The Immunomodulatory Effects of Oral Nanocurcumin in Multiple Sclerosis Patients

Study Overview
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Brief Summary
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Multiple sclerosis is the most common autoimmune disease of the central nervous system, most ranging in age from 40-20 years of age is associated with neurons inflammation and demyelination. Increasing aggressive activities of Th17 and Th1 cells that their function is to secrete proinflammatory cytokines and decreasing the number and activity of regulatory T cells, which normally leads to controlling inflammation, are seen in these patients.Many studies have carried out to assess the prevalence of Tregs and Th17 in autoimmune disorders such as MS. The Treg /Th17 functional balance is necessary for the impediment of autoimmune and inflammatory diseases by preventing harmful injury to the host and increasing effective immune responses. miRNAs have been shown to play a pivotal role in the pathogenesis of various diseases including autoimmune or auto-inflammatory diseases. Curcumin, the active principle constituent of turmeric, is proved to be capable of regulating cellular responses and the growth of different cell types in the immune system such as B cells, T cells, macrophages, dendritic cells and natural killer cells. Curcumin has a combination of activities such as anti-inflammatory, antioxidant, anti-proliferation, anti-invasive, and can used in the treatment of Alzheimer's, Parkinson's, Multiple sclerosis, Cardiovascular disease, Bacterial diseases and Arthritis. The solubility of curcumin in nanomicelles spherical water increases to more than 100 thousand times, which significantly enhances the absorption of curcumin. The present study aimed at investigating the effects of nanocurcumin on the frequency of Treg and Th17 cells, expression levels of their associated transcription factors and cytokines, secretion levels of their associated cytokines and also related miRNAs expression levels in peripheral blood of patients with MS. .

Detailed Description
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In this study, fresh blood samples were acquired from 50 MS patients introduced by corresponding physician and Neurologist. As the aim of the current study was the assessment of nanocurcumin in MS patients, the peripheral blood collection was collected in two steps. After the first blood collection via venipuncture, 25 patients received nanocurcumin capsules each day and the remaining 25 patients took placebo for 6 months. Accordingly, the second sampling from the mentioned population was collected after 6 months. 9 patients were excluded from the study: 3 did not provide informed consent, 5 lived quite far away from the place in which follow-up examinations were performed, and 1 patient developed cancer. In all cases, 15 ml of blood was utilized for peripheral blood mononuclear cells (PBMC) isolation. PBMC is used to measure the expression levels of miRNA-106b, miRNA-25 and miRNA-326. In this regard, specific kits extracted miRNA from cells is used. Then the version of miRNA using Quantitative Real time polymerase chain reaction(qPCR).Then the frequency of Treg and Th17 cells in peripheral blood of patients will be examined by Flowcytometry and compared to the control group. The expression of key transcription factor (Foxp3) and specific cytokines (TGF-β) related to the Treg cells and key transcription factor (RORγt) and specific cytokines, interleukin-17(IL-17), related to the Th17 are measured by Quantitative Real time PCR. Elisa methos is used to assess the amount of the cytokine TGF-β and IL-17 in cultured cells.

Official Title
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The Effects of Oral Nanocurcumin on Expression Levels of microRNAs and Treg Cells and Th17 Cells Development Factors in Multiple Sclerosis Patients

Conditions
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Multiple Sclerosis

Intervention / Treatment
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* Drug: Nanocurcumin
* Drug: Placebo


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Willingness to cooperate Aged 18 to 65 years The diagnosis of Multiple sclerosis by Neurologist Patients in Relapsing Remitting (RRMS) Patients with Expanded Disability Status Scale (EDSS) <5/5. Exclusion Criteria: Use of nutritional supplements and antioxidant and immunosuppressive drugs alpha-lipoic acid a month before the study. Pregnancy and lactation History of diabetes and other chronic diseases History of other autoimmune diseases Occurrence of relapses during the study period Acceptance rate of less than 70% of supplements Unwillingness to continue to cooperate

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 65 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Patients who received nanocurcumin<br>Patients who received nanocurcumin | Drug: Nanocurcumin<br>* Patients will take 80 mg nanocurcumin in the form of capsules daily during the 6 month study period<br>* Other names: Patients who received nanocurcumin;|
| Placebo Comparator: Patients who received placebo<br>Patients who received placebo | Drug: Placebo<br>* Patients will take placebo in the form of capsules daily during 6 months study period<br>* Other names: Patients who received placebo;|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| EDSS measurment | EDSS measurment by neurologist | 6 months after treatment | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Treg cells frequency | Flowcytometry (Treg cells produce anti-inflammatory cytokines) | 6 months after treatment | 
| Th17 cells frequency | Flowcytometry (Th17 cells produce inflammatory cytokine and increase inflammation) | 6 months after treatment | 
| IL-17 and RORγt expression | qPCR method | 6 months after treatment | 
| IL-17 secretion levels | ELISA method | 6 months after treatment | 
| microRNAs (miRNA-326) expression | Evaluate the diagnostic value of microRNAs in quantitative polymerase chain reaction (qPCR), in MS patients as compared with healthy control | 6 months after treatment | 
| TGF-β and FoxP3 expression | qPCR method | 6 months after treatment | 
| TGF-β secretion levels | ELISA method | 6 months after treatment | 
| microRNAs (miRNA-106b and miRNA-25) expression | Evaluate the diagnostic value of microRNAs in quantitative polymerase chain reaction | 6 months after treatment | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Multiple sclerosis, Nanocurcumin, MicroRNAs, Th17 cells, Treg cells