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Imperial Journal of Interdisciplinary Research (IJIR)

Vol-2, Issue-12, 2016


ISSN: 2454-1362, http://www.onlinejournal.in

Physical and Clinical Changes Induced by Monosodium


Glutamate in Swiss Albino Mice and its Counter-Action
by the Water Extract of Withania Somnifera Root
Genevia Fatima Mercy. V*, Geegi. P.G.
Department of Biochemistry, St. Josephs College, Tiruchirappalli.
Abstract: In the recent times, the hidden additives of fast effects against the major complications of MSG
foods have created a larger impact on health issues consumption. Oral ingestion of Ashwagandha was
where, they are found to induce metabolic changes, able to reduce the triglyceride levels in serum to its
leading to several disorders. Monosodium Glutamate is normal level. In persons with metabolic syndrome,
the most commonly used additive, which can lead to the supplementation of 400mg of Ashwagandha extract
development of obesity and other neurodegenerative thrice a day for 30 days is able to reduce triglycerides
disorders. This forms the background of this study. A [5][6][7]
by approximately 12% relative to baseline. In
total of 24 Swiss albino mice were taken and were diabetes induced rats, the abnormalities observed in
divided into 4 groups with 6 animals each. Test group the serum cholesterol level appears to be fully
was administered with MSG at 4g/Kg (oral). Test + normalized with ingestion of 1,000mg/kg of
treatment group was initially administered with same
Ashwagandha extract. In rodent studies,
quantity of MSG followed by the oral administration of
Ashwagandha is noted to better the lipoprotein profile
Withania somnifera water extract (10g/Kg). By the end
with a significant increase in HDL and more notable
of 40 days the animals were sacrificed and the serum
reductions in LDL and total cholesterol. The
levels of triglyceride, glucose, total protein and
reduction in LDL cholesterol actually appears to be a
cholesterol were estimated. Results of these estimations
per se mechanism of action rather than just fixing a
were recorded and presented with suitable graphical
metabolic abnormality, and occurs in normal rats as
representations. From the results obtained it becomes [8]
evident that MSG is capable of inducing metabolic well as those with metabolic ailments. At the same
changes and obesity which can be effectively treated by time 200-400mg/kg of the aqueous root extract has
Withania somnifera. been noted to normalize blood glucose in
streptozotocin induced diabetic rats after five weeks
Key Words: Withania somnifera, obesity, by 76-89%, which correlated with improvements in
neurodegenerative disorder, metabolic changes. oxidative biomarkers and pancreatic cell structure.
[6][9]
It also has the potential to induce apoptotic cell
1. Introduction death in fat cells, and hence it has the ability to revert
[10]
Recent researches have evidently shown that a back obesity which is being induced by the MSG .
common additive of fast foods known as Monosodium This is supported by a study where untrained men
glutamate (MSG) induces obesity in laboratory animals were put on a periodized training program for 8
by damaging the arcuate nucleus of hypothalamus. This weeks where they were given with 300 mg of a
arcuate nucleus is considered to be the appetite commercially available ashwagandha extract daily.
regulation centre. Any damage in this region will result After the study period it was found that the persons
in the development of resistance to the hormone leptin who took Ashwagandha extract lost 2% more body
[1] fat than the placebo group as measured by
which controls the feeling of satiety. Many research [11]
works have shown that some parts of the brain are bioelectrical impedance. With these studies as
extremely sensitive to a group of chemicals called base we carried out this study where we tested W.
excitotoxins. MSG is one of the excitotoxin, which somnifera extract in against the physical and clinical
causes damage to brain cells when its amount in blood changes that are induced by Swiss Albino Mice.
[2]
goes high. This study is aimed at bring out an 2. Materials and Methods
effective counteraction for these excitotoxins. A
promising outcome was expected from a plant named 2.1 Withania somnifera extract preparation
Withania somnifera which is a highly esteemed
Commercially available powdered form of the plant
medicinal herb in Ayurveda which is commonly known
[3] was brought from the local grocery on February 3rd
as Ashwagandha . It has been traditionally used as an 2016. Thereafter, 50g of powder was suspended in 500
analgesic, astringent, anti-spasmodic, and immuno ml of boiling water and thoroughly mixed and kept as
stimulant while being used to treat inflammation, such for 48 h in dark (percolation method). The
cancer, stress, fatigue, diabetes, and cardiovascular extract was then filtered and concentrated to obtain the
[4] Solid residue which was refrigerated until further use.
complications . There were many evidences in which
Ashwagandha was reported to have counteractive

Imperial Journal of Interdisciplinary Research (IJIR) Page 1264


Imperial Journal of Interdisciplinary Research (IJIR)
Vol-2, Issue-12, 2016
ISSN: 2454-1362, http://www.onlinejournal.in

WS extract of lower concentrations were prepared by 3. Results


dilution of the stock with hot water.
3.1. Physical & Behavioural activity and
2.2 Experimental Procedure weight gain
A total of 24 (12 female and 12 male) Swiss
While the physical activity and eating was
Albino mice were brought from Sri Venkateshwara
observed the test group animals showed hyper
Enterprises, Bangalore, India which were about one
activeness in the first 10 days of study and their
month old and were housed in the temperature
activeness reduced to idleness in the next 20 days
controlled Animal house at St. Josephs college,
and at the same time these animals were found to
Tiruchirappalli. All the experimental procedures were
consume more feed than the other group of
carried out on the basis of IAEC guidelines. The
animals. Of the four groups the test group,
average weight of these animals were recorded as 23g
administered with MSG showed a rapid weight
on the first day of study. The animals were allowed to
gain, leading to the development of obesity. Some
acclimatize for a period of ten days. Then they were
of animals in the group II became ferocious and
divided into four groups with 6 animals in each (with 3
attacked the other animals kept within the cage.
female and 3 male mice).
Hence these animals were isolated for 2 days and
GROUP I: Control group animals were fed when they were put back into the group, they
with clean water and commercially available mice showed the same ferocious behaviour, and they
feed. were found to have higher degree of obesity. Even
though they were obese they were hyperactive. The
GROUP II: Test group were fed with mice feed average weights of all the groups were measured
and water similar to the control group along with the for every 10 days. The comparison of each group
oral administration of MSG for 40days at 4g/kg/day. on weight gain showed that the weight of mice
belonging to Group 2 (Test) was obese and the rest
GROUP III: Test + Treatment group animals were normal (Fig.1)
were fed with MSG (4g/kg/day) orally for 10 days
followed by the oral administration of Withania Figure. 1: Average Weight of Mice at every ten days
somnifera water extract at 10g/Kg/day for 30 days. in each group

GROUP IV: Treatment control group mice


were administered with Withania somnifera extract at
10g/Kg/day for 40 days.
The average weights of animals in
correspondence with each group were calculated at a
regular interval of 10 days and were recorded. A
graph was plotted using the observed weights of all
the animals during the study period. At regular
intervals the physical and behavioural activities of
th
the animals were observed. On the 40 day of our
work all the mice were sacrificed and blood samples
and tissue samples of brain, liver and kidney were
collected and preserved for future use. Serum was
separated and was estimated for the levels of total
protein, glucose, triglycerides, and cholesterol. 3.2. Estimation of Triglycerides
Statistical analysis were done with the results
obtained from the estimations. When the serum levels of triglyceride were
estimated using laboratory protocol and there was a
2.3 Statistical Analysis significant increase observed in the group 2
animals. While the animals belonging to other
The values obtained from the quantitative analyses groups showed levels of triglycerides within the
of serum biomolecules were recorded, tabulated and normal range. The mean and standard deviation
represented as Mean Standard deviation. To compare values were calculated and tabulated (Table. 1) and
the values obtained for the four different groups for were plotted in a graph (Fig. 2)
differences for each estimations t test was calculated
and significances were determined

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Imperial Journal of Interdisciplinary Research (IJIR)
Vol-2, Issue-12, 2016
ISSN: 2454-1362, http://www.onlinejournal.in

Table 1: Estimation of Triglyceride (mg/dl) Mean and standard deviation were calculated
and tabulated (Table. 2) and were represented
S.No Groups Mean SD diagrammatically using a graph (Fig. 3)
1. Control 91.945 2.651
2. Test 221.43 22.01 Table 2: Estimation of Glucose (g/dl)
3. Test+Treatment 98.96 6.024
S.No Groups Mean SD
4. Treatment control 88.42 1.499
1. Control 73.635 5.176
Figure. 2 Estimation of Triglycerides 2. Test 186.745 5.253
3. Test+Treatment 147.86 14.340
4. Treatment control 83.185 1.675

Figure. 3 Estimation of Glucose

The following results were obtained when the


groups were compared using ttests.
Group I vs. Group II
P Value and statistical significance:
The two-tailed P value is 0.0143. By
conventional criteria, this difference is considered to
be statistically significant. The following results were obtained when the
Group I vs. Group III groups were compared using ttests.

P Value and statistical significance: Group I vs. Group II

The two-tailed P value is 0.2707. By P Value and statistical significance:


conventional criteria, this difference is considered to be
not significant. The two-tailed P value is 0.0021. By
conventional criteria, this difference is considered
Group II vs. Group III to be statistically very significant.

P Value and statistical significance: Group I vs. Group III

The two-tailed P value is 0.0169. By P Value and statistical significance:


conventional criteria, this difference is considered to
be statistically significant. The two-tailed P value is 0.0204. By
conventional criteria, this difference is considered
Group II vs. Group IV to be statistically significant.

P Value and statistical significance: Group II vs. Group III

The two-tailed P value is 0.0135. By P Value and statistical significance:


conventional criteria, this difference is considered to
be statistically significant. The two-tailed P value is 0.0692. By conventional
criteria, this difference is considered to be
3.3. Estimation of Glucose statistically not significant.

When the serum was analysed for the Group II vs. Group IV
quantitative determination of glucose the results
P Value and statistical significance:
showed a peak increase of serum glucose levels in
the group 2 animals.

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Imperial Journal of Interdisciplinary Research (IJIR)
Vol-2, Issue-12, 2016
ISSN: 2454-1362, http://www.onlinejournal.in

The two-tailed P value is 0.0014. By


conventional criteria, this difference is considered to The two-tailed P value is 0.0692. By
be statistically very significant. conventional criteria, this difference is
3.4 Estimation of Cholesterol considered to be statistically significant.

Cholesterol levels of serum were analysed in Group II vs. Group IV


the serum samples of the study animals. Cholesterol
levels are the clear indicators of development of P Value and statistical significance:
obesity. The estimation was carried out using Zaks The two-tailed P value is 0.0046. By
protocol for the determination of serum cholesterol conventional criteria, this difference is considered
level. From the results obtained mean and standard to be statistically very significant.
deviation values were calculate and tabulated (Table.
3) and were represented using a graph. (Fig. 4) 3.5. Estimation of Total protein
Table 3: Estimation of Cholesterol (mg/dl) The serum samples of the animal models used
for the experiment were analysed for their total
S.No Groups Mean SD
protein levels. The mean and standard values of
1. Control 101.39 5.784 the determined levels are calculated for each
2. Test 202.43 9.418 group and were tabulated (Table. 4) and were
graphically represented as shown in Fig.5
3. Test+Treatment 119.40 3.390
4. Treatment control 100.21 2.927 Table. 4: Estimation of Total Protein

S.No Groups Mean SD


Figure. 4 Estimation of Cholesterol 1. Control 3.45 0.353
2. Test 7.30 0.141
3. Test+Treatment 5.55 0.353
4. Treatment control 5.15 0.353

Figure. 5 Estimation of Total Protein

The results of the statistical analysis of


comparison of groups are as follows:

Group I vs. Group II


P Value and statistical significance:
The two-tailed P value is 0.0059. By
conventional criteria, this difference is considered The results of the statistical analysis of
to be statistically very significant. comparison of groups are as follows:
Group I vs. Group III Group I vs. Group II
P Value and statistical significance: P Value and statistical significance:
The two-tailed P value is 0.0072. By The two-tailed P value is 0.0048. By conventional
conventional criteria, this difference is considered criteria, this difference is considered to be
to be statistically very significant. statistically very significant.
Group II vs. Group III Group I vs. Group III
P Value and statistical significance: P Value and statistical significance:

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Imperial Journal of Interdisciplinary Research (IJIR)
Vol-2, Issue-12, 2016
ISSN: 2454-1362, http://www.onlinejournal.in

status were between 2.10 and 2.75 greater than for


The two-tailed P value is 0.0271. By conventional [12]
nonusers.
criteria, this difference is considered to be
statistically significant. In another research the MSG fed mice had
developed obesity with visceral fat deposition
Group II vs. Group III regardless of the dietary regimen. Both MSG treated
groups did not show significant lower body weight
P Value and statistical significance: regardless of diet restriction at both evaluation
[13]
The two-tailed P value is 0.0692. By conventional points.
criteria, this difference is considered to be statistically The animals that are kept under group 3 are the one
not significant. which were tested with the same quantity of
Monosodium glutamate as group 2 animals but they
Group II vs. Group IV were treated with the oral administration of the water
P Value and statistical significance: extract of Withania somnifera (10mg/g) during the
second half of the study period. This treatment has
The two-tailed P value is 0.0228. By conventional shown a very good result were the obesity
criteria, this difference is considered to be statistically development was brought back to normal and the
significant. serum levels of triglyceride, cholesterol, glucose and
total protein were found to be normal, but were slightly
4. Discussion higher than the control group values. This alteration of
abnormal values to normal values is due to the effect of
A total of 24 test animals (Swiss Albino Mice) the plant extract of Withania somnifera.
were taken into study and they were divided into four
groups and each followed a different treatment.
This effect was studied in cell lines by Kataria
The first group was regarded as a control group Hardeep and colleagues, where they studied the
for which no drug or treatment was administered. The protective effect of water extract from the leaves of
serum levels of glucose, triglyceride, cholesterol and Withania somnifera on RA Differentiated C6 and
total protein remained normal as the metabolic IMR-32 Cells against Glutamate Induced
functions were normal. Healthy weight gain was
Excitotoxicity where the results showed that he
recorded.
ASHWEX pre-treatment inhibited glutamate induced
In case of group 2, the animals were tested with cell death and was able to revert glutamate induced
daily oral administration of Monosodium glutamate changes in HSP70 to a large extent. Furthermore, the
(4mg/g). These set of mice showed the development analysis on the neuronal plasticity marker NCAM
of obesity with increased abnormal levels of (Neural cell adhesion molecule) and its poly sialylated
Triglyceride, cholesterol, glucose and total protein. form, PSA-NCAM revealed that ASHWEX has
This may be due to the metabolic changes induced by therapeutic potential for prevention of
the ingestion of Monosodium Glutamate. Neurodegeneration associated with glutamate induced
[14]
excitotoxicity.
These results were facilitated by a research work
conducted to examine the association between MSG This result is also facilitated by many researches
intake and weight gain in humans. Researchers where W.somnifera was found to have anti-
[15][16]
conducted a cross-sectional study involving 752 healthy hyperlipidemic, anti-diabetic activities.
Chinese people ages 40-59. These people were randomly
Group 4 was considered as standard control group
sampled from three rural villages in north and south
to which daily oral administration of Withania
China. Forty-eight percent were women. The great
somnifera was given at 10mg/g concentration. Healthy
majority of the participants prepared their foods at home, weight gain was found. The levels of triglyceride,
without use of commercially processed foods. For the cholesterol, glucose and total protein in serum were
study, participants were told to add quantified amounts slightly higher than the control animals but they were
of MSG when preparing their foods. Eighty-two percent still found to be in the normal range. This increase may
of the participants were MSG users. Their average intake be due to the regulatory effects of the plant extract.
was 330 milligrams per day. After adjusting for
confounders including physical activity and total energy These health benefits of Withania somnifera is
intake, the researchers found that MSG consumption was being explicitly supported by a wide range of
positively related to increases in body mass index. studies where it is found to improve general health
Weight gain was significantly greater in MSG users than conditions which includes learning and
in nonusers. For the third of participants using the [17][18] [19][20]
memory sedation and sleep
highest amount of MSG, the odds of reaching overweight [21]
neurogenesis etc.,

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Imperial Journal of Interdisciplinary Research (IJIR)
Vol-2, Issue-12, 2016
ISSN: 2454-1362, http://www.onlinejournal.in

When the results were compared for the means, Exploratory study to evaluate tolerability, safety
the values were found to be statistically significant and activity of Ashwagandha (Withania somnifera)
between the groups 1 & 2, 2 & 3 and 2 & 4 but was in healthy volunteers. Journal of Ayurveda and
Integrative Medicine, 3(3): 111-114, July 2012.
insignificant between the groups 1 & 3 for triglyceride
[6] Datta A, Bagchi C, Das S, Mitra A, Pati AD.
estimation, which shows that Monosodium Glutamate Tripathi SK. Anti-diabetic and anti-hyperlipidemic
has an effect of triglyceride metabolism which was activity of hydro alcoholic extract of Withania
reverted by the plant extract of Withania somnifera. coaglans Dunal dried fruit in experimental rat models,
Journal of Ayurveda and Integrative Medicine, 4(2):
From the test for significance for the glucose showed 99-106, April 2013.
statistically significant difference between the groups 1 [7] Visavadiya NP, Narasimhacharya AV. Hypo-
& 2, 1 & 3 and 2 & 3 but was insignificant between 2 cholesteremic and antioxidant effects of Withania
& 4 which supports the hypothesis that MSG somnifera (Dunal) in hyper-cholesteremic rats.
induces hyperglycemia and W.somnifera has the Phytomedicine, 14(2-3): 136 142, February 2007.
ability to bring down the hyperglycemic state. [8] Visavadiya NP, Narasimhacharya AV.
Ameliorative Effects of Herbal Combinations in
For the estimation of cholesterol there was a Hyperlipidaemia Oxidative Medicine and Cellular
significant variation in almost all the group and no Longevity, 2011:18,
significance was identified. September 2011.
[9] Anwer T, Sharma M, Pillai KK, Khan G.
In the total protein estimation insignificant value Protective effect of Withania somnifera against
of difference between the means of group 2 & 3 oxidative stress and pancreatic beta-cell damage in
type 2 diabetic rats Acta Poloniae Pharmaceutica,
were observed and other groups show significant
69(6):1095-1101, December 2012.
variations.
[10] Park HJ, Rayalam S, Della-Fera MA, Ambati S,
Yang JY, Baile CA. Withaferin A induces
5. Conclusion apoptosis and inhibits adipogenesis in 3T3-L1
adipocytes, Biofactors, 33(2):137-148 2008.
From this work we could conclude that addition
[11] Wankhede S, Langade D, Joshi K, Sinha SR
of MSG in food affects the normal metabolic Bhattacharyya S. Examining the effect of
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obesity etc., and can increase the serum levels of strength and recovery: a randomized controlled
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result in the development of atherosclerosis and even Nutrition, 12:43, November 2015.
more serious problem. Hence it is not advisable to [12] He K, Zhao L, Daviglus ML, Dyer AR, Van Horn
add MSG as an ingredient in our daily diet. L, Garside D, Zhu L, Guo D, Wu Y, Zhou B,
Stamler J. Association of monosodium glutamate
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16(8):1875-1880, August 2008.
Withania somnifera commonly known as
[13] Fujimoto M, Tsuneyama K, Nakanishi Y
Ashwagandha which is also commercially available,
Salunga TL, Nomoto K, Sasaki Y, Iizuka S
can act as a perfect remedy for all the damages Nagata M, Suzuki W, Shimada T, Aburada M,
caused to the body by monosodium glutamate. It can Shimada Y, Gershwin ME, Selmi C. A dietary
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Imperial Journal of Interdisciplinary Research (IJIR) Page 1269


Imperial Journal of Interdisciplinary Research (IJIR)
Vol-2, Issue-12, 2016
ISSN: 2454-1362, http://www.onlinejournal.in

(Withania somnifera, Dunal) root, Indian Journal of


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