الفهرس 
Oxidative Stress in Liver Diseases
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Abstract
Liver disease is a chronic disease in many countries of the world,
due to lack of effective protective / therapeutic option for the disease. It
has been reported that a high percentage of Egyptian society has liver
disease, which is mainly due to chronic hepatitis C virus (HCV), in
addition to other risk factors including obesity, diabetes, excessive
medication and environmental pollution.
Oxidative stress is a common mechanism involved in the initiation
and progression of liver damage in different types of liver disease. It has
been found that the use of natural antioxidants is useful in the plan of
logical therapeutic in the prevention and therapeutic of liver diseases
including oxidative stress. There is a trend in the search for natural
antioxidants of natural sources that have a high efficiency in the
protection and therapeutic of liver diseases.
Moringa Oleifera Lam (M.O) a plant native to Indian, and then
spread to many parts of the world, and the different parts of this plant
have been used in ancient Indian civilization as an important food source
and in the therapeutic of some diseases, and recently found that the
different parts of the plant contain high levels of vitamins and minerals
that are useful in food, in addition to containing antioxidants such as
Phenols and flavonoids, which can be used in the protection or
therapeutic of diseases that involve oxidative stress.
Acetaminphen (Paracetamol) APAP is most widely used in the
world as an analgesic and antipyretic drug that is safe at therapeutic
dosages. Paracetamol is available as oral, rectal and injectable
formulation. Paracetamol is prescribed as pain reliever and fever reducer.
It is commonly used to relieve headache and other minor muscle aches,
pain and major ingredient in numerous cold and flu remedies, and broadly
use in the management of more severe pain such as post-surgical pain and
providing palliative care in advanced cancer patient. The use of Paracetamol at recommended doses is safe. However, its use at high doses
causes significant damage to the liver. Therefore, the models of liver
damage induced by Paracetamol in rats and mice are widely used to study
the effects of antioxidants in liver protection.
This study aimed to:
(1) Evaluation of antioxidant activity of different parts (leaves, flowers,
stem bark, roots, seeds) of Moringa Olifeira plant extracts in-vitro
and to identify the parts containing high antioxidant activity
(2) Evaluation of ethanolic extracts of leaves and pods of Moringa
Oleifera as protective against paracetamol-induced liver damage
(acetaminophen -APAP) in rats. In this experiment, the ethanolic
extract, for leaves or stem bark of the Moringa (300 mg in H2O +
DMSO / kg of rats weight or 600 mg in H2O + DMSO / kg of rats
weight) were given to the rats one hour before to administration of a
single dose (oral) of acetaminophen (APAP) dissolved in DMSO
(4g/kg bw of the rat). Protective activity of Moringa extracts of
leaves and pods was followed weekly for 21 days to monitor liver
functions enzymes such as alanine aminotransferase (ALT) and
oxidative stress markers such as Malondialdhyde (MDA), Catalase
(CAT) and the histological changes in the liver were compared
between groups of rats treated with Moringa extract with a group of
rats treated with acetaminophen alone. In this experiment, kidney
function markers (urea and creatinine) were also measured. As
oxidative stress in the liver is accompanied by oxidative stress in the
kidney.
For this purpose male albino 42 rats were divided into seven groups
(each group has 6 rats) as the below:group I: Normal control (receiving water (W) daily).
group II: Rats received (DMSO daily) control negative.
group III: Rats received a singl dose of 4g APAP/kg b.w once (positive
control).
group IV: Rats received 4g APAP/kg b.w once one hour after receiving
the M.O extract (1mL) 300 mg of M.O leaves extract/kg b.w,
daily, for 21 days
group V: Rats received 4g APAP/kg b.w once one hour after receiving
the M.O extract (1mL) 600 mg of M.O leaves extract/kg b.w
daily, for 21 days
group VI: Rats received 4 g APAP/kg b.w once one hour after receiving
the M.O extract (1mL) 300 mg of M.O stem bark extract/kg
b.w daily, for 21 days
group VII: Rats received 4 g APAP/kg b.w once one hour after
receiving the M.O extract (1mL) 600 mg of M.O stem bark
extract/kg b.w daily, for 21 days
The most important results obtained include the following elements:
1. Evaluation of the antioxidant activity (In -vitro) using the DPPH for
different parts of Moringa using three solvents: ethyl alcohol, water
and hexane. The results shown that the ethanolic extracts gives the
highest efficiency of the antioxidant activity of all parts of the plant.
In addition, it was found that the extract of flowers gave the highest
efficiency of antioxidant activity (94.91%) followed by leaves extract
(93.10%), stem bark (90.78%), roots (84.80%) and finally seeds
(62.15%). This is compared to Vit C (ascorbic acid) which used as
reference compound given (94.00%)
2. High-efficiency liquid chromatography – Mass spectroscoby (HPLCMS)
analysis for ethanolic extract of leaves detect five phenolic and
flavonoid compounds, including chlorogenic acid, Apigenin-8-C— glucoside, Quercetin-3-O-β-D-glucoside, Quercetin-3-O-acetyl
glucoside and kaempferol-3-O-glucoside. These compounds are
known to have anti-oxidant activity. Although there were other
compounds in the ethanolic extract of leaves, it was not possible to
identify them because there were no reference compounds for
comparison.
3. In the biological experiment in-vivo, the ethanolic extract of both
leaves and pods in doses (300 mg / kg of rats weight and 600 mg / kg
of rats weight) was given orally one hour before to the preoperative
dose of paracetamol (APAP) (4g / kg) of the rats. The preventive
activity of these extracts was studied, by monitoring the level of liver
function markers, oxidative stress markers and histopathology in the
liver, weekly for 21 days. The most important results obtained are:
(a) In the group of rats treated with acetaminophen (APAP) alone
(positive control or hepatotoxin control) there was a significant
increase ( p ≤ 0.05 ) in liver function enzymes ALT, AST, GGT,
ALP in serum at the first week and continued to increase until
the end of the experiment (at the third week) compared to the
normal control group. The change in these enzymes indicates
damage to the liver tissue due to oxidative stress of paracetamol
(APAP).
(b) In the group of rats treated with ethanolic extract of leaves and
stem bark in doses 300 and 600 mg / kg of rats weight (Groups 4
through 7) the results showed a significant decrease (p ≤ 0.05) in
the levels of these enzymes during the three weeks compared to
the group of positive control (paracetamol treatment alone). This
indicating the protective effects of both ethanolic extract of
leaves and stem bark, where the levels of these enzymes returned
to normal level at the end of the experiment. In addition, this
reduction depends on the amount of the dose of the extract.(c) The results also showed a significant increase in serum total
bilirubin and a significant decrease in serum total protein during
the three weeks in the group treated with paracetamol (APAP)
alone. These results also indicated liver tissue damage due to
paracetamol effects. On the other hand, the treatment with the
ethanolic extracts of both leaves and stem bark returned these
parameters to their normal level, which indicates their protective
effects on the liver.
4 - The study of the protective effect of the ethanolic extracts of leaves
and stem bark of Moringa on the oxidation markers that including
lipid oxidation (LPO) using the test of Malondialdhyde (MDA) and
the enzymes of the antioxidant superoxide dismutase (SOD) and
catalase (CAT).The results showed the following:
(a) In the paracetamol(APAP) group alone, the MDA level in the
blood increased after a week of treatment and continued to rise
steadly to the end of experiment. In comparison, the treatment
of ethanolic extracts of leaves and stem bark of Moringa
showed a significant decrease (p ≤ 0.05) in MDA during the
experiment period and the decrease reached a level slightly
lower than the normal control at the end of experiment. In
addition, the treatment with solvent DMSO led to a small
decrease compared to normal control.
(b) Treatment with paracetamol(APAP) significantly reduced SOD
and CAT at the end of the experiment, indicating oxidative
stress. On the other hand, the protective effects of both the
ethanolic extract of leaves and stem bark against the oxidative
stress of paracetamol (APAP) showed a high level of these
enzymes to a level higher than both the positive control and the
normal control.
5. Histological changes. The histological examination of the liver for different treatments showed
the following:
The normal control group showed no histopathological changes in
the liver, tissues at the end of the experiment.
The control group for DMSO also showed no histopathological
changes in the liver, tissues at the end of the experiment.
The positive control group (APAP treatment alone) showed a focal
death in the liver with inflammation of the cells, in addition to
hyperplasia in the superficial surface of the vertebrae, at the end of
the experiment.
The group treated with ethanolic extract of leaves and stem bark
showed slight improvement in these histological changes, at the end
of the experiment.
from these results we can concluded the following:
1- The ethanolic extracts of Moringa oleifera Lam leaves and
stem bark showed antioxidant activities (both in- vitro and invivo)
due to the presence of active phytochemical such as
phenols and flavonoids and other active compounds .
2- The ethanolic extracts of Moringa Oleifera Lam leaves and
stem bark exhibits free radical scavenging and
hepatoprotective and kidney protective against oxidative stress
injury.