Determination of Methylation and expression of IL-10 and IL-6 Genes in ptients with Behe ,ets Disease

Abstract

Introduction: Interleukin-10 (IL10) and interleukin-6 (IL6), produced generally by monocyte, T helper type 2 (Th2), and regulatory T cells (Treg), plays a central role in controlling inflammatory responses and regulating the immune response of the IL-10 mRNA expression, such IL10 is a reducing agent and IL6 an inflammatory augmentation. Although in most autoimmune diseases, such as Behçet’s disease, it has been proven that the expression of these genes is related to the disease process, no studies have been conducted on the role of epigenetic in regulating the expression of these genes in Behcet's disease. Therefore, the aim of this study was to evaluate the methylation and expression of IL10 and IL6 genes in patients with Behcet's disease and compare them with healthy subjects. Methods: This case-control study was performed on 51 patients and 63 healthy controls. Initially, the PBMC cells were isolated by ficoll method. Then DNA and RNA were extracted from all specimens. Promoter methylation levels of IL10 and IL6 were evaluated by MeDIP-qPCR technique. Also, the extraction of RNA was transformed into cDNA by RT-PCR technique and ultimately, IL10 and IL6 gene expression were measured by Real-time PCR. In the last section, they were evaluated for the relationship between gene expression and methylation, as well as their relationship with clinical specification. Results: As we expected, the expression level of the IL-10 gene was seen to significantly decline in the patient group in comparison to the control while the expression level of IL-6 gene was seen to significantly increase in the patient group compared to the healthy group. Also, the rate of promoter methylation was significantly higher in the IL-10 mRNA low expression group (patient group) compared to its high expression group (healthy group) (P < 0.001) while the relative promoter methylation level was significantly lower in the IL-6 mRNA high expression group (patient group) compared with its low expression group (healthy group) (p<0.001). Conclusion: According to the results of the expression and methylation of IL10 and IL6 in this study may be methylation of the promoter one of the regulatory mechanisms involved in the regulation of gene expression.