EFFECT OF MUSHROOM FUNGUS FEEDING ON INDUCED HEPATOTOXICITY RATS

EFFECT OF MUSHROOM FUNGUS FEEDING ON
INDUCED HEPATOTOXICITY RATS
Aya A.Shehata1, Ahmed A.Farrag1, Eman E.Elhady2
and Hussein H. Elsheikh3
1Nutrition and Food Science Department, Faculty of Home Economics, Helwan
University, Cairo, Egyp
2Nutrition and Food Science Department, Faculty of Home Economics, Al-Azhar
University, Tanta, Egypt
3Department of Botany and Microbiology, Faculty of Science, Al-Azhar University,
Cairo, Egypt.
Key Words: Mushroom (Agaricus bisporus), Extract, Hepatotoxicity, Rats.
ABSTRACT
In last decades, Mushroom is used widely in feeding for its
medicinal properties, many studies confirmed its role as antioxidant,
anticancer, antidiabetic, antiallergic, immunomodulating, cardiovascular
protector, anticholesterolemic, antiviral, antibacterial, antiparasitic,
antifungal, detoxification, and hepatoprotective effects; they also protect
against tumor development and inflammatory processes. The aim of the
present study was to investigate the effect of mushroom (Agaricus
bisporus) as a part of diet on hepatotoxic rats. Thirty adult male albino
rats (Sprague-Dawley strain), weighing about (200±10g) were divided
randomly into two main groups as follow: the first group (-ve control= 6
rats) was fed on basal diet. The second group (24 rats) were fed on basal
diet and injected with CCl4, using a five necrogenic dose to induce acute
liver damage, then divided into 4 groups from group 2 to group 5. Group
2 (+ve control) fed on basal diet. Group 3 and 4 fed on basal diet
supplemented with 20% of dry uncooked mushroom and 20% of dry
cooked mushroom, respectively. Finally, group 5 fed on basal diet
supplemented with10% of duple (aquatic and ethanolic) extract of
mushroom. At the end of the experimental period (4 weeks), rats were
scarified and serum was collected for biochemical analyses. Results
indicated that serum concentrations of aspartate aminotransferase (AST),
alanine aminotransferase (ALT), alkaline phosphatase (ALP), total
bilirubin and malondialdehyide (MDA) level was significantly increased
(P< 0.05) in the positive control group compared with the negative
control and significantly (P<0.05) decreased in serum of Reduced
Glutathione (GSH), superoxide dismutase (SOD) and catalyzes (CAT). It
also indicated that supplemented diet with dry uncooked, dry cooked and
its extract reversed these changes that caused by CCl4 administration.
Histological examinations of studied rat's livers sustained biochemical
and enzymatic results. It could be recommended that Mushroom is
worthy treating on Hepatotoxicity.
Egypt. J. of Appl. Sci., 35 (11) 2020 127-142
INTRODUCTION
The liver is the largest solid organ, the largest gland and one of the most
vital organs that functions as a centre for metabolism of nutrients and
excretion of waste metabolites (Ozougwu and Eyo, 2014). A total loss of
liver function could leads to death within minutes, demonstrating the liver„s
great importance (Ozougwu, 2014).
The liver is a reddish-brown wedge-shaped organ with four lobes of
unequal size and shape. A human liver normally weighs 1.44–1.66 kg, and
has a width of about 15 cm .It is both the heaviest internal organ and the
largest gland in the human body (Cotran et al., 2005).
The liver has a wide range of functions, including detoxification of
various metabolites, protein synthesis, and the production of biochemicals
necessary for digestion. It also plays a role in metabolism, regulation of
glycogen storage, decomposition of red blood cells and hormone production.
The liver is an accessory digestive gland and produces bile, an alkaline
compound which aids in digestion via the emulsification of lipids (Tortora
et al., 2008).
Treatment options for common liver diseases are limited, and therapy
with modern medicine may lack in efficacy. So, there is a need for effective
therapeutic agents with a low incidence of side effects. The natural
antioxidants, more recently, have attracted considerable attention of users
and researchers largely on account of adverse toxicological reports on some
synthetic antioxidants and growing awareness among consumers
(Ramalakshmi et al., 2007).
In fact, mushrooms may have diversity of chemicals ranging from
bitter compounds that stimulate digestive system, phenolic compounds for
antioxidant and many other pharmacological properties, including
antibacterial and antifungal, tannins that work as natural antibiotics, diuretic
substances, and alkaloids. The usage of natural drugs for the treatment of
liver diseases has increased all over the world. Developing therapeutically
effective agents from mushrooms as natural source may reduce the risk of
toxicity when the drug is used clinically (Girish and Pradhan, 2008;
Chang and Wasser, 2012 and Finimundy et al., 2013).
Mushrooms have been considered as ingredient of gourmet cuisine
across the globe; especially for their unique favor and have been valued by
humankind as a culinary wonder. More than 2,000 species of mushrooms
exist in nature, but around 25 are widely accepted as food and few are
commercially cultivated. Mushrooms are considered as a delicacy with high
nutritional and functional value, and they are also accepted as nutraceutical
foods; they are of considerable interest because of their organoleptic merit,
medicinal properties, and economic significance (Chang and Miles, 2008
and Ergo¨nu¨l et al., 2013). However, there is not an easy distinction
between edible and medical mushrooms because many of the common
128 Egypt. J. of Appl. Sci., 35 (11) 2020
edible species have therapeutic properties and several used for medical
purposes are also edible (Guillamon, 2010).
The most cultivated mushroom worldwide is Agaricus bisporus,
followed by Lentinus edodes, Pleurotus spp. and Flammulina velutipes.
Mushrooms production continuously increases, China being the biggest
producer around the world (Aida et al., 2009 and Patel and Goyal, 2012).
Mushrooms could be an alternative source of primary and secondary
metabolites that necessary for human nutrition as vitamins, peptides and
proteins (Alves et al., 2012). Mushrooms have a great nutritional value since
they are quite rich in protein, with an important content of essential amino
acids and fibers, poor fat but with excellent important fatty acids content.
Moreover, edible mushrooms provide a nutritionally significant content of
vitamins (B1, B2, B12, C, D, and E) (Mattila et al., 2001 and Heleno et al.,
2010). Thus, they could be an excellent source of many different
nutraceuticals and might be used directly in human diet and to promote
health for the synergistic effects of all the bioactive compounds present
(Ferreira et al., 2010 and Vaz et al., 2010).
MATERIALS AND METHODS
Materials:
Mushroom (Agaricus bisporus) was obtained from Ministry of
agricultural in giza. Carbon tetrachloride CCl4 was obtained from Sigma-
Aldrich, Germany. The contents of the basal diet; casein, all vitamins,
minerals, cellulose, choline and starch were obtained from El-Gomhoria
Company, Cairo, Egypt. Kits for biochemical analysis required for
estimating parameters used in this study were purchased from the Gamma
Trade Company for Pharmaceutical and Chemicals, Dokki, Egypt.
Experimental animals: Adult male Sprague-Dawley rats (n =30) which
weighing (200+10g) were purchased from Farm of experimental animals in
Giza, Egypt.
Methods:
1. Preparation of mushroom samples
The fungal materials (15) kg of healthy apparatus fruiting bodies of
Agaricus bisporus mushroom) were brought to the laboratory in sterile bags
and processed within a few hours after. The fungal materials were rinsed
gently in running water to remove dust and debris. After proper washing,
stems and fruiting bodies samples were cut into small pieces and dried with
air. The dry mushroom, then divided to three parts: the first was dry, the
second was dry and cooked and the later was extracted with aqueous and
ethyl alcohol.
2. Induction of Hepatotoxicity
Hepatotoxicity was induced using the method describe by Pawa and
Ali ,(2004), were employed carbon tetrachloride (CCl4) to induce hepatic
toxicity in rats, using a five necrogenic dose (1.5 ml/kg body weight of 80%
Egypt. J. of Appl. Sci., 35 (11) 2020 129
CCl4 in corn oil) which was equivalent to 1/5 of the oral LD50 in mice
(Abou Gabal et al., 2007) for one week After this period, a blood samples
were taken from injected rats for measuring serum aspartate
aminotransferase (AST), alanine aminotransferase (ALT) , alkaline
phosphatase (ALP) and total bilirubin concentration to be sure that all rats
have been suffering from hepatotoxicity.
2. Preparation of Basal Diet and Experimental Animal Design:
The basal diet was formulated according to AIN-93M diet (Reeves et
al., 1993).
Animals (30 rats) were divided into five groups and housed in well
aerated cages under hygienic conditions of humidity, temperature (20-25°C)
and light (12-h light: 12-h dark cycle) and fed on basal diet for one week
before starting the experiment for acclimatization in animal biological
studies Lab of RCMB - Al Azhar University. They were left for seven days
as adaptation period and they were allowed to feed standard laboratory food
and water.
The first main group (G1, 6 rats) was fed on the basal diet during the
experimental period as a negative control group. The rest of the animals
(n=24) were induce hepatic toxicity in them and divided into 4 groups (G2
to G5, 6 rats in each) as follows: Group (2): six rats with hepatotoxicity were
fed on basal diet only as positive control group, Group (3): six rats with
hepatotoxicity were fed on basal diet supplemented with 20% of dry
uncooked mushroom, Group (4): six rats with hepatotoxicity were fed on
basal diet supplemented with 20% of dry cooked mushroom and Group (5):
six rats with hepatotoxicity were fed on basal diet supplemented with 10%
of duple (aquatic and ethanolic) extract of mushroom.
At the end of the experimental period (4weeks), rats were sacrificed
after overnight fasting. Blood samples were immediately collected in clean
and dry tubes from the portal vein and left to clot at room temperature.
Blood samples were centrifuged at 3000 rpm for 15 minutes to separate
serum. Serum was carefully separated into dry clean tubes and allowed to be
frozen at -20 c until the determination of the tested parameters.
3. Biological Parameters: feed intake (FI), body weight gain % (BWG%)
and feed efficiency ratio (FER) were measured using the following
equations as described by Chapman et al., (1959); BWG % = (Final body
weight – Initial body weight) / Initial body weight X 100
FER = Body weight Gain (g)/ Food consumed (g).
Chemical analysis of serum:
Serum aspartate aminotransferase (AST) was measured using
Spectrophotometer at 505 nm according to themethod described by Young,
(2001), and Serum alkaline phosphatase (ALP) was determined according to
Roy, (1970). Serum total bilirubin concentration was determined according
to Young, (2001). Serum total cholesterol was determined according to the
130 Egypt. J. of Appl. Sci., 35 (11) 2020
method described by Allain et al., (1974). Serum cholesterol was
determined according to the method described by Fossati and principel,
(1982). Serum HDL-C was determined according to Albers et al., (1983).
Concentration of VLDL-c and LDL-c were estimated according to the
method described by Friedewald et al., (1972).
Determination of Oxidative stress and Antioxidant Biomarkers:
The animals were sacrificed at the end of treatment and the liver was
perfused and excised. The liver portion was rinsed in ice cold normal saline,
followed by 0.15 M Tris-HCl (pH 7.4) blotted dry and weighed. A 10 % w/v
of homogenate was prepared in 0.15 M Tris-HCl buffer and processed for
the estimation of lipid peroxidation (MDA). A part of homogenate after
precipitating proteins with Trichloroacetic acid was used for estimation of
glutathione. The rest of the homogenate was centrifuged at 1500 rpm for 15
min at 4°C. The supernatant thus obtained was used for the estimation of
SOD, and CAT activity.
Reduced Glutathione (GSH) was determined according to Ellman,
(1959); Super Oxide dismutase (SOD) was determined according to
Kakkar et al., (1984); catalase (CAT) was determined according to Aebi,
(1974) and malondialdehyde (MDA) determined according to Mansour et
al.,(2016).
Histopathological examination:-
The liver of the scarified rats were taken and immersed in 10%
formalin solution. The fixed specimens were then trimmed, washed and
dehydrated in ascending grades of alcohol. Specimens were then cleared in
xylol, embedded in paraffin, sectioned at 4-6 microns thickness, stained with
heamtoxylin and eosin stain for histopathological examination as described
by Carleton, (1979), then observed under a light microscope (Olympus,
Japan).
Statistical analysis:
Results of biochemical analysis and biological evaluation of each
group were statistically analyzed as mean ± SD using one way ANOVA as
described by SAS, (2006).
RESULTS AND DISCUSSION
Among the antioxidant compounds in mushrooms, both
polysaccharides and phenolic compounds have attracted much attention. In
many studies comparing antioxidant activities in alcoholic extracts from
different mushrooms, positive correlations were found with the total
phenolic content (Savoie et al., 2008; Jiang et al., 2010 and Liu and
Wang, 2012).
The detoxification effects of mushroom and mushroom extract on
CCl4 induced hepatic injury in rats are shown through the following results.
The results on Table (1) showed the effect of supplemented with dry
uncooked mushroom; dry cooked mushroom and duple (aquatic and
Egypt. J. of Appl. Sci., 35 (11) 2020 131
ethanolic) extract of mushroom on feed intake (FI), body weight gain %
(BWG%) and feed efficiency ratio (FER) of hepatotoxic rats. Feed intake
was increased in the negative control group, compared to the positive
control group. While treated groups were close to negative control group.
Table (1): Effect of Mushroom and Mushroom Extract on Feed
Intake (FI), Body Weight Gain (BWG) and Feed
Efficiency Ratio (FER) of Hepatotoxic Rats:
Parameters
Groups
Biological evaluations
FI(g/d) BWG % FER
G1:-ve control 17.8 12.32±1.64a 0.68±0.09a
G2:+ve control 14.70 2.08±0.62d 0.14±0.04d
G3:20%uncooked mushroom 18.9 5.31±0.50c 0.27±0.03c
G4:20% cooked mushroom 18.6 10.03±1.19b 0.054±0.10b
G5: 10% of duple extract of mushroom 18.2 13.86±1.39a 0.75±0.07a
*Mean values are expressed as means ± SD.
*Mean values at the same column with the same superscript letters are not
statistically significant at P<0.05.
Adding mushroom on rat's diet was bustle in the taste. Thus, its impact
on the total weight as a positive effect and these results were striking with
previous studies on other materials and extracts (Nwanjo and Orjiako,
2006; Chen et al., 2008 and Wasser, 2011).
As seen in Table (2) serum concentrations of aspartate
aminotransferase (AST), alanine aminotransferase (ALT), alkaline
phosphatase (ALP) and total bilirubin of hepatotoxic rats were significantly
increased (P<0.05) elevated by CCl4 administration (positive control group)
compared with negative. It was observed significant (P<0.05) reduce in
serum ALT,AST,ALP and total bilirubin levels for all treated groups with
mushroom and the duple (aquatic and ethanolic) extract of mushroom
compared to the positive control group. The highest improvement for liver
functions was observed at the group that fed on 10% of duple (aquatic and
ethanolic) extract of mushroom.
The hepatoprotective effects of mushroom (Macrocybe gigantea)
ethanolic extract on CCl4 induced hepatic injury in mice are recorded by
Acharya et al., (2012). The CCl4 receiving group as expected, revealed
significantly higher increase in liver function indices such as SGPT, SGOT
and ALP (P<0.05) compared to the normal group. Treatment with ethanolic
extract significantly (P<0.05) lowered the activities of serum marker
enzymes, bilirubin, comparable to standard drug silymarin and towards
normalization. Serum transaminases (SGPT and SGOT) were inhibited by
55.02% and 62.59% respectively compared with the control group animals
whereas the extract showed inhibition of 39.26% in ALP level with respect
to the control set. The increased activity of serum enzymes may explain cell
membrane break down and death (Kaplowitz et al., 1986). CCl4 intoxication
132 Egypt. J. of Appl. Sci., 35 (11) 2020
even produced a significant (P<0.05) rise in serum bilirubin thereby
indicating hepatic damage (Plaa and Hewitt, 1982).
Table (2): Effect of Mushroom and its Extract on serum
concentrations of aspartate aminotransferase (AST),
alanine aminotransferase (ALT), alkaline phosphatase
(ALP) and total bilirubin of hepatotoxic rats.
Parameters
Groups
Liver enzymes
AST
(U/L)
ALT
(U/L)
ALP
(U/L)
Total Bilirubin
(Mg/dl)
G1:-ve control 81.16±2.85e 34.17±2.31b 80.84±0.34d 0.19±0.04c
G2:+ve control 450.83±8.6a 145.17±7.75a 140.66±0.21a 0.53±0.08a
G3:20%uncooked
mushroom
220.66±5.89b 38.50±1.87b 111.29±0.47b 0.31±0.10b
G4:20% cooked
mushroom
173.6±8.91c 33.67±2.06b 107.97±0.50bc 0.22±0.05c
G5: 10% of duple
extract of mushroom
161.33 ±5.60 d 19.42±2.24c 104.35±0.25c 0.19±0.05c
*Mean values are expressed as means ± SD.
*Mean values at the same column with the same superscript letters are not
statistically significant at P<0.05.
The enhancement of results on liver functions may be due to that
mushroom contains Phenol, flavonoid, ascorbic acid and carotenoids
concentrations contained in the five Agaricus sp. mushroom extracts.
Phenols were the major antioxidant components found in the Agaricus
extracts (Andrera et al., 2009). Ergosterol, a phenolic compound extracted
from white button mushroom (Agaricus bisporus) showed inhibitory effect
on cancer cell line in vitro by aromatase inhibition without side effect
(Lillian et al., 2002).
The hepatoprotctive property of mushroom extract may also be
because it other properties like anti-inflammatory. (Chang and Schiano,
2007; Barros et al., 2008 and Maares and Haase, 2016) reported that
phenolic and antioxidant properties of (Agaricus bisporus) caused the
substantial attenuation of cell inflammation.
Data in Table (3) also revealed that; TC, TG, LDL-C and VLDL-C
were significantly (P< 0.05) increased in the positive control group
compared with the negative control group. Results showed that all groups
that were treated with mushroom significantly decreased (P< 0.05) in serum
TC, TG and VLDL-C compared to the positive control group.
Serum LDL-C level, results showed that all groups treated with
mushroom significantly decreased (P< 0.05) compared to the positive
control group. Rats were fed on 10% of duple (aquatic and ethanolic) extract
of Mushroom had the best result for reducing serum LDL-C level.
Regarding serum HDL-C level, results demonstrated a significant (P< 0.05)
decrease in serum HDL-C level of the positive control group compared to
the negative control group. Also, it was observed that all treated groups with
Egypt. J. of Appl. Sci., 35 (11) 2020 133
mushroom significantly increased (P< 0.05) compared to the positive
control group. Rats with fed on Rats were fed on 10% of duple (aquatic and
ethanolic) extract of mushroom considered the best result for increasing
serum HDL-C level. The highest improvement for lipid profile was
observed at the group that fed on 10 % of duple (aquatic and ethanolic)
extract of mushroom. Previous studies highlights on the Flavonoids and
polyphenols as active metabolites in mushroom and playing important role
as antioxidant and antilipidemic agents (Shieh et al., 2001; Wang et
al., 2002; Lakshmi et al ., 2006 Hu et al., 2006; Du et al., 2015 and
Alshammari et al., 2017).
Table (3): Effect of mushroom and its extract on serum
concentrations of total cholesterol (TC), triglyceride
(TG), high density Lipoprotein cholesterol (HDL-C),
low density Lipoprotein cholesterol (LDL-C) and Very
Low density Lipoprotein cholesterol (VLDL-C) of
Hepatotoxic Rats.
Parameters
Groups
Lipid Profile
TC TG HDL-C LDL-C VLDL-C
mg/dl
G1: -ve control 85.40±1.89a 62.05±3.41a 60.00±2.55a 20.07±1.40b 10.83±1.26b
G2: +ve control 92.67±2.61b 97.95±2.94d 43.60±3.72c 23.62±3.21a 21.66±1.26b
G3:20%uncooked
mushroom
78.33±3.47d 90.00±3.47b 49.33±1.40d 10.65±0.89d 17.86±1.56a
G4:20% cooked
mushroom
81.80±2.28c 92.97±2.59a 48.78±1.98d 17.52±2.004c 18.72±1.56 a
G5: 10% of duple
extract of
mushroom
73.55±1.69d 70.53±3.63c 56.55±2.09b 8.58±1.10d 12.10±1.58b
*Mean values are expressed as means ± SD.
*Mean values at the same column with the same superscript letters are not
statistically significant at P<0.05.
Results Table (4) also, showed that serum GSH, SOD, and CAT was
significantly decreased (P< 0.05) in the positive control group compared
with the negative control group. It was clear that, there was significant (P<
0.05) increase in serum GSH, SOD and CAT for all treated groups with
mushroom compared to the positive control group. Rats were fed on 10% of
duple (aquatic and ethanolic) extract of mushroom considered the best group
for increasing serum GSH, SOD and CAT. Results also, showed that serum
MDA level was significantly increased (P< 0.05) in the positive control
group compared with the negative control group. Moreover, all treated
groups with mushroom were significantly (P< 0.05) decreased in serum
MDA level compared with the positive control group. The best group that
reduced serum MDA level was the group that treated with 10% of duple
(aquatic and ethanolic) extract of mushroom.
134 Egypt. J. of Appl. Sci., 35 (11) 2020
Table (4): Effect of Mushroom and its Extract on Reduced
Glutathione (GSH), superoxide dismutase (SOD),
catalyzes (CAT) and malondialdehyide (MDA) of
Hepatotoxic Rats:
Parameters
Groups
Oxidative enzymes
Catalase SOD GSH MDA
(U/mg) (μmol/dL)
G1:-ve control 6.07±0.29a 15.75±0.7a 38.32±1.20a 10.49±0.23d
G2:+ve control 3.09±0.68c 8.66±0.62c 17.01±0.79d 22.18±0.44a
G3:20%uncooked
mushroom
4.67±0.27b 13.64±0.8b 33.78±1.1c 19.41±0.19b
G4:20% cooked
mushroom
4.38±0.44b 15.56±0.32a 34.4 ±1.42c 17.02±0.31c
G5: 10% of duple extract
of mushroom
4.80±0.31b 15.94±0.61a 35.90 ±0.71b 19.52±0.24b
*Mean values are expressed as means ± SD.
*Mean values at the same column with the same superscript letters are not
statistically significant at P<0.05.
The primary function of antioxidant compounds in fungi is to prevent
cell damage induced by ROS. Under normal conditions, ROS are cleared
from the cell by action of superoxide dismutase (SOD). SOD catalyzes the
conversion of O2 to H2O2 which is then decomposed in the presence of
catalase (CAT) into water and oxygen. In addition, glutathione (GSH) and
glutathione-related enzymes also play an important role against ROS, which
are reduced by GSH in the presence of glutathione peroxidase (GPx) and
GSH is regenerated by glutathione reductase (GR). The oxidative stress can
be removed by the induction of these antioxidant enzymes. In a few in vivo
studies, generally an aqueous extract of A. subrufescens was administered
orally to rats or mice. During aging of rats, de Sa-Nakanski et al., (2014)
showed that A. subrufescens was protective mainly to the brain against the
oxidative stress by increasing activity of antioxidant enzymes such as SOD
and CAT. An improvement in the functionality of mitochondria from brain
as evidenced by an increase in the activity of respiratory chain enzymes was
also observed. Polysaccharides appear to be involved in this activity of
mushrooms related to the antioxidant enzymes, as has been shown for some
fungi such as L. edodes, Ganoderma sp, Auricularia sp, Grifola frondosa,
Hericium erinaceus and Pleurotus abalones (Huang and Nie, 2015). Many
other edible mushrooms were reported to have in vitro and in vivo
antioxidant properties due to the presence of various putative bioactive
compounds such as polysaccharides, vitamins, carotenoids, micronutrients,
and polyphenols (Kozarski et al., 2015). SOD, CAT, and GSH-dependent
and recycling enzymes are also present in mushroom cells and they
contribute to their antioxidant and detoxicant defences.
Egypt. J. of Appl. Sci., 35 (11) 2020 135
The histopathological studies was performed to provide direct
evidence of the possibility of the mushroom being able to minimize
disruption of structure of hepatocytes and accelerates hepatic regeneration
thus decreasing the leakage of SGPT, SGOT and ALP into the circulation.
Histopathological examination:-
Histopathological analysis of Rats liver sections using H&E staining
(×200) are showed in photo (1); A: Section from a liver of negative control
rat (fed on basal diet) revealed that no marked histopathological changes.,B:
The liver section of positive control rats (fed on basal die and injected with
CCl4( showed that dilatation of hepatic sinusoids and proliferation of oval
cells are considered indication of hepatic carcinoma. ,C: Liver Section of
rats fed on 20% of dry uncooked mushroom showed a moderate
improvement in histological structure of liver and few vacuolar degeneration
of hepatocyte ,D: Liver Section of rats fed on 20% of dry cooked
mushroom indicated a moderate improvement in histological structure of
liver with mild dilatation sinusoids of capillaries and mild proliferation fiber
connective tissue in portal area and E: Liver tissue sections prepared from
rats fed on 10% of duple (aquatic and ethanolic) extract of mushroom
marked improvement in histological structure of liver and nearly disappear
of oval cells administration groups exhibited less cavitation and necrosis
compared to these shown in B. group.
Results of histopathological examinations of liver were supported by
Wu et al., (2011) who indicated that CCl4-treated rats caused a severe
hepatocellular degeneration, necrosis, and congestion of the sinusoids, along
with periportal mononuclear cell infiltration due to toxicity.
In the other hand, Standish et al., (2006), reported that mice in the
negative control group exhibited normal, well-defined histological
structures, without any signs of vascular or inflammatory changes; no
cavitations, necrosis or fibrosis were found in normal control sections. The
histopathological analysis of the liver revealed signs of toxicity after
administration of CCl4. This toxicity was significant in comparison with the
negative group and included cavitations, fibrosis in broad areas, mild
vascular congestion and moderate inflammatory changes with congested
sinusoids, nuclear changes, and centrilobular necrosis. The broad cavitations
and fibrosis in livers were somewhat attenuated in mice treated with low or
high doses of Agaricus blazei Murrill (ABM) during the experimental
periods. ABM administration for mice did result in fewer cavitations and less
fibrosis in the liver. ABM treatment also elevated the survival rate of mice
after liver injury induced by CCl4. ABM reduced apparent liver injury caused
by CCl4 for mice by histopathological assessment in a dose-dependent
manner.
136 Egypt. J. of Appl. Sci., 35 (11) 2020
Photo (1): A,B,C,D and E Histopathological analysis of Rats liver
sections using H&E staining (×200). A: Negative control
rat (fed on basal diet).,B: Positive control rats (fed on basal
die and injected with CCl4( ,C: Uncooked mushroom ,D:
Cooked mushroom and E: Duple (aquatic and ethanolic)
extract of mushroom.
A B
C D
E
Egypt. J. of Appl. Sci., 35 (11) 2020 137
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تأثير التغذية بفطر عيش الغ ا رب عمي الفئ ا رن المصابة بالتسمم الکبدي
آية عاطف شحاته*- أحمد عوض ف ا رج*- إيمان السيد عبد الهادى**- حسين حسنى الشيخ***
*قسم التغذية وعموم الأطعمة - کمية الاقتصاد المنزلي - جامعة حموان- القاهرة
** قسم التغذية وعموم الأطعمة - کمية الاقتصاد المنزلي – جامعة الأزهر- طنطا
*** قسم النبات والمکروبيولوجى - کمية العموم بنين - جامعة الأزهر- القاهرة
فٙ انؼق دٕ الأخٛشح, رى اعزخذاو فطش ػٛش انغشاة ػهٙ طَبق أعغ فٙ انزغزٚخ ظَشا
نخظبئظ انطجٛخ ان إعؼخ. أکذد انؼذٚذ ي انذساعبد انخ إص ان ًًٛضح ن ک ؼًبد نلإنز بٓثبد ,
خبفغ نغکش انذو , يؼبد نهجکزٛشٚب ئؼبد ن شُبؽ الأ سٔاو. ر ذً سٕد انذساعخ ف زْا انجذش
د لٕ يؼشفخ رأصٛش فطش ػٛش انغشاة ي عَٕ الأجبسکظ ثٛغج سٕط ػهٙ يشػ انزغ ىً انکجذ ± فٙ انفئشا .ٌ أجشٚذ انذساعخ ػهٙ صلاص فأسا ي عَٕ الأنجٛ ,ُٕ رزشا حٔ أ صٔا ىَٓ ي ) 200
رى )G 10 جى( دٛش رى رقغٛ ىًٓ إنٙ يج ػًٕزٛ أعبعٛزٛ : ان جً ػًٕخ الأ نٔٙ ) 6 فئشا 1
رغزٚز ىٓ ػهٙ انغزاء الأعبعٙ ؽ إل فزشح انزجشثخ رٔ ضًم ان جً ػًٕخ انؼبثطخ انغبنجخ . ان جً ػًٕخ
انضب َٛخ ) 24 فأسا( رى رغزٚز ىٓ ػهٙ انغزاء الأعبعٙ دق ىُٓ رذذ انجهذ ث بًدح ساثغ که سٕٚذ انکشث ف ط سٕح خ ظً جشػبد يزغب ٚٔخ ) 1.5 يم / کغى ي صٔ انجغى ي 00 ٪ ي )CCl4(
فٙ صٚذ انزسح( يب ٚؼبدل أدذ ي خ غًخ أجضاء ي انجشػخ ان ظُف ي هٓکخ )CCl4(
نهفئشا انجشػخ انکبفٛخ لإدذاس ع ًٛخ انکجذ ث ىٓ. صى رقغى ان جً ػًٕخ انضب َٛخ إنٙ )LD50(
ش هًذ کم ي بُٓ 6 فئشا يظبثخ ثبنزغ ىً )G5 –G أسثغ يج ػًٕبد يزغب ٚٔخ فشػٛخ يزغب ٚٔخ ) 2
انکجذ کبنزبن : ان جً ػًٕخ انفشػٛخ ) 2(, رى رغزٚز ىٓ ػهٙ انغزاء الأعبعٙ فقؾ رٔؼزجش يج ػًٕخ
ػبثطخ ي جٕجخ. ان جً ػًٕخ انفشػٛخ ) 3(, رى رغزٚز ىٓ ػهٙ انغزاء الأعبعٙ ان ذًز ػهٙ ٪20 ي فطش ػٛش انغشاة انجبف غٛش ان طًج رٕ. ان جً ػًٕخ انفشػٛخ ) 4(, رى رغزٚز ىٓ ػهٙ انغزاء
الأعبعٙ ان ذًز ػهٙ ٪20 ي انفطش ان طًج رٕ انجبف نکم کجى ي انغزاء الأعبعٙ.
ان جً ػًٕخ انفشػٛخ ) 5(, رى رغزٚز ىٓ ػهٙ انغزاء الأعبعٙ ان ذًز ػهٙ ٪10 ي يغزخهض
ص بُئٙ )يبء کذ لٕ( ي انفطش. اعز شًد انزجشثخ ن ذًح أسثؼخ أعبثٛغ, فٔٙ بَٓٚخ فزشح انزجشثخ, رى
رششٚخ انفئشا أنذظ لٕ ػهٙ ديبء بْ لإجشاء انزذبنٛم انجٛ کٕٛ ًٛبئٛخ إٔجشاء انفذ صٕ
انزششٚذٛخ. ظَٔشا ن بً ٚذز ٕٚ فطش ان شًش ؤ ي يشکجبد ن بٓ شَبؽ يؼبد نلأکغذح, فقذ نؼجذ زْ ان إًد د سٔا بْيب ک ؼًبداد نهزغ ىً انکجذ ف انفئشا ان شًٚؼخ انز اشز هًذ رغزٚز بٓ ػه انفطش
ف ط سٕ ان خًزهفخ, دٛش أظ شٓد زَبئج رذهٛم يغز ٕٚبد ا ضَٚ بًد انکجذ أنظفشاء انکهٛخ ف ف ان غُٛج انکجذ نهفئشا کب ذَ يشرفؼخ MDA ديبء انفئشا کٔزنک الإ ضَٚى ان ؤًکغذ ي عَٕ
ثشکم يهذ ظٕ فٙ ان جً ػًٕخ انؼبثطخ ان جًٕجخ يقبس خَ ثبن جً ػًٕخ انؼبثطخ انغبنجخ, ػلا حٔ ػهٙ
کٔزنک SOD – GSH رنک دذ سٔ ا خَفبع يهذ ظٕ فٙ يغز إلإ ضَٚ بًد ان ؤًکغذح ي عَٕ
ک بً أظ شٓد ان زُبئج ثق حٕ أ إػبفخ فطش ػٛش انغشاة ئغزخهظ إن غزاء انفئشا .CAT
ان شًٚؼخ ػکظ زْ انزغٛٛشاد انزٙ أدذص بٓ ساثغ که سٕٚذ انکشث .ٌٕ ک بً أ انزغزٚخ ثفطش ػٛش
انغشاة خففذ ي زْ اٜصبس انؼبسح نشاثغ که سٕٚذ انکشث ػه الأ غَجخ انکجذٚخ نهفئشا ان ظًبثخ ف انذساعبد انزششٚذٛخ. ثٔبنزبنٙ ٚ کً انز طٕٛخ ثأ فطش ػٛش انغشاة اکزغت أ ًْٛخ
رغز ٚٔخ ػٔلاجٛخ أػذخ ٚٔ کً اػبفز ثظ سٕ يز ػُٕخ نغزاء ان شًػ ان ظًبث ثبنزغ ىً
أنزهٛف انکجذ .ٖ
انزغ ىً انکجذ , انفئشا .ٌ , Agaricus bisporus الکلمات المفتاحية : فطش ػٛش انغشاة
142 Egypt. J. of Appl. Sci., 35 (11) 2020