Ankylosing Spondylitis (AS) is a chronic rheumatic disease that principally affects the intervertebral and sacroiliac joints. Two major features of AS are inflammation and bone reformation. Th17 cells as a new subpopulation of CD4+ T cells, are characterized by the production of pro-inflammatory cytokines. Th17 cells have been implicated in autoimmune diseases, pathogenesis and diagnosis of several inflammatory diseases, such as AS. Regulatory T cells (Treg) with suppressive effects on inflammation and autoimmunity have been reported to implicate in pathology of AS. The Treg /Th17 functional balance is essential for the prevention of autoimmune and inflammatory diseases by preventing deleterious impairment to the host and mounting effective immune responses. A group of circulating miRNA in plasma is found to be the change they can be involved in inflammation or inhibit it. miRNAs have been shown to play a pivotal role in the pathogenesis of various diseases including autoimmune or auto-inflammatory diseases.The function and molecular pathways of several key deregulated miRNAs, are elucidated in AS patients. Curcumin is an active component of turmeric which is a perennial plant. Curcumin is able to exert anti-atherogenic, anti-cancer and anti-inflammatory effects. The curcumin induces down-regulation of various inflammatory cytokines including TNF-α and IL-1. The solubility of curcumin in nanomicelles spherical water increases to more than 100 thousand times, which significantly enhances the absorption of curcumin. The aim of the present study was to understand the nano-curcumin effects on 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 AS and their correlation with the disease progression.
A16-weeks randomized placebo-controlled study was conducted on a total of 24 patients with age range of 22 to 50, who were clinically diagnosed with ankylosing spondylitis on the basis of clinical manifestations. The AS patients were divided into 2 subgroup with a block randomization, 12 out of 24 received a daily dose of 80 mg oral nano-curcumin and 12 patients received placebo as control group in a period of 4 months. Peripheral blood samples (8 ml) were obtained from the patients in both control and treatment groups before and after nano-curcumin treatment for 4 months. PBMCs were isolated from samples using Ficoll separation technique. Subsequently, cells were cultured in the presence of PMA. Treg cells associated immunological parameters such as mRNA expression levels of mir-146a, mir-27 and mir-17, TGF-β, IL-10, IL-6 and FoxP3 and alsoTh17 related immunological parameters such as mRNA expression of mir-141, mir-155 and mir-200, IL-17, IL-23 and RORγt were measured by real-time PCR, also Treg and Th17 frequency and their related cytokines secretion levels were evaluated respectively by flowcytometry and ELISA technique in both groups, pre and post-treatment.
Study Type
INTERVENTIONAL
Allocation
RANDOMIZED
Purpose
TREATMENT
Masking
DOUBLE
Enrollment
24
Nanocurcumin capsules (the formulation of curcumin nanoparticles, Exirnanosina). Subjects randomized to Nanocurcumin Arm will receive 80 mg/day for 4 months
Subjects randomized to Placebo Arm will receive placebo in the form of capsules for 4 months
Connective Tissue Diseases Research Center
Tabriz, Iran
Assessments of Ankylosing Spondylitis Signs and Symptoms (BASDI)
Number of Subjects With a Reduction in Signs and Symptoms
Time frame: 4 months after treatment
mir-141, mir-155 and mir-200 expression
qPCR method (mir-141, mir-155 and mir-200 induces differentiation of Th17 cells and increase inflammation)
Time frame: 4 months after treatment
Serum IL-17 levels
Elisa method (Th17 cells produce inflammatory cytokine, IL17, and increase inflammation).
Time frame: 4 months after treatment
RORγt expression
qPCR method (RoRγt, a transcription factor, induce Th17 cell differentiation and increase inflammation).
Time frame: 4 months after treatment
IL-17 expression
qPCR method (Th17 cells produce inflammatory cytokine, IL17, and increase inflammation).
Time frame: 4 months after treatment
Th17 cells frequency
Flowcytometry (Th17 cells produce inflammatory cytokine, IL17, and increase inflammation).
Time frame: 4 months after treatment
mir-27, mir-17 and mir-146a expression
PCR method (mir-27, mir-17 and mir-146a induces differentiation of Treg cells)
Time frame: 4 months after treatment
Serum TGF-β, IL-10, IL-6 levels
Elisa method (Treg cells produce anti-inflammatory cytokine, and decrease inflammation).
Time frame: 4 months after treatment
FoxP3 expression
qPCR method (FoxP3, a transcription factor, induce Treg cell differentiation and decrease inflammation).
Time frame: 4 months after treatment
TGF-β, IL-10, IL-6 expression
qPCR method (Treg cells produce anti-inflammatory cytokine, and decrease inflammation).
Time frame: 4 months after treatment
Treg cells frequency
Flowcytometry (Treg cells produce anti-inflammatory cytokine, and decrease inflammation).
Time frame: 4 months after treatment
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