The Effect of Melatonin on Gene Expression and Activity of the Sirt1 and Its Target Genes Catalase and MnSOD in Multiple Sclerosis Patients and Healthy Subjects

Study Overview
=================
Brief Summary
-----------------
The purpose of this study is to investigate the gene expression and activity of SIRT1, and forkhead box O3 pathway by means of investigating target genes MnSOD and catalase in multiple sclerosis and healthy subjects. Also, we investigate the effect of melatonin on interaction between SIRT1 activity and target genes MnSOD and catalase in multiple sclerosis and healthy subject .Additionally, we evaluate effect the probable effect of melatonin treatment on gene expression and activity of above-mentioned genes in peripheral blood mononuclear cells of healthy subjects-treated with H2O2.

Conditions
-----------------
Multiple Sclerosis, Oxidative Stress

Intervention / Treatment
-----------------
* Drug: Melatonin
* Other: Hydrogen peroxide


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria: patients with relapsing-remitting multiple sclerosis. The diagnosis of MS was based on the McDonald criteria . All patients were newly diagnosed cases with an Expanded Disability Status Scale (EDSS) score ≤ 6 and had never received immunomodulatory and immunosuppressive drugs. - Exclusion Criteria: The exclusion criteria were as follows: 1) treated with antioxidant supplements, anti-inflammatory drugs, and vitamins in the previous 6 months; 2) had a history of acute or chronic infection, malignancy, diabetes (type I and II), and any clinically significant systemic disease; 3) was currently pregnant or lactating; 4) was currently smoking; 5) used contraceptive medication during the previous 6 months.

Ages Eligible for Study
-----------------
Minimum Age: 20 Years
Maximum Age: 40 Years

Sexes Eligible for Study
-----------------
Female

Accepts Healthy Volunteers
-----------------
Accepts Healthy Volunteers

Study Plan
=================
How is the study designed?
-----------------

Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Single Group Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Multiple slerosis<br>Melatonin,,one millimolar,one micromolar,one nanomolar, 12 hr treatment. | Drug: Melatonin<br>* Melatonin is used for healthy subjects and multiple sclerosis patients<br>* Other names: N-acetyl-5-methoxy tryptamine;|
| Experimental: Healthy subjects<br>Melatonin,one millimolar,one micromolar,one nanomolar, 12 hr treatment. hydrogen peroxide, 250 micromollar,2 hr treatment | Drug: Melatonin<br>* Melatonin is used for healthy subjects and multiple sclerosis patients<br>* Other names: N-acetyl-5-methoxy tryptamine;Other: Hydrogen peroxide<br>* Hydrogen peroxide is only used for healthy subjects<br>|

What is the study measuring?
-----------------
Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Relative gene expression by real-time PCR (polymerase chain reaction) |  | Change from baseline after 12-hour treatment with melatonin and 2 -hours treatment with H2O2 | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| MnSOD enzyme activity assay |  | Change from baseline after 12-hour treatment with melatonin and 2 -hours treatment with H2O2 | 
| Catalse enzyme activity assay |  | Change from baseline after 12-hour treatment with melatonin and 2 -hours treatment with H2O2 | 
| SIRT1 enzyme activity assay |  | Change from baseline after 12-hour treatment with melatonin and 2 -hours treatment with H2O2 | 

 Terms related to the study
=================
Keywords Provided by Centre Hospitalier Valida
-----------------
SIRT1, Multiple sclerosis, MnSOD, Catalase, Oxidative stress, Antioxidant enzyme