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EFFECT OF EXERCISE AND/OR ANTIOXIDANT ON KLOTHO GENE MESSEN

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hala anwar

on 6 May 2014

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Transcript of EFFECT OF EXERCISE AND/OR ANTIOXIDANT ON KLOTHO GENE MESSEN

Aim of the work
This work aimed at clarifying the effects of exercise and/or antioxidants on klotho gene messenger RNA expression during oxidative stress induced by obesity in renal tissue of adult male albino rat.
EFFECT OF EXERCISE AND/OR ANTIOXIDANT ON KLOTHO GENE MESSENGER RNA EXPRESSION DURING OXIDATIVE STRESS IN RENAL TISSUE OF ALBINO RATS
By, Hala Magdy Anwar
Acknowledgement
Effect of consumption of high fat diet for 16 weeks (group IIa) & comparing it with control group(group I):
There was significant increase in body weight.

There was significant increase in obesity-induced oxidative stress markers.
Significant decreased in Klotho gene expression in the kidney.
On increasing the duration of consumption of high fat diet for 24 weeks (groupIIb) (+ve control)& comparing it with group that consumed high fat diet for 16 weeks(groupIIa):
There were non significant changes in different parameters except for significant increases in serum glucose and TNF alpha expression levels in the kidney in the group recieving high fat diet for 24 weeks (group IIb +Ve control
Effect of exercise training for 8 weeks (groupIII) &comparing it with rats receiving high fat diet for 24 weeks(groupIIb):
Significant decreases in body weight.

Significant increase in obesity-induced oxidative stress markers.

Significant increase in Klotho gene expression level in the kidney.
Effect of antioxidant treatment for 8 weeks (groupIV) &comparing it with rats receiving high fat diet for 24 weeks(groupIIb):
Significant decreases in serum glucose level & HOMA-IR
Non significant change in body weight,serum insulin,serum creatinine and TNF alpha gene expression levels in the kidney.
Significant increase in Klotho gene expression level in the kidney.

Effect of combined exercise training &antioxidant treatment for 8 weeks (groupV) & comparing it with rats receiving high fat diet for 24 weeks(groupIIb):
Significant decreases in obesity-induced oxidative stress markers.
Significant increase in Klotho gene expression level in the kidney.
serum insulin
HOMA
TNFa log 10
serum creatinine
serum glucose
Weight
Significant decreases in plasma insulin level among exercise group and combined exercise& antioxidant group when compared with high fat diet group with the more reduction was among combined exercise& antioxidant group.
Significant decreases in HOMA-IR among exercise group, antioxidant group and combined exercise& antioxidant group when compared with high fat diet group with the maximum reduction was among combined exercise& antioxidant group.
Non significant change regarding serum creatinine level among exercise group ,antioxidant group and combined exercise& antioxidant group when compared with high fat diet group.

Significant decreases in the level of TNFα mRNA expression level in the kidney among exercise group and combined exercise& antioxidant group when compared with high fat diet group with the more reduction was among exercise group.

Significant decreases in body weight in exercise group & combined exercise and antioxidant group when compared with high fat diet group with more weight loss was among combined exercise training & antioxidant group.
Significant decreases in serum glucose level among exercise group, antioxidant group & combined exercise& antioxidant group when compared with high fat diet group with the maximum reduction was among combined exercise& antioxidant group.
Significant increases in the level of Klotho mRNA expression level in the kidney among exercise group, antioxidant group and combined exercise& antioxidant group when compared with high fat diet group with the highest increment was among group combined exercise& antioxidant group.

Group (I): (-ve control group) received balanced diet and olive oil daily orally for 16 weeks
Group (II) was subdivided into:

Group III (exercise group): received high fat diet and olive oil daily orally for 24 weeks and were submitted for exercise training in the last 8 weeks.
Group (IV) (antioxidant group): received high fat diet for 24 weeks and received vitamin E in the last 8 weeks.
Group (V) (exercise and antioxidant group): received high fat diet for 24 weeks and were submitted for exercise training and received vitamin E daily orally in the last 8 weeks.
IIa: received high fat diet and olive oil daily orally for 16 weeks.
IIb (+ve control): received high fat diet and olive oil daily orally for 24 weeks
The study period lasted for 24 weeks which includes:
16 weeks for induction of obesity mediated an oxidative stress.
8 weeks for exercise training and treatment with antioxidant.

At the end of the study we measure body weight of each rat.
Klotho gene expression was detected in the kidney using RT-PCR.
The parameters used to evaluate the HFD induced oxidative stress were:
Serum glucose.
Serum insulin.
HOMA-IR.
Serum creatinine.
TNF expression in renal tissue using RT-PCR.
Post-run Amplification Plots of control, high fat diet ,exercise, antioxidant and combined exercise &antioxidant Groups for TNFα mRNA as Selected Target Detector and GAPDH mRNA as Housekeeping Detector (Endogenous Control).
Post-run Amplification Plots of control, high fat diet ,exercise, antioxidant and combined exercise &antioxidant Groups for TNFα mRNA as Selected Target Detector and GAPDH mRNA as Housekeeping Detector (Endogenous Control).
There were significant -ve correlations between KLOTHO mRNA and TNFα mRNA in all groups.

Material And Method
Results
Results
Results
Conclusion
We conclude that the exercise training and /or antioxidant treatment improves HFD induced oxidative stress in renal tissue and up-regulates mRNA expression of klotho gene in the kidney and this suggests that the effect of the exercise training and /or antioxidant treatment on oxidative stress may be mediated even partially through increased klotho mRNA gene expression. And we assume that this role of klotho may be mediated through downregulation of TNF gene expression
This study was conducted on 42 adult male albino rats, weighing between 170 and 200 g. The animals were randomly divided into 5 groups each consisted of 7 rats as follow:
Oxidative stress occurs when the balance between antioxidants and reactive oxygen species (ROS) are disrupted because of either depletion of antioxidants or accumulation of ROS.
Oxidative stress balance
Obesity
Vitamin E
Excercise
Kidney
The most well-researched non-enzymatic antioxidants.
Acts as a free radical sweeper, preventing the propagation of free radicals in tissues.
Being fat-soluble, it is incorporated into cell membranes, protecing them from oxidative damage
Protects lipids and prevents the oxidation of polyunsaturated fatty acids
Regular training is known to increase the resistance against ROS induced lipid peroxidation, and to decrease the accumulation of oxidative protein and DNA damage.
This may occur through enhancement of intrinsic antioxidant potential or adaptation to oxidative stress that occurs in trained individuals
Obesity has been described as a state of chronic oxidative stress. Furthermore, oxidative stress has been defined as the link between obesity and its major associated disorders such as insulin resistance, hypertension, etc
The kidney is an organ highly vulnerable to damage caused by ROS, likely due to the abundance of long-chain polyunsaturated fatty acids in the composition of renal lipids
Klotho gene
Overexpressed
Downexpressed
kidney
=
Antioxidants
=
oxidants
Klotho gene is a transmembrane protein that functions as an aging -suppressor gene that extends life span when over expressed and accelerates aging-like phenotypes when disrupted in mice, also it protects cells and tissues from oxidative stress.
Klotho is predominantly expressed in the kidney, with its highest expression in cells of the distal convoluted tubule.
Studying the effect of longer duration of exercise training (>4 weeks) on the same parameters and whether this improvement would be reversed after stoppage of exercise.

Studying the effect of longer duration of vitamin E supplementation (>4 weeks) on same parameters and whether this improvement would be reversed after stoppage of the intake of vitamin E.
Studying of other benefits of Klotho gene such as antiaging activity.
Further studies on comparison between effects of exercise training between young aged and old aged rats.
Further studies on comparison between effects of vitamin E supplementation between young aged and old aged rats.
Recommendations
Recommendations
Recommendations
Recommendations
Recommendations
Tumor Necrosis factora (alpha TNF-a)
An inflammatory cytokines that has been used as a marker to correlate effect of oxidative stress locally in the kidney biopsies.
Oxidative stress increases TNF-a expression
Antioxidants decreases TNF-a expression
TNF-a was also used to correlate with Klotho gene expression
in the high fat diet consumption group (IIa)
In the exercise group (III)
In antioxidant group
In combined exercise and antioxidant group (group V)
Thank you
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