Determination of Methylation and expression of TLR2 Genes in Ptients with Behc¸et’s Disease and TLR4

Abstract

Considering the proven role of TLRs including TLR2 and TLR4 inrheumatic and Behcet’s disease and lack of study on the level of methylation andexpression of the genes involved in this pathway, we determined that the level ofmethylation and expression of TLR2 and TLR4 genes as one of the types ofreceptors involved in the pathogenesis of Behcet's disease, in order to provide thepossibility of identifying and identifying the possible mechanisms for the possibleassociation of these genes with Behcet’s disease.Methodology: In a case-control study, 47 Iranian Azeri BD patients and 61 ageandsex matched healthy controls recruited to the study. Peripheral bloodmononuclear cells (PBMCs) were isolated from EDTA blood tubes by Ficolldensity-gradient centrifugation. Genomic DNA samples of BD and healthycontrols were extracted using the rapid genomic DNA extraction method from theperipheral blood collected in tubes containing EDTA. Total RNA was extractedfrom the PBMCs according to the TRIzol protocol. MiR-146a, miR-155, TNF-αand CTLA-4 expression were studied using real-time PCR.Results: It was observed that the mean of TLR4 expression in the patient groupwas higher than the healthy group, which was statistically significant. While thelevel of methylation of TLR4 in the patient group was lower than that of thehealthy group, this difference was also significant. Also, the mean of TLR2expression in the control group was lower than that of the patients. This differencewas not statistically significant between the healthy subjects and those withBehcet's disease. Moreover, the methylation no significant difference betweenpatients and healthy TLR2 also was found.