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Molecular genetic studies on Tilapia fish

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Mohamed El-Adl

on 30 October 2013

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Transcript of Molecular genetic studies on Tilapia fish

A Thesis presented
By

Influences of dietary restrictions and age on liver enzymes activities and lipid peroxidation in rats
Aim of the work
Conclusion
This study aimed to compare the effect of dietary restriction on young and adult rats fed both control and caloric restricted diet for two months in order to investigate the effect of caloric restriction on animal health
Material and Methods
Mohamed Adel Lotfy EL-Kholy
Under supervision of
Prof. Dr
El-Said El-Sherbini El-Said
Professor and Head of the
Department of Biochemistry
and Chemistry of Nutrition,
Faculty of Veterinary Medicine,
Mansoura University
Prof. Dr.
Gehad Ramadan El-Sayed
Professor of Biochemistry and
chemistry of Nutrition,
Faculty of Veterinary Medicine,
Mansoura University
Results
Introduction
Dietary restriction:
A dietary regimen in which caloric intake is decreased by 30-40% and supplied with sufficient amounts of nutrients and vitamins
Thesis entitled
Examining committee
Prof. Dr.
Yousef Mohamed Aly Mostafa
Professor of Biochemistry,
Faculty of Veterinary Medicine,
Zagazig University
Prof. Dr
Yakout Abdel-Fattah El-Senosy,
Professor of Biochemistry,
Faculty of Veterinary Medicine,
Banha University
f
Caloric restriction is considered as a great regimen in improving the overall health through the improvement of
cardiovascular system
and reducing the
oxidative stress markers
such as hydrogen peroxide, protein carbonyls, inflammatory cytokines as TNF-a, IL-6, C-reactive protein, and NF-β and increase antioxidant capacity which accompanied with the increase in antioxidant defenses of the body
The aging process
is an acceleration of the organ functional decline which is explained by mitochondrial free radical theory of aging stated that
"The increase in oxygen radical production with age through a leakage in mitochondrial system, causing oxidative stress,
lipid peroxidation
, and aging in the cells. Caloric restriction reduces the adverse effects associated with aging" which is illustrated as follows:
Lipid peroxidation
is an important and probably primary event in the progress of several diseases as well as degenerative processes associated with aging.The products of lipid peroxidation initiate further free radical chain reactions and can cause oxidative damage to lipids,proteins, and DNA.
Forty rats were subdivided into 8 equal group as follows:
AC2: five adult rats fed on control diet and scarificed after the
second month.
AC1: five adult rats fed on control diet and scarificed after the
first month
YR2: five young rats fed on 40% caloric restricted diet and scarificed after the
second month
.
YR1: five young rats fed on 40% caloric restricted diet and scarificed after the
first month.
YC2: Five young rats fed on control diet and sacrificed after the
second month
YC1: Five young rats fed on control diet and scarificed after the
first month
AR1: five adult rats fed on 40% caloric restricted diet and scarificed after the
first month.
AR2: five adult rats fed on 40% caloric restricted diet and scarificed after the
second month.
Experimental design was carried out as follows:
After the end of the experiment, rats were anesthetized using thiopental sodium via intraperitoneal route and blood samples were collected from the ventricles of heart
Blood samples were divided into 2 aliquots as follows:
The first part was used as a heparnized blood (20 IU/ ml), which is further subdivided into 2 parts
The second part of blood samples were left to clot for obtaining serum which was used for:
Determination of MDA concentration
Determination of lipid profile
Upper plasma layer for determination of plasma catalase activity
Lower erythrocytic layer was lysed using cold distilled water and used for:
Determination of blood SOD activity
Determination of blood GPx activity
Rats were dissected and liver samples was obtained, washed by normal saline and stored into two parts as follows:
The first part was stored at -20ºC for determination of:
Tissue SOD activity
Tissue CAT activity
Tissue GPx activity
Tissue MDA levels
The second part was stored at 20% formalin for immunolocalization of GPx in liver of rats and histopathological examination of liver to reveal the effect of caloric restriction
Statistical analysis was performed according to
steel and Torrie
utilising:
The mean and its standard error.
Least significant deference (L.S.D. at 0.05 level).
Analysis of variance using one way classification.
The calculation of statistics was done by SPSS V.17 software
Effect of dietary restriction and age on the activity of SOD in erythrocyte lysate of rats (U/ml).
Effect of dietary restriction and age on liver SOD activity of rats (U/gm)
Effect of dietary restriction and age on the activity of CAT in plasma (U/L) of rats.
Effect of dietary restriction and age on the activity of CAT in liver (U/gm) of rats.
Effect of dietary restriction and age on the activity of GPx in erythrocyte lysate of rats (U/ml).
Effect of dietary restriction and age on the activity of GPx in liver (U/gm) of rats.
Effect of dietary restriction and age on the level of MDA activity in serum (umol/L) of rats.
Effect of dietary restriction and age on levels of serum total cholesterol (mg/dl) in rats.
Effect of dietary restriction and age on the level of serum HDL-Cholesterol (mg/dl) in rats.
Effect of dietary restriction and age on levels of LDL-Cholesterol serum (mg/dl) in rats.
Effect of dietary restriction and age on the levels of serum Triacylglycerol (TAG) (mg/dl) in rats.
Effect of dietary restriction and age on serum total lipid levels (mg/dl) in rats.
The result of this study stated that the caloric restricted diet would result in a significant increase of hepatic and whole blood antioxidant enzymes, especially when performed in young age. Moreover, caloric restriction showed a significant depression of lipid peroxidation in rats.
Effect of dietary restriction and age on the level of MDA concentration in liver tissue (nmol/gm) of rats:
Histopathological studies
Immunohistochemistry for liver GPx localization
Full transcript