https://doi.org/10.52973/rcfcv-e34347
Received: 05/11/2023 Accepted: 22/12/2023 Published: 18/03/2024
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Revista Científica, FCV-LUZ / Vol. XXXIV, rcfcv-e34347
ABSTRACT
This study was conducted to investigate the protective effects of
Betanin active ingredient in red beetroot plant (Beta vulgaris) in
elderly rats exposed to chronic toxicity of monosodium glutamate
(MSG). A total of 48 elderly rats were randomly divided into 4 different
deep anesthesia. Total antioxidant capacity (TAC), total oxidant
capacity (TOC), paraoxonase (PON), thiol, malondialdehyde (MDA),
and nitric oxide (NO) levels were investigated in rat blood serum using
the spectrophotometric method. Oxidative Stress Index (OSI) was
calculated by dividing TOC by TAC. Total bilirubin was measured with
the colorimetric method using an ELISA kit. Liver tissues were stained
with hematoxylin–eosin (HE) for histopathological examination. The
difference in serum levels of TAC, TOC, OSI, PON, MDA, and thiol was
P<0.05). The difference in
between the groups (P>0.05). The analysis of histopathological
MSG group and almost normal histological appearance in the
MSG+Betanin group. This study demonstrated that betanin could
increase the antioxidant effect and reduce the histopathological
damage caused by MSG.
Key words: Monosodium glutamate; aged rats; biochemistry;
betanin; histopathology
RESUMEN
Beta
grupos (P
P>0,05).
or el MSG.
Palabras clave:
Investigation of potential protective effects of Betanin on experimental
Monosodium Glutamate–induced toxicity in elderly rats
Investigación de los posibles efectos protectores de la Betanina sobre la toxicidad
experimental inducida por Glutamato Monosódico en ratas ancianas
Gurkan Baytar
1
, Tuncer Kutlu
2
, Serdal Ogut
3
*
1
Aydin Adnan Menderes University, Institute of Health Sciences, Aging Health and Care Interdisciplinary. Aydin, Türkiye.
2
Hatay Mustafa Kemal University, Faculty of Veterinary Medicine, Department of Veterinary Pathology. Hatay, Türkiye.
3
*Corresponding Author: serdalogut@yahoo.com
Protective effects of Betanin on Monosodium Glutamate toxicity in rats / Baytar et al. _____________________________________________
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INTRODUCTION
resulting from the increase in reactive oxygen species (ROS) formed
1, 2]. ROS have different chemical
structures such as hydroxyl radical, superoxide radical (O
2
·
–
), and
hydrogen peroxide. Numerous experimental data have shown an
increase in intracellular oxidative stress during aging. This is partly
due to the gradual reduction of intracellular ROS scavenging during
the aging pr3].
4, 5] causes
6]. However,
MSG has several adverse effects on various organs, including the
57, 87, 9]. The study by Nnadozie et al.7]
administering MSG Wistar albino rats for one year to observe mortality,
biochemical parameters of the MSG group compared to the control
7]. In the study by Hazza et al.10], MSG was administered to
Wistar albino rats (Rattus norvegicus) for 30 days. The study found
that MSG caused oxidative s10]
antioxidants they contain. Red beetroot is one of these important
11, 12]. A study investigating the protective properties
of betanin on many diseases in rats revealed positive results by
13].
Antioxidants are chemical substances that inhibit or delay undesirable
oxidation reactions. They prevent cell damage by presenting their
electrons to free radicals. Antioxidants protect the substrate from
irreversible damage and prevent its oxidative conversion via reactive
species. Antioxidants in food delay, control, or prevent oxidation and
24
H
26
N
2
O
13
) is a good free radical
14]. In line with this information,
recent studies have emphasized the antioxidant property of Betanin.
In summary, the biochemical changes that occur with aging and the
additional burden of MSG, which is a food additive, lead to pathological
liver damage, an increase in oxidant parameters, and a decrease in
antioxidant parameters. Likewise, Betanin, which has an antioxidant
effect, prevents or decreases liver damage, decreases oxidant
parameters, and increases antioxidant parameters. However, these
changes have not yet been fully elucidated in elderly rats (Wistar Albino).
This study aimed to investigate the effects of Betanin on
Monosodium Glutamate–induced toxicity in blood serum and liver
in elderly rats with biochemical and histopathological methods.
MATERIALS AND METHODS
Ethics and animals
The present study was reviewed and approved by the institutional
Experiments Local Ethics Committee in January 2020 (IRB Approval
Number: 64583101/2020/05).
Forty–eight male wistar albino rats of minimum 15 month old and
about 350 g body weight (rat –wistar albino– Hatay Mustafa Kemal
acclimatized for 10 days (d) before the beginning of the experiment.
All rats fed standard rat diet, allowed to water ad libitum, and kept
under normal daylight/dark cycle and room temperature during study.
Experimental design
Rats were weighed and assigned into four experimental groups. It
is planned to have 12 rats in each group against the risk of death of
aged rats. Application took 28 d.
guiding principles for the ethical use of animals in rese15].
The present study was reviewed and approved by the institutional
Experiments Local Ethics Committee in January 2020 (IRB Approval
Number: 64583101/2020/05).
MSG was obtained from Sigma Company Germany. Betanin was
obtained from Sigma Aldrich.
○
Control group: Fed standard rat diet, allowed to water ad libitum
○ MSG group: 120 mg·kg
-1
·day
-1
MSG was administered by gavage
○
Betanin group: 20 mg·kg
-1
·day
-1
betanin was administered by gavage
○ MSG+Betanin group: 120 mg·kg
-1
·day
-1
MSG and 20 mg·kg
-1
·day
-1
Betanin was administered by gavage
Analyses
ELISA kits (Rel Assay Diagnostic–Turkey) were used for biochemical
16]. The colorimetric method is based on the measurement
with a reagent. Micro pellet reader devices show parallelism in many
color and severity of the reac17].
TAC and TOC measurements were performed by means of kits (Rel
1, 18]. MDA in rat blood was
determined according to the spectrophotometric method described by
Placer et al.19] and the results obtained were expressed as nmol·mL
-1
19
described by Navarro–Gonzalves et al.20]. Paraoxonase activities
measurements were performed in the absence (basal activity) and
presence of NaCl (salt–stimulated activity). Paraoxonase activity was
-1
s21].
Pathological evaluation
to 4–mm–thick tissue sections and washed overnight with running
tap water. After passing through a graded series of alcohol (50, 80,
the tissue blocks that were prepared in this way with a microtome
bath set at a temperature of 37°C, and then taken to slides and dried
in xylol and immersed through alcohol series of 100, 96 and 80, and
70%, they were stained with Hematoxylin Eosin (HE). They were again
A B
C D
FIGURE 1. Hepatic Histopathological Findings. A) Normal histological structure
of the liver, Control Group, HE. B) Hydropic degeneration of hepatocytes (stars),
MSG, HE. C) Focal mononuclear cell inltration of the portal tract (arrows), MSG
Group, HE. D) Normal histological structure of the liver, MSG+Betanin Group, HE
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passed through a graded series of alcohol (80, 96 and 100%) and xylol
and closed with a coverslip with the help of a glue substance (Entellan).
After examination under a light microscope (Olympus CX31, Münster,
Germany), microphotographs (Olympus DP12, Hamburg, Germany)
were taken. Histopathological changes were scored according to
the following criteria: Grade 0, histopathological changes below 5%
of the entire area; Grade 1, mild histopathological changes in 5–33%
of the entire area; Grade 2, moderate histopathological changes in
33–66% of the entire area; Grade 3, severe histopathological changes
in more than 66% of the entire area22, 23].
Statistical evaluation
package. The test for conformity to normal distribution was evaluated
with the Shapiro Wilk Test. One–way ANOVA method determined
P<0.05). Post–hoc test (Duncan)
cant data.
RESULTS AND DISCUSSION
Biochemical Results
the groups in terms of TAC, TOC, OSI, PON, thiol, and MDA (P<0.05).
The comparison of the MSG group with the Betanin group showed
Betanin groups (P<0.05). This result supports the antioxidant property
of Betanin. The comparison of the MSG group with the Betanin group
values in the MSG groups (P<0.05). This result supports the oxidant
property of MSG. The comparison of the control group with other
of total bilirubin and NO (P>0.05).
Pathological results
In the control and Betanin groups, the liver had a normal histological
structure (FIG. 1A). Mild hydropic degeneration, which was uncommon
in hepatocytes, was noted in the MSG group (FIG. 1B). In addition, focal
foci of mononuclear
(FIG. 1C). The MSG+Betanin group had an almost normal histological
appearance (FIG. 1D).
TABLE I
Serum TAC, TOC, OSI, Total Bilirubin, PON, Thiol, NO and MDA Levels
Control
X
̄
± SD
MSG
X
̄
± SD
Betanin
X
̄
± SD
MSG+Betanin
X
̄
± SD
P–value
TAC
(mmol Trolox equiv·L
-1
)
2.00 ± 0.20
a
1.47 ± 0.13
ab
1.94 ± 0.13
b
1.77 ± 0.18
b
P<0.05
TOC
(μmol H
2
O
2
equiv·L
-1
)
9.93 ± 1.33
a
14.14 ± 0.16
ab
10.05 ± 0.93
bc
12.05 ± 1.16
abc
P<0.05
OSI
[(TOC/TAC)×100]
50.24 ± 9.63
a
96.71 ± 10.40
ab
51.89 ± 6.49
bc
68.62 ± 8.97
abc
P<0.05
Total Bilirubin
(U·L
-1
)
0.029 ± 0.012 0.031 ± 0.014 0.033 ± 0.013 0.024 ± 0.011
P>0.05
PON
(U·L
-1
)
297.69 ± 13.92
a
222.87 ± 32.18
ab
300.00 ± 21.38
b
280.19 ± 13.63
b
P<0.05
THIOL
(μmol·L
-1
)
283.63 ± 10.35
a
301.05 ± 10.74
ab
287.66 ± 8.90 279.49 ± 8.64
b
P<0.05
NO
(µmol·L
-1
)
13.68 ± 4.65 17.07 ± 7.70 18.50 ± 5.40 19.74 ± 5.15
P>0.05
MDA
(mmol·L
-1
)
4.07 ± 0.58
a
6.07 ± 0.95
ab
4.43 ± 0.74
b
5.06 ± 0.49 P<0.05
P<0.05 (statistically signicant). P>0.05 (statistically non signicant).
a,b,c
: groups with the same letters on the same
line are dierent from each other
The comparison of the control group with the MSG group
P<0.05), while the comparison
difference (P>0.05). When the MSG group and the betanin group
P<0.05).
by MSG (FIG. 2).
FIGURE 2. Mononuclear Cell Inltration
FIGURE 3. Hydropic Degeneration of Hepatocytes
Protective effects of Betanin on Monosodium Glutamate toxicity in rats / Baytar et al. _____________________________________________
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The comparison of the control group with the MSG group showed
P<0.05), while the comparison of the control
(P>0.05). Likewise, the comparison of the MSG group with the Betanin
P<0.05). Hydropic degeneration
of hepatocytes was caused by MSG (FIG. 3).
Exposure to the food additive MSG with age can further increase
oxidative stress. Moreover, the elimination of oxidative stress or the
reduction of its effects can be ameliorated with betanin, which is also
used in the food industry and has an antioxidant effect. Biochemical
and pathological investigation of these changes is important in terms of
gaining new perspectives. Accordingly, in this study, it was investigated
TAC, TOC, Thiol, PON, NO, MDA, and total Bilirubin expressions in the
was examined the OSI obtained by dividing TOC by TAC.
Thiols, which are organic sulfur derivatives containing sulfhydryl
(–S–S–) bond formation, easily react with oxygen–containing free
25]. Paraoxonases (PONs) comprise a gene
family consisting of three members (Paraoxanase 1, 2, 3) with diverse
26]. Nitric oxide (NO) functions as an oxidative biological signaling
27]. MDA is an important biomarker to
28]. In a study on Wistar Albino
rats, MSG was administered orally, and MDA values were examined
as an indicator of oxidative stress. MDA values were statistically
29]. MDA levels and their
effects on liver morphology were evaluated in mice fed with a standard
diet or a high–fat diet. MSG was found to have effects on oxidative
30, 31]. In the present study, the MDA
serum levels showed an increase in all groups compared to the control
group. However, only the increase in the MSG group was statistically
er tissue.
A study by Da Silva et al.32] evaluated the effect of betanin intake
for 20 d on oxidative stress in Wistar albino rats. The results of the
study showed reductions in MDA levels of betanin–treated rats and
32]. In a study
33].
In the present study, the highest MDA value was observed in the MSG
group. This result clearly shows that MSG causes lipid peroxidation in
rats. Other studies also support this result. Furthermore, the current
response) and degenerative changes in the liver induced by MSG.
Bilirubin is constantly produced by liver, spleen, or bone marrow
macrophages with the breakdown of heme from senescent red blood
3435] found that MSG
35]. In the current study, the differences
P>0.05).
There was an increase in serum levels of total bilirubin in the MSG group
and the Betanin group compared to the control group. However, the
serum levels of total bilirubin in the MSG+Betanin group compared to the
control group. However, the decrease was not statistically si
A study by Helal et al.36] designed to investigate its therapeutic
role in MSG–induced male reproductive system disorders in male rats
36
levels between the groups (P>0.05).
Aging is a physiological process characterized by molecular and
24].
Therefore, elderly people who are sensitive to external factors are
adversely affected by food additives. A better understanding of
biochemical parameters and how the liver is pathologically affected can
minimize oxidative damage. Accordingly, it is important to understand
the biochemical and pathological changes in aging metabolism. In this
context, it is even more important to use natural antioxidants in food
technology in order to detoxify the negative effects of food additives.
This study aimed to investigate the protective effects of Betanin against
the toxic effects of MSG in Elderly rats.
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There is no study in the literature administering MSG to elderly rats
and investigating the thiol levels. In the present study, the difference
(P<0.05). The comparison of the control group and other groups
increase in the MSG group. The comparison of the MSG group with
the Betanin and MSG+Betanin groups revealed a decrease in thiol
group. It is believed that the simultaneous administration of MSG
and Betanin may cause toxic effects.
In their study, Elbassuoni et al. found significantly lower TAC
37]. A study
investigating the effect of Betaine supplementation on cadmium–
induced oxidative impairment in rat kidney reported that Betanin
administration alleviated the reduction in TAC in cadmium+Betanin–
38]. The results of
the present study showed that the difference in TAC serum levels was
P<0.05). The comparison
of the control group and other groups in terms of serum levels of TAC
revealed a decrease in the TAC serum level of all groups. The lowest
TAC value was found in the MSG group. Following the control group,
the highest TAC value was found in the Betanin group. The result of
the lowest TAC level in the MSG group suggests that MSG may have an
oxidant effect. Betanin is thought to increase antioxidant capacity.
In addition, when administered with MSG, it is believed to reduce the
oxidant effect of MSG. In the present study, the difference in serum
P<0.05).
The highest TOC value was observed in the MSG group. Following the
control group, the lowest TOC value was found in the Betanin group.
Due to the oxidant effect of MSG, the highest TOC value was noted
in the MSG group. Due to its antioxidant properties, Betanin reduces
the oxidant effects of MSG. The results of the comparison of the
TOC values in the MSG+ Betanin group and the MSG group support
this result. The difference in serum levels of OSI was statistically
P<0.05). The highest OSI value was
observed in the MSG group. Following the control group, the lowest
OSI value was noted in the Betanin group. Considering the OSI values,
the antioxidant property of Betanin has been proven once again.
Serum levels of thiol are presented in TABLE I.
In a cell culture model study by Esatbeyoglu et al.39], which
aimed to systematically evaluate Betanin's free radical scavenging
activity by electron spin resonance spectroscopy and spin capture,
the antioxidant activity of Betanin was determined in terms of
transactivation in the cell culture m39].
In the present study, the difference in serum levels of PON was
value was observed in the Betanin group. The lowest PON value was
noted in the MSG group. These results show that Betanin increases
the activity of PON, which is an antioxidant. On the contrary, MSG with
toxic and oxidant properties reduces the activity of PON.
The present study revealed the toxic effects of MSG once again.
The comparison of the MSG groups with the control group showed
lower TAC and PON levels in the MSG group. The histological
comparison of the MSG groups with the control group revealed
that the liver tissue of the control group was normal, while the MSG
in hepatocytes. On the other hand, the Betanin groups had a normal
histological structure of the liver.
CONCLUSION AND IMPLICATIONS
MGS is widely consumed as a food additive in the World. The
results of this dissertation study once again revealed the negative
health effects of MSG. Therefore, more attention should be paid
to the consumption of foods containing this food additive. Foods
containing low amounts of MSG or not containing MSG should be
consumed. The results of the present dissertation study support the
antioxidant property of Betanin. These results indicate that Betanin
reduces the oxidant effects of MSG. Accordingly, it is believed that
adding red beetroot to diets will have positive effects. The elderly
are known as a vulnerable group in society. The results of this study
on elderly rats once again demonstrated the sensitivity of this group.
The elderly need to be more careful when consuming foods containing
MSG. MSG–free foods should be chosen. Betanin can be preferred as
a nutrient enhancer in the food industry.
There is a need for further studies to determine the antioxidant
effects of the long–term use of Betanin at different doses on the
aging process.
ACKNOWLEDGMENTS
some of the findings were presented at the scientific meeting
Gerontology Studies Application and Research Center 1. International
Congress Of Gerontology, 18–20 March 2022).
Declaration of conicting interests
to the research, authorship, and/or publication of this article.
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