732
creatinine
0.7667±0.02108
0.9000±0.03651*↑
0.7333±0.03333
Na
133.83±0.60093
147.33±0.76012*↑
138.50±0.42817*↑
K
5.3000±0.09661
6.4167±0.10138*↑
5.6333±0.09545*↑
Cl
114.67±0.42164
102.67±0.88192*↓
110.00±0.57735*↓
Table (3): Comparison of means of liver function test betweenGroup II and III.
Group II Mean ±SD
GroupIII Mean ±SD
Bilirubin
0.1667±0.02108
0.2000±0.00000
Cholesterol
57.6667±1.97765 *↑
45.8000±1.39284*↓
AST
153.17±1.35195*↓
225.00±1.46059*↑
ALT
33.5000±0.99163*↓
53.8333±0.98036*↑
Albumin
1.2667±0.02108
1.3333±0.02108

Effect of L-Glutamic acid on histology and functions of Liver and kidney Snoor Jalal Mustafa
of Rats and Protective Role of Zingibar Officionale
Diyala Journal of Medicine 57 Vol.11, Issue2,December2016
Table (4): Comparison of means of renal function test between Group II and III.
Group II Mean ±SD
Group III
Mean ±SD
Urea
17.5000±0.42817*↑
15.1667±0.30732*↓
creatinine
0.9000±0.03651*↑
0.7333±0.03333↓
Na
147.33±0.76012*↑
138.50±0.42817*↓
K
6.4167±0.10138*↑
5.6333±0.09545*↓
Cl
102.67±0.88192*↓
110.00±0.57735*↑
Discussion
Monosodium Glutamate is one of the
most extensively researched food additives in
the world [9,12].Results of studies continue
to support the finding that at levels normally
consumed as flavor enhancer, MSG is safe
for the general population [6].Oxidative
stress is caused by excessive production or a
decreased elimination of free radicals in
cells, the majority of which are oxygen
radicals and other reactive oxygen species
(ROS)[6] .The abundance of long-chain
polyunsaturated fatty acids in the
composition of renal lipids makes kidney
susceptible to damage by ROS[13].This
makes kidney tissues prone to damage by
different mechanisms such as the promotion
of lipid peroxidation, protein modification,
and DNA damage, leading to cell death
[7,12]. Accordingly, the involvement of ROS
has been reported in glomerular, tubular and
tubulo-interstitial alterations [8,9].
Inthis study the liver of experimental
animals showed changes in histological
pattern evidentby disruption of hepatic cords,
presence of inflammatory cells within and
around the centralvein with uneven sizes of
nucleus in hepatocytes. Few reports on
alteration in liverhistology and/or
biochemistry have been documented
although these studies used doses thatwere
way above the dose we chose for this study
[10,15,16]. Monosodium glutamate may have
acted as toxins to the hepatocytes, thereby
affecting their cellular integrity and causing
defect in membrane permeability and cell
volume homeostasis. The atrophic and
degenerative changes observed in this
experiment may have been caused by the
cytotoxic effect of MSG on the liver. This
obviously will affect the normal
detoxification processes and other functions
of the liver.
Kidney microanatomy in group II that
received MSG compared to control showed
dilatation of the Bowman’s space,
contraction of the renal glomerulus
andhypercellularity which are in keeping
with renal injury, this corroborates results of
studiescarried out in 2007 by Eweka [1] .He
investigated the effects of MSG on the
kidney of adultWistar rats given 3g and 6g of
MSG thoroughly mixed with growers mash
for the period offourteen days, results of
kidney microanatomy showed varying
degrees of cytoarchitecturaldistortion and
reduction in the number of renal corpuscles
in the treated groups which was atvariance
with that of the control group.
Monosodium glutamate can induce
changes in the renal cytoarchitecture,
increase glomerular hyper-cellularity,
infiltration of inflammatory cells in the renal
cortex, edema of tubular cells, and eventually
degeneration of renal tubules [17-18]. These
results exactly confirm the results that we
found.The formation of ROS in the kidney
exposed to MSG was seen as a major
contributor to their nephrotoxic effects
leading to cellular and functional damage
[13]. MSG supplementation either by
injection or oral intake has been shown to
alter renal antioxidant system markers,
including lipid peroxidation byproducts and

Effect of L-Glutamic acid on histology and functions of Liver and kidney Snoor Jalal Mustafa
of Rats and Protective Role of Zingibar Officionale
Diyala Journal of Medicine 58 Vol.11, Issue2,December2016
kidney function in rats (15,17)]. In our study
the rats in group III which treated with MSG
and Ginger showed better results in their
liver and renal function tests and their
cytoarchitecture had been less affected, this
has occurred because of antioxidant role of
Ginger.Moreover, some studies have found
the ameliorating effect of vitamin C, E, and
Quercetin a plant pigment on MSG-treated
kidneys(17, 19, 20). The mechanism whereby
these antioxidants exert such effects is yet to
be fully elucidated. However, these
antioxidants seem to play a key role against
renal inflammatory responses through a
diminution of the activity of inflammatory
enzymes [1]. These important findings add
further prospective to the therapy of MSG-
induced renal oxidative stress using
antioxidants.However, altered kidney
function and pathology but not the renal
stones were reported by Paul et al. (2012)
after 6 months of oral MSG treatment with
higher dose (20). Cellular infiltration was
noticed in the portal areas in liver sections of
group ІІ in this study. These observations
were in accordance with the results obtained
by other studies that referred cellular
infiltration to ROS production which
indirectly regulate chemokine receptor
expression and promote cytokine IL-6 and
IL-8 which are key modulators of
inflammatory response(21,22). Other
investigators considered that cellular
infiltration as a prominent immune response
of the body tissues by movement of fluids
and leukocytes from the blood into the
extravascular tissues(23,24).
The effects observed in both the liver and
kidneys could have occurred because
theseorgans are involved in the metabolism
of glutamate or as in another study it may be
dueparticularly in the liver exacerbation of
trans-fat induced fatty liver disease in rats by
amechanism that includes increased central
adiposity and alterations in both hepatic
andwhite adipose tissue gene
expression [25, 26].
In conclusion, ginger can decrease the
damage tolivercells from oxidative damage
induces by MSG, and itis dependent on their
antioxidant effects.
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
 


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 


djm.diyala@yahoo.com, djm@medicine.uodiyala.edu.iq
 