EFFECT OF CORN SILK, PUMPKIN SEEDS, AND
CREATINE ON OXIDATIVE STRESS OF MALE
EXERCISED RATS
Naeem M. Rabeh ; Mohammed H. Haggag
and Asmaa Z. Mohammed
Nutrition and Food Sci. Dept, Faculty of Home Economics, Helwan Univ., Cairo-Egypt
Key Words: Corn silk, Pumpkin seeds, creatine supplementation,
creatine kinase, lactate dehydrogenase, treadmill,
oxidative stress.
ABSTRACT
The present study was conducted to investigate the effect of corn silk,
pumpkin seeds and creatine supplementation on oxidative stress biomarker
as well as liver function and glycogen content of adult male exercised rats.
Fifty male albino rats were fed on basal diet for one week for adaptation.
During the adaptive week all rats (except negative control group) were
exposed to day after day forced treadmill running exercise for 10 min. Rats
were divided into two main groups, the first main group (n=10 rats) was fed
on basal diet only and served as negative control group. The second main
group (exercised rats) were divided into 4 subgroups (10 rats each) as
followed:-Subgroup 1: were fed on basal diet and had run for 10, 15, 20 and
25minutes (15 days for each time period), respectively and served as
positive control group. The Subgroups (2-4) as the same of positive control
subgroup and were fed on basal diet supplemented with 4% corn silk
powder, 4% pumpkin seeds powder and 4% creatine powder, respectively.
At the end of the experimental period (60 day), 5 rats of each subgroup were
exposed to the exhausting treadmill test and the run time was recorded, then
all experimental rats were scarified. Findings of these study prove that corn
silk and pumpkin seeds were a good source of flavonoid which help to
scavenge the oxidative stress substances, and improve liver functions , while
creatine recorded the largest glycogen quantity in gastrocnemius muscles
among supplemented groups, but it causing an elevation ammonia, and AST
levels on the group which exposure to exhaustive time test. It could be
recommended that corn silk and pumpkin seeds have the ability to eliminate
physical fatigue and prolong the training period without fatigue
INTRODUCTION
Biomarker discovery and validation is a critical aim of the medical
and scientific community. Research into exercise and diet-related
biomarkers aims to improve health, performance, and recovery in
military personnel, athletes, and lay
Persons. Exercise physiology research has identified individual
biomarkers for assessing health, performance, and recovery during
Egypt. J. of Appl. Sci., 35 (12) 2020 230-252
exercise training. However, there are few recommendations for
biomarker panels for tracking changes in individuals participating in
physical activity and exercise training programs (lee et al .,2017). The
diversity of nutrients or compounds from food factors or medical herbs
could be investigated for their possible effect on exercise physiology and
for understanding the different bioactivities that could be used for health
promotion (Ching et al., 2015). Physical activity modifies the balance
between oxidative stress and antioxidant defense mechanisms. For both
athletes and fitness enthusiasts, the combination of regular physical
activity and antioxidant supplement may have important restorative
effects on the body‟s oxidation reduction or redox balance (Araújo et al.,
2013).
Corn silk (Maydis stigma) is a waste material from corn cultivation,
available in abundance, and possesses antioxidant, anti-fatigue, antitumor
and anti-fungal activities (Ebrahimzadeh et al., 2009 and Hu et
al., 2010).
The seeds of pumpkin are generally considered to be agroindustrial
wastes and discarded. In some parts of the world, the seeds are
consumed raw, roasted or cooked, but only at the domestic scale. With
the discovery of their richness in protein, fibers, minerals,
polyunsaturated fatty acids and phytosterols, they are being regarded
valuable for the food industry. Also, their beneficial effects on blood
glucose level, immunity, and cholesterol, liver, prostate gland, bladder,
depression, learning disabilities and parasite inhibition are being
validated (Patel, 2013). Dietary supplementation with creatine (CrS) is
popular in the sports and fitness industry. While certain mechanisms of
action involved in improved physical exercise performance with CrS
have been established (Gama, 2011 (.
Therefore, the aim of the study was to investigate the oxidative
stress effect of corn silk, pumpkin seeds and creatine supplementation on
adult male trained rats and their exercise performance.
MATERIALS AND METHODS
Materials:
Corn silk and Pumpkin seeds were purchased from Field Crops
Research Institute, Ministry of Agriculture Giza, Egypt. Chemicals:
Creatine powder, casein, vitamin mixture, mineral mixture and cellulose
were purchased from El-Gomhoria Pharmaceutical Company, Cairo,
Egypt. Rats: fifty young adult male albino rats of Sprague- Dawely
Strain, weighting (150 ±5 g) were obtained from Animal House Colony
of Vacsera, Helwan, Egypt. Kits for blood analysis were purchased from
Gama Trade Company for Chemicals, Cairo, Egypt. The Treadmill was
purchased from Sports Equipment Store, Cairo, Egypt (YY-T900D
Luxury electronic treadmill, Motor power 3,0HP Quality in Taiwan with
231 Egypt. J. of Appl. Sci., 35 (12) 2020
an adjustable speed (0, 8 -14.8 Km/h), Running surface 1230*430mm,
and expand dimensions 1620* 700*1200 mm enabling forced exercise
training and accurate testing of fatigue in rats.
Methods:
Preparation of Dried Plants:-Fresh corn silk and pumpkin seeds
were washed by using tap water and were dried using solar energy at the
National Research Centre, Giza, Egypt in the sun at 50°C for two days
then grounded to get a fine powder .The mild temperature (45-55°C)
enable the dried product to retain its nutrients as described by (Andritsos
et al., 2003).
Active components (phenolic-flavonoids) and antioxidant activity
of corn silk and pumpkin seeds were determined chemically according to
the method described by (Brand-Williams et al., 1995)
Experimental Design:-
This study was carried out at the Animal House of Home Economic
Faculty, Helwan University. Fifty male albino rats, were housed in well
aerated cages under hygienic condition and were fed on basal diet for one
week for adaptation. During the adaptive week all rats (except negative
control group) were exposed to day after day forced treadmill running
exercise for 10 min by using treadmill according to the method of (Davis,
2009).
Rats were divided into two main groups, the first main group (n=10
rats) were fed on basal diet only and served as negative control group.
The second main group (exercised rats) were divided into 4 subgroups
(10 rats each) as followed:- Subgroup 1: were fed on basal diet and will
be run for 10, 15, 20 and 25minutes (15 days for each time period),
respectively and served as positive control group. The Subgroups (2-4)
as the same of positive control subgroup and were fed on basal diet
supplemented with 4% corn silk powder, 4% pumpkin seeds powder and
4% creatine powder, respectively.
The exhausting time protocol:
At the end of the experimental period (60 day), 5 rats of each
subgroup were exposed to the exhausting treadmill test and the run time
was recorded, then all experimental rats were scarified. Blood samples
were collected and centrifuged to obtained serum which was used for the
biochemical analysis. Liver and gastrocnemius muscle were collected
from each rat to determine glycogen content (Banchroft et al., 1996)
Biochemical analysis of serum:
Serum samples were used for the determination of glucose
described by (Trinder, 1969 and Weissman and Klein, 1958), creatine
kinase (Tietz, 1976), lactate dehydrogenase (Vassault et al., 1986),
Aspartate aminotransferase (AST) (Schumann et al., 2002), Alanine
aminotransferase (ALT) was determined in serum according to
Egypt. J. of Appl. Sci., 35 (12) 2020 232
(Sherwin, 1984)., Serum malondialdehyde (MDA) was determined by
the method of (Ohkawa et al., 1979), Serum Glutathion-s-trasferase was
determined by the method of (Habig et al., 1974).
Statistical analysis:-
The results was evaluated by statistically analyzed using
computerized program SPSS .results were expressed as mean ± SD
.differences among groups were analyzed of analysis of variance
(ANOVA) using Duncan‟s test .A p˂0.05 was considered statistically
according to (Sendecor and Cochran, 1986)
RESULTS
The total phenolic content of corn silk and pumpkin seeds are
presented in Table (1). The results indicate that corn silk contained mild
amount of myricetin acid ,benzoic acid , Salicylic acid , Neringein ,
Kampherol and mean value was 16.79, 23.84, 34.75, 61.87, 75.57 mg /
kg respectively. The Ellagic recorded the largest amount of phenolic
content in corn silk with mean value 417.10 mg / kg. On the other hand,
pumpkin seed contained mild amount of Chlorogenic, Vanillin, Caffeine,
benzoic acid, and Neringein with mean values 7.34, 8.12, 10.167, 13.47,
and 22.88 mg / kg respectively . The largest amount of phenolic contents
in pumpkin seeds was Ellagic and Catechol which recorded 71.91 and
76.84 mg/kg respectively.
Table (1): Corn silk and pumpkin seeds total phenolic content
Phenolic Compounds Corn Silk(mg / kg) Pumpkin seed(mg / kg)
Syringic acid 1.49 1.04
Vanillic acid 2.06 8.12
p- Coumaric acid 2.25 1.52
Quercetin 2.74 --
Cinnamic acid 3.63 --
Rutin 4.13 --
Chlorogenic 4.30 7.34
Ferulic acid 4.82 --
Caffeic acid 5.02 2.29
Caffeine 5.15 10.16
Myricetin acid 16.80 --
Benzoic acid 23.85 13.47
Salicylic acid 34.75 5.19
Neringein 61.88 22.88
Kampherol 75.58 1.93
Ellagic 417.10 71.91
Quinol -- 1.12
Gallic acid -- 1.72
Pyrogallol -- 1.78
Catechol -- 76.84
233 Egypt. J. of Appl. Sci., 35 (12) 2020
The data in table (2) shown the result of radical scavenging activity
for corn silk and pumpkin seeds which recorded high antioxidant activity
99.68% at 5% concentration compared to corn silk which recorded 84.07
at 5%concentration.
Table (2):The Antioxidant activity of corn silk and pumpkin seed
SN % DPPH Radical-Scavenging Activity
0.5% 1.0% 2.5% 5%
Corn Silk 20.20 41.02 60.33 84.07
Pumpkin Seed 31.79 46.53 88.50 99.68
Table (3) recorded the effect of corn silk, pumpkin seeds, and
creatine on FI, BWG, and FER in male rats which exposure to exhaustive
test according to the experimental training treadmill protocol. Results
indicate that positive control group which fed on basal diet throughout
the experiment and was trained day after day for (60) days non
significantly increased on FI level compared to (-ve) control group. Corn
silk and creatine groups significantly increased FI compared to +ve
group, otherwise pumpkin seeds group non significantly changed FI
compared to ( +ve ) group. The highest feed intake was recorded at the
creatine group. Rats in +ve control group had significant decrease
(P<0.05) in BWG compared to –ve control group. Corn silk group and
pumpkin group significantly decreased BWG compared to (+ve) control,
on the other side, creatine group significantly increased BWG compared
to (+ve) group and recorded the best group findings. Regarding to FER
of rats in +ve control group was significantly decreased compared to –ve
control group. Corn silk and pumpkin seeds significantly decreased FER
compared to +ve control group, inversely creatine group was
significantly increased FER compared to +ve control group. The best
result in FER of all tested group was creatine group.
Table (3):Effect of corn Silk, pumpkin seed and creatine on BWG, FI and
FER of rats exposure to exhaustive test
Parameters
Groups
FI g/day BWG g FER
1 (-ve) 19.68 ± 1.16c 62.25±2.99b 0.05 ± 0.001a
2 (+ve) 23.59 ± 0.54c 33.00±2.88c 0.02 ± 0.002b
3 Corn Silk 26.99 ± 1.17b -56.00±2.54e -0.03 ± 0.001d
4 pumpkin 20.00 ± 0.88c -14.50±1.58d -0.01 ± 0.001c
5 Creatine 28.03 ± 0.47a 72.75±2.21a 0.04 ± 0.001a
All value represented as mean ± SD.
Means with different superscript are significantly different (p<0.05)
Table (4) illustrated the effect of corn silk, pumpkin seeds, and
creatine on, FI, BWG, and FER in male rats which non exposure to
exhaustive test (the end of experiment) in the experimental training
Egypt. J. of Appl. Sci., 35 (12) 2020 234
treadmill protocol. Positive control group which fed on basel diet while
not exposure at the end of experiment to exhaustive treadmill test had
significant increase in FI amount compared to (-ve) group. All tested
groups had significant decrease in FI compared to +ve control group.
Rats in + ve control group had significant decrease in BWG compared to
–ve control group. Pumpkin seeds and corn silk group significantly
decreased (P<0.05) BWG compared to +ve control group, however
creatine group significantly increased compared to +ve control group.
The best findings in all test groups was creatine group. Positive control
group showed significant decreased in FER compared to –ve control
group. Pumpkin seeds and corn silk significant decreased FER compared
to +ve control group on the opposite creatine group significantly
increased compared to +ve control group and the best result from all
tested group was creatine.
Table (4): Effect of corn Silk, pumpkin seed and creatine on BWG, FI and
FER of rats non exposure to exhaustive male rats
Parameters
Groups
FI g/day BWG g FER
1 (-ve) 19.68 ± 1.16b 62.25±2.99b 0.05 ± 0.01b
2 (+ve) 22.25 ± 1.26a 24.75±2.50c 0.02 ± 0.003c
3 corn Silk 20.50 ± 2.69b -11.20±0.69d -0.01 ± 0.03d
4 Pumpkin 20.63 ± 2.18b -08.00±0.06a -0.007 ± 0.004a
5 Creatine 20.83 ± 2.34b 59.00±4.81b 0.05 ± 0.001b
All value represented as mean ± SD.
Means with different superscript are significantly different (p<0.05).
As shown in Table (5), the effect of corn silk, pumpkin seed and
creatine on AST, which illustrated significant increase in (+ve) control
group compared to (-ve) control group. Rats fed on corn silk non
significantly decrease AST compared to (+ve) control group. Whereas,
rats fed on pumpkin seeds and creatine recorded significant increase in
AST level compared to (+ve) control group and. Corn silk group marked
as the best group among all tested group. The concentration of ALT for
(+ve) control group had significant decrease compared to (-ve) control
group. Rats fed on corn silk had non-significant decrease compared to
(+ve) control group. While rats which fed on pumpkin seeds and creatine
noticed significant increase compared to (+ve) control group. Corn silk
group recorded best findings among all tested group. Results indicate that
positive control group had significant increase in ammonia level
compared with (-ve) control group. While rats which fed on corn silk and
pumpkin seeds were significantly decreased compared to (+ve) control
group. Creatine group recorded significant increase among all tested
group.
235 Egypt. J. of Appl. Sci., 35 (12) 2020
Table (5): Effect of Corn Silk, Pumpkin Seed and Creatine on liver
functions and ammonia Level on rats exposure to
exhaustive time test
Parameters
Groups
AST
u/l
ALT
u/l
Ammonia
μg /dl
1 (-ve) 122.00 ± 2.83d 41.50 ± 3.00b 73.00 ± 10.61c
2 (+ve) 128.20 ± 13.57c 34.20 ± 3.19c 104.20 ± 5.15b
3 Corn Silk 127.25 ± 9.22c 32.50 ± 7.33c 70.25 ± 4.72c
4 Pumpkin seed 148.50 ± 6.35b 64.50 ± 3.32a 51.00 ± 4.62d
5 Creatine 177.00 ± 9.90a 42.00 ± 0.81b 133.65 ± 1.30a
All value represented as mean ± SD.
Means with different superscript are significantly different (p<0.05)
Table (6) showed that rats in (+ve) control group had significant
decrease in AST level compared to (-ve) control group. Rats fed on
pumpkin seeds and creatine had significant increase in AST compared to
(+ve) group. Corn silk group recorded significant decrease compared to
(+ve) control group, and it was the best result among all tested group.
Serum ALT of (+ve) control group was significantly decreased compared
to (-ve) control group .Whereas rats fed on corn silk, pumpkin seed and
creatine significantly increased compared to (+ve) control group. The
best result for ALT at pumpkin seeds group. Results of Ammonia level of
(+ve) control group was shown significant increase compared to (-ve)
control group. Whereas corn silk group recorded nonsignificant decrease
compared to (+ve) control group. But pumpkin seeds and creatine groups
showed significant increase in ammonia level compared to (+ve) control
group.
Table (6): Effect of Corn Silk, Pumpkin Seed and Creatine on liver
functions and ammonia level on rats non exposure to
exhaustive time
Parameters
Groups
AST
u/l
ALT
u/l
Ammonia
μg /dl
1 (-ve) 122.00 ± 2.83c 41.50 ± 3.00b 73.00 ± 10.61d
2 (+ve) 110.75 ± 25.64d 33.00 ± 3.16d 93.50 ± 2.38c
3 corn Silk 99.82 ± 4.01e 42.40 ± 5.3b 91.60 ± 2.07c
4 pumpkin 137.75 ± 3.86b 36.50 ± 1.73c 125.05 ± 3.59a
5 Creatine 147.00 ± 7.81a 45.20 ± 3.96a 109.84 ± 3.40b
All value represented as mean ± SD..
Means with different superscript are significantly different (p<0.05)
Table (7) showed the effect of corn silk, pumpkin seeds, and
creatine on, glucose, CK, and LDH level in male rats which exposure to
exhaustive test. The result indicated that glucose level at (+ve) control
Egypt. J. of Appl. Sci., 35 (12) 2020 236
group was significantly increased compared to (-ve) control group. Corn
silk group and pumpkin group recorded significant decreased in glucose
level compared to (+ve) group, however creatine group recorded
significant increase in glucose compared to (+ve) control group. The best
result was recorded at corn silk group. Data of creatine kinase revealed
that (+ve) control group non-significantly decreased compared with (-ve)
control group. The finding indicated that corn silk group, pumpkin seed,
and creatine supplementation caused significant increase CK compared
with (+ve) control group, the best result among all tested groups was
creatine group. LDH level in (+ve) control group shown significant
decrease compared to (-ve) control group, but corn silk group, pumpkin
seed, and creatine recorded significant decreased compared to (-ve)
control group. The best result was corn silk group.
Table (7): Effect of Corn Silk, Pumpkin Seed and Creatine on
Glucose, CK and LDH level on rats exposure to
exhaustive time
Parameters
Groups
Glucose
mg / dl
CK
u/l
LDH
u/l
1 (-ve) 106.01 ± 5.42c 24.50 ± 5.74d 2207.75 ± 84.16a
3 (+ve) 115.02 ± 11.14b 18.20 ± 3.96d 1416.40 ± 91.50b
5 Corn Silk 96.50 ± 01.91c 676.25 ± 18.89b 787.25 ± 3.77e
6 Pumpkin seed 97.00 ± 03.83c 1185.00 ± 84.67a 965.00 ± 19.51d
9 Creatine 165.05 ± 14.14a 45.65 ± 2.36c 1301.50 ± 52.80c
All value represented as mean ± SD.
Means with different superscript are significantly different (p<0.05).
Table (8) showed the effect of corn silk, pumpkin seeds, and
creatine on, glucose, CK, and LDH level in male rats which non exposure
to exhaustive test according to the experimental training treadmill
protocol. The results indicate that (+ve) control group which fed on basal
diet and non-exposure to exhaustive time had significant increase in
glucose level compared to (-ve) control group. Corn silk and creatine
group recorded non-significant increase in glucose level compared with
(+ve) control group. However pumpkin seed group shown significant
increase compared with (+ve) control group, and also pumpkin seed
group considered as the best group among all tested group. Concerning
CK concentration in serum, the data indicated that rats in (+ve) control
group had non-significant decrease compared with (-ve) control group,
furthermore corn silk and pumpkin seed groups shown significant
increase in CK level. Creatine group was the best group among all tested
group. Results indicate that LDH level was significantly decreased in
(+ve) control group when compared with (-ve) control group but in corn
silk group, LDH was significantly decreased when compared with (+ve)
237 Egypt. J. of Appl. Sci., 35 (12) 2020
control group. The result pumpkin seeds and creatine groups indicated
significant increased values when compared with (+ve) control group.
Table (8 ): Effect of Corn Silk, Pumpkin Seed and Creatine on
Glucose, CK and LDH level of rats non-exposure to
exhaustive time
Parameters
Groups
Glucose
mg / dl
CK
u/l
LDH
u/l
1 (-ve) 106.01 ± 5.42c 24.50 ± 5.74c 2207.75 ± 84.16b
2 (+ve) 110.06 ± 11.17b 17.25 ± 0.96c 1465.50 ± 74.93d
3 corn Silk 117.60 ± 01.82b 621.60 ±106.78a 922.40 ± 31.16e
4 Pumpkin seed 158.01 ± 05.77a 50.09 ± 5.83b 2380.50 ± 91.22a
5 Creatine 124.20 ± 04.82b 20.60 ± 2.30c 2142.20 ± 168.50c
All value represented as mean ± SD.
Means with different superscript are significantly different (p<0.05).
Table (9), showed the effect of corn silk, pumpkin seeds, and
creatine on MDA and GST level in male rats which exposure to
exhaustive test according to the experimental training treadmill protocol.
Rats in (+ve) control group which fed on basal diet, recorded significant
increase in MDA concentration compared to (-ve) control group . While
rats in corn silk group, pumpkin seed group, and creatine group had
significant decrease when compared to (+ve) control group. Creatine
group consider as the best result among all tested group. The results
revealed that GST was significantly decreased in (+ve) control group
when compared to (-ve) control group, however rats fed on corn silk,
pumpkin seed and creatine shown significant increase in GST level when
compared with (+ve) control group. The best result recorded in creatine
group among all tested group.
Table (9): Effect of Corn Silk, Pumpkin Seed and Creatine on MDA
and GST level on rats exposure to exhaustive time
Parameters
Groups
MDA
nmol/ml
GST
u/l
1 (-ve) 14.40 ± 1.09b 122.58 ± 20.08b
2 (+ve) 16.13 ± 0.98a 95.60 ± 35.33d
3 Corn Silk 13.83 ± 1.00b 167.00 ± 7.86a
4 Pumpkin 13.48 ± 0.74b 111.03 ± 17.57c
5 Creatine 11.28 ± 0.84c 114.30 ± 6.65c
All value represented as mean ± SD.
Means with different superscript are significantly different (p<0.05)
Table (10) showed the effect of corn silk, pumpkin seeds, and
creatine on MAD and GST level in male rats which non-exposure to
exhaustive test. Results indicated that MDA level was decreased
Egypt. J. of Appl. Sci., 35 (12) 2020 238
significantly in (+ve) control group when compared with (-ve) control
group, while rats fed on corn silk or creatine had non-significant decrease
in MDA compared to (+ve) control group, while rats which fed on
pumpkin seed recorded a significant reduction compared to (+ve) control
group. The result revealed that GST was significantly decreased in (+ve)
control group compared to (-ve) control group. Although corn silk group,
pumpkin seed group, and creatine group caused a significant increase
compared to (+ve) control group. Creatine group was the best result
among all tested group.
Table (10): Effect of Corn Silk, Pumpkin Seed and Creatine on MDA
and GST level on rats non exposure to exhaustive time
Parameters
Groups
MDA
nmol/ml
GST
u/l
1 (-ve) 14.40 ± 1.09a 122.58 ± 10.08c
2 (+ve) 13.83 ± 1.14b 100.00 ± 14.49d
3 corn Silk 12.16 ± 1.21b 166.80 ± 13.01b
4 Pumpkin 10.18 ± 0.97c 123.56 ± 13.37c
5 Creatine 12.24 ± 0.13b 189.60 ± 13.34a
All value represented as mean ± SD.
Means with different superscript are significantly different (p<0.05).
Table (11) recorded influences of tested groups on exhaustive
treadmill time test of exercised male rats. The result indicated that rats in
corn silk group and pumpkin seed recorded 35, 45 min respectively,
whereas (+ve) control group which fed on basal diet recorded 20 minute,
these results illustrated an increase in exhaustive time test when
compared to +ve group.
Table (11): Influence of corn silk, pumpkin seeds, and creatine on
exhausting test (min) of male rats
Groups (-ve) (+Ve) Corn Silk Pumpkin seeds Creatine
Time
(min)
__ 20 35 45 25
The result in table (12) indicated that rats in (-ve) control group had
sever glycogen content in liver and was recorded the highest level
compared to all groups. The rats treated with either corn silk, pumpkin or
creatine and exposed to exhaustive treadmill test had nil glycogen
content as compared to control groups. While rats had nil, mild, sever
glycogen content at non exposure protocol and treated with creatine,
pumpkin , corn silk respectively. On the other hand, the glycogen
content in gastrocnemius muscle for the rats that exposed to an
exhaustive treadmill test was nil, mild, sever for the rats that treated with
corn silk, pumpkin, creatine respectively. While non exposure protocol
239 Egypt. J. of Appl. Sci., 35 (12) 2020
caused moderate, sever, moderate increase in glycogen content at the
groups fed on corn silk, pumpkin, creatine respectively.
Table (12) Effect of corn silk, pumpkin seeds, and
creatine on glycogen content in liver and gastrocnemius muscles
exposure and non-exposure to exhaustive treadmill test
organs groups (-ve) (+Ve )
EX
Corn
Silk
EX
Pumpkin
EX
Creatine
EX
Liver exhaustive +++ +
Non
exhaustive
+++ ++ +++ + __
Gastrocnemius
muscle
exhaustive + ++ +++ +
Non
exhaustive
+ + ++ +++ ++
+++ Sever, ++ Moderate, + Mild, - Nil
DISCUSSION
Mammalian skeletal muscle is capable of producing a large
number of free radicals and it is well recognized that a major source of
this free radical production happened during oxygen flux through the
mitochondria. Which can increase 100-fold, potentially resulting in an
increased risk of „oxidative stress‟, muscle injury and fatigue. However,
skeletal muscle is well equipped to deal with this oxidative stress by
possessing a number of antioxidant species including both endogenous
and exogenous antioxidants. It is only when there are an increased
number of free radicals, or a depletion of antioxidant levels that „redox
balance‟ is disturbed and the cell becomes vulnerable to free radical
attack (Spurway and Maclaren, 2007)
In this experimental study, albino male rats were used to examine
the oxidative stress effect of corn silk ,pumpkin seed ,and creatine after
exercised by using treadmill reached the exhaustive time test as well as ,
to determine the changes in body weight , liver enzymes and oxidative
stress biomarker.
The palatability of corn silk, pumpkin seed, and creatine powder
affected the consumption and partly accounts for the trend in the final
body weights of the animals.
Exercise of sufficient intensity and duration can lead to the
formation of reactive oxygen and nitrogen species (Fisher-Wellman and
Bloomer, 2009), which when produced in amounts that overwhelm the
antioxidant defense system may lead to a condition of “oxidative stress”.
Corn silk and pumpkin total phenolic contents which were seen in Tables
Egypt. J. of Appl. Sci., 35 (12) 2020 240
(1 and 2) may be exhibited antioxidant activity by several mechanisms
including a reduction in ischemia-reperfusion-induced oxidative stress
(Loffredo et al., 2007and Volek et al., 2002), a reduction in xanthine
oxidase activity Spierin et al., 2007)), and a free radical-scavenging
activity as shown in Table (2) (Vanella et al., 2000).
Fiber in pumpkin seeds content makes these seeds ideal for
weight loss. This dense and heavy snack can keep the body satiated for
quite a longer period of time. The fiber takes longer to digest, leaving
you full for longer and further keeps you away from bingeing on
fattening foods. Other than this, pumpkin seeds is quite a good source of
zinc, which can further help in boosting the body's metabolism. A
sluggish metabolism can hinder body's weight loss efforts to a great
extent. Hence, it is important to keep body's metabolism up and running
(Sarine, 2018). These results were consistent with the results of this
current research at tables (3 and 4).
In the line with the research results Lee et al., (2016) reported
that experimental animals that received high-fat diet along with 100
mg/kg of high maysin corn silk extract for 8 weeks showed significant
reduction of body weight compared to the high-fat diet only group.
Additionally, kidney fat and epididymal fat pad weights significant
decreased, demonstrating high maysin corn silk extract had a weightreducing
effect by decreasing fat accumulation in the body. Min et al.,
(2011) reported that administration of 100 mg/kg BW and 400 mg/kg
BW of corn silk extract significantly lowered body weight after 2 weeks
of intake in mice, which is consistent with the weight-reducing effect.
Results of the present study were in a line with Kreider et al.,
(1998) which indicate that 28 d of creatine supplementation (15.75g·d-1)
during resistance/agility training promoted significantly greater gains in
fat/bone-free mass. These findings support previous reports that creatine
supplementation may increase total body weight and/or lean body mass,
as seen at tables (3 and 4).
The most sensitive and profitable diagnostic enzymes of the liver
are aminotransferases including AST and ALT. Generally, certain
amounts of these enzymes are produced by liver cells, yet after liver
damage, the liver cells increase the secretion of the enzymes and their
plasma levels, whose increased level is a sign of liver damage (Terohid
et al., 2015). The incidence of oxidative stress in the body has other
negative effects, such as muscle fatigue, early restlessness impaired,
decreased immune function, and muscle oxidation (Bloomer and
241 Egypt. J. of Appl. Sci., 35 (12) 2020
Goldfarb, 2004). The results showed that 60 days of training protocol
had slightly alteration effect on serum levels of ALT and AST on +ve
and corn silk feeding groups exposed to exhaustive test. The lack of
alteration is inconsistent with the findings of (Kim et al., 2007 and
Ghasemnian et al., 2020). Acute strenuous exercise induces oxidative
stress and/or tissue damage in several tissues including skeletal muscle,
liver, and kidney (Suzuki et al 2003; Aoi et al., 2004 and Suzuki, 2018).
It is well known that antioxidant capacity is increased by acute or chronic
exercise (Yada et al., 2020). This research results were in a line with
above findings at tables (5 and 6) especially corn silk group showed great
improvement compared with other treated groups.
Ammonia can cross the blood–brain barrier and has the potential
to affect central neurotransmitter levels and central neural fatigue
(Hargreaves and Lawrence, 2020). An increase in ammonia
concentration occurs predominantly after high-intensity exercise, where
ammonia aids in stimulating glycolysis. With increasing fatigue, the level
of ammonia biomarker increase, reflecting the physiological status of the
athlete (Kantanista et al., 2016).These results agree with the present
study findings. As shown in the research results, corn silk play a
whiteness role in the ammonia values impairment higher than pumpkin
and creatine. The action of corn silk extract on mice was by increasing
insulin level and recovering injured beta cells, and not via increasing
glycogen or inhibiting gluconeogenesis. The results suggest that in
modern pharmacological study, corn silk extract may be used as a
hypoglycemic food or medicine for hyperglycemic people (Guo et al.,
2009). Pancreatic enzyme synthesis and secretion may change with
physical exercise (Minato, 2000). In rats, endurance running training
increases pancreatic protein content, pancreatic enzyme activity, and
basal amylase secretion (Minato, 1997), this finding was in the same line
with the present study of glucose level and agreement with (praphatsorn
et al., 2010) which found that high-intensity exercise at 75% and 90%
VO2max caused an increase of biochemical parameters in liver and
pancreas. The levels of exercise also caused histopathology changes in
the liver and pancreas.
Seeds from contain bioactive compounds which have antifatigue
activity and can elevate exercise performance. The seeds were said to
have bioactivities such as hepatoprotection, anti-diabetes, anticancer, and
anti-obesity properties (Wang, 2012). According to Bharti et al., (2013),
tocopherol isomers (α, β, γ, and δ) from raw pumpkin seeds has been
Egypt. J. of Appl. Sci., 35 (12) 2020 242
reported to be effective in the alleviation of diabetes through its
antioxidant activities. In another studies also on alloxan-induced diabetic
rats demonstrate that polysaccharide from pumpkins has hypoglycemic
effect (Xiong and Cao, 2001).This polysaccharide (50 mg/kg)
administered orally in alloxan-induced diabetic increase the body weight,
reduce water intake, and blood glucose levels of diabetic mice group
compared with diabetic control group (Perez Gutierrez, 2016).
Furthermore, the administration of fruit powder for a month in diabetic
rats significantly reduced C-reactive protein (CRP), cholesterol , glucose,
Triglycerides while insulin was increased in diabetic rats (Sedigheh et al
., 2011). Gualano et al., (2010) showed that creatine supplementation
combined with exercise alleviated glycemic conditions in T2DM.
Release of CK from muscle cells during exercise corresponds to
the degree of permeability of cell membranes and their damage resulting,
among other things, from an increase in the amount of free radicals
formed during physical activity, leading to the peroxidation of cell
membrane lipids (Banfi et al., 2012). After muscle-damaging exercise,
the enzyme CK leaks from the muscle into the circulation. It is typical for
athletes to have elevated CK during training, with reference ranges of
82–1,083 u/l in male and 47–513 u/l in female athletes suggested as
athletic norms (Mougios, 2007).
In the present study table ( 7 ) showed increasing in creatine
kinase in pumpkin seed group among others groups ,those results in the
similar line with ( AL-Zuhair et al ., 1997) which suggested that
pumpkin seeds oil was recorded ameliorated in creatine kinase levels
,while creatine group recoded lowest creatine kinase level , this finding in
the same line with (Bassit et al., 2010) observed a decrease in several
markers of muscle damage (creatine kinase, lactate dehydrogenase,
aldolase, glutamic oxaloacetic acid transaminase and glutamic pyruvic
acid transaminase) in 4 athletes after an iron man competition who
supplemented with 20 g/d plus 50 g maltodextrin during a 5 d period
prior to the competition.
During exercise, when muscles exhaust the oxygen, pyruvate gets
catalyzed into lactic acid by the lactate dehydrogenase enzyme. In
erythrocytes also pyruvate is not further metabolized due to the absence
of mitochondria but remains within the cytoplasm, finally converting to
lactate. In this reaction, NADH oxidizes to NAD+. The availability of
high intracellular concentrations of NAD is necessary to carry out the
preparatory phase of glycolysis. The net ATP production
243 Egypt. J. of Appl. Sci., 35 (12) 2020
of anaerobic glycolysis is only 2 ATP per glucose molecule as compared
to oxidative phosphorylation, which produces 36 ATP per glucose
molecule. LDH can also catalyze the dehydrogenation of 2-
hydroxybutyrate, but it is the less preferred substrate for LDH than
lactate (Adeva-Andany et al., 2014), our result in table ( 8) showing
increasing in LDH in creatine group which prove the previous
hypothesis.
Table (9 ) showing decreasing in MDA level with corn silk,
pumpkin seed, and creatine groups compare with positive control group
which reflect the great amount of antioxidant compounds like phenolic
acids, polyphenols and flavonoids scavenge free radicals such as
peroxide, hydroperoxide or lipid peroxyl and thus inhibit the oxidative
mechanisms that lead to degenerative diseases (Hu and Deng, 2011),this
met agreement with ( Bouzid et al., 2015) who found that MDA level
was higher during the recovery period in the high fitness level as
compared with the others groups,which concluded that both low and high
physical fitness levels help maintain better antioxidant defenses in older
adults. However, a higher physical fitness level, rather than a lower
physical fitness level, could increase lipid peroxidation. MDA is one the
most popular oxidative stress markers, and due to its toxicity.
Biomarkers to evaluate the antioxidant capacity include
glutathione, glutathione peroxidase, catalase, and the total antioxidant
capacity (Finsterer, 2012) GSTs are postulated as important detoxifying
enzymes that catalyse reduced glutathione-dependent reactions involved
in cellular protection against OS and toxic chemicals (Liavanera et al.,
2020), the present study GST results agree with the previous study and
(Ghahremanloo et al ., 2017) which demonstrate that Pumpkin
ameliorated oxidative stress and dyslipidemia in obese rats, leading to
decrease cardiovascular disease risk in obesity. However, some studies
showed different flavonoid synergists as most effective in
hepatoprotection. Moreover, it has been reported that corn silk effectively
increases antioxidant enzyme levels such as sodium dismutase and
glutathione peroxidase (Hu and Deng, 2011).
The data in table (11) explained the major findings of the present
study were that rats fed on pumpkin seeds and exposure to exhaustive
test recorded (45 min) the longest period among all tested groups, this
reflect Pumpkin seed and seed oil are a rich natural source of
phytosterols, proteins, polyunsaturated fatty acids, antioxidant vitamins,
carotenoids and tocopherols, and various elements, due to these
Egypt. J. of Appl. Sci., 35 (12) 2020 244
components are attributed providing many health benefits, (Perez
Gutierrez, 2016). High energy consumption during intense exercise may
cause an imbalance between the oxidation and anti-oxidation systems,
resulting in an increase in ROS and a reduction in antioxidant activities.
These behaviors lead to enhanced ROS production. Oxidative stress is
involved in both chronic fatigue and other fatigue related disorders
(Barclay and Hansel, 1991) Extreme physical stress could lead to
excessive generation of ROS in the skeletal muscle which, in turn, results
in peripheral fatigue (Allen et al., 2008).
Glycogen is an important energy material that enables movement
and provides adequate energy for muscle contraction. Energy use reduces
glycogen; meanwhile, an increase in hepatic glycogen can improve
exercise endurance (Anand, 2012)
The our histopathological examination to determine the glycogen
contents in liver and muscles variation between all rat groups which
exposure to exhaustive treadmill test indicated that rats in creatine group
were depleted glycogen content from liver but muscle stored it in sever
amount, unlike the rats in corn silk group were depleted glycogen content
from liver and muscle our result have supported from (Ivy et al., 2002)
who tested the hypothesis that a carbohydrate-protein supplement would
be more effective in the replenishment of muscle glycogen after exercise
compared with a carbohydrate supplement of equal carbohydrate content
or caloric equivalency when supplementing immediately and 2- hours
post exercise. The results indicate that the co-ingestion of protein with
carbohydrate will increase the efficiency of muscle glycogen storage
when supplementing at intervals greater than 1-hour apart, or when the
amount of carbohydrate ingested is below the threshold for maximal
glycogen synthesis. These results have important implications for athletes
who wish to limit their carbohydrate intake in an effort to control body
weight and for those athletes who participate in sports that have very
short recovery periods during competition such as basketball, ice hockey
and soccer.
Steensberg et al., (2002) suggest that pre-exercise glycogen
content may Influence glucose uptake during subsequent exercise.
However, this is only the case when delivery of substrates and hormones
remains constant. When delivery of substrates and hormones is altered,
the potential effect of glycogen on glucose uptake is negated. (Evans et
al., 2019) suggest that the type, duration, and intensity of the resistance
245 Egypt. J. of Appl. Sci., 35 (12) 2020
training program are important factors in determining the effects of
resistance training on skeletal muscle glycogen content.
On the other hand at the present study, the supplemented groups
which not exposure to exhaustive treadmill test preserves the glycogen
content either in liver or muscle, and that met agreement with hypothesis
that Decreased use of muscle causes it to become much less
metabolically efficient; unfortunately, this de adaptation becomes
apparent within a few days after cessation of exercise. Other factors
induced by endurance training include changes in cardiac output,
increases in capillary density, and increases in glycogen stores (Baynes
and Dominiczak ,2019).
Findings of these study prove that corn silk and pumpkin seeds were a
good source of flavonoid which help to scavenge the oxidative stress
substances, creatine recorded the largest glycogen quantity in
gastrocnemius muscles among supplemented groups, but it is causing an
elevation ammonia, and AST levels on the groups which exposure to
exhaustive time test, so more studies for creatine efficacy on skeletal
muscle is recommended. Moreover, it could be recommended that corn
silk and pumpkin seeds have the ability to eliminate physical fatigue and
prolong the training period without fatigue.
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تاثير حرير الذرة ,بذور اليقطين و الکرياتين عمى الاجهاد التاکسدى لفئ ا رن
الذکور المدربة
نعيم محمد ا ربح , محمد حمدى حجاج , اسماء ا زيد محمد
قسم التغذية وعموم الاطعمة – کمية الاقتصاد المنزلى –جامعة حموان – القاىرة –جميورية مصر العربية
اجريت الد ا رسة الحالية لمتحقق من مدى تاثير التدعيم بحرير الذرة ,بذور اليقطين
والکرياتين عمى الاجياد التاکسدى کمؤشر حيوى مثل وظائف الکبد ومحتوى الجميکوجين عمى
ذکور الفئ ا رن المدربة البالغة .خمسون فا آ ر من ذکور الالبينو تم تغذيتيم عمى الغذاء الاساسى
وتييئتيم لمدة اسبوع .خلال اسبوع التييئة جميع الفئ ا رن )ماعدا المجموعة الضابطة السالبة (تم
تعريضيم الى تدريب الجرى الاجبارى بالمشاية الکيربائية يوم بعد يوم لمدة 01 دقائق . تم
تقسيم الفئ ا رن الى مجموعتين رئيسيتين ,المجموعة الرئيسية الاولى )عدد 01 فئ ا رن ( تم تغذيتيم
عمى الغذاء الاساسى فقط وصنفت کمجموعة ضابطة سالبة .المجموعة الرئيسية الثانية )الفئ ا رن
المدربة(حيث قسمت الى 4 مجموعات فرعية ) 01 فئ ا رن فى کل مجموعة ( کالتاالى:-
المجموعة الفرعية 1:-تم تغذيتيم عمى الغذاء الاساسى وتم الجرى لفترة 52,51,02,01 دقيقة
عمى الت والى ) 02 يوم لکل فترة زمنية ( وصنفت کمجموعة ضابطة موجبة. المجموعة الفرعية
4-5 (:-مثل المجموعة الفرعية الموجبة وتغذت عمى الغذاء الاساسى مدعم ب 4%مسجوق (
حرير الذرة , 4%مسحوق بذور اليقطين , 4%مسحوق الکرياتين عمى التوالى .فى نياية فترة
التجربة ) 01 يوم( 2 فئ ا رن من کل مجموعة فرعية تم تعريضيم الى اختبار الاجياد المستنفذ
بالمشاية الکيربائية وتم تسجيل وقت الجرى ,بعد ذلک تم ذبح کل فئ ا رن التجربة .برىنت نتائج
ىذه الد ا رسة عمى ان حرير الذرة وبذور اليقطين مصدر جيد لمفلافونات والذى يساعد فى ازلة
نواتج الاجياد التاکسدى وتحسين وظائف الکبد, عمى الرغم ان سجل الکرياتين الکمية الاکبر من
محتوى جميکوجين عضمة الساق بين کل المجموعات المدعمة ولکنو سبب ارتفاع فى مستويات
فى المجموعة التى تعرضت الى اختبار وقت الاجياد المستنفذ . يمکن AST الامونيا و
التوصية بحرير الذرة وبذور اليقطين لقدرتو عمى تقميل الارىاق البدنى واطالة فترة التتمرين دون
حدوث ارىاق.
الکممات المفتاحية:- حرير الذرة , بذور اليقطين, مکملات الکرياتين ,کرياتين کاينيز , لاکتات
دى ىيدروجينيز ,مشاية کيربائية ,الاجياد التاکسدى .
Egypt. J. of Appl. Sci., 35 (12) 2020 252
