Introduction: The microRNA-326 (miR-326) gene, by targeting ETS Proto-Oncogene 1 (ETS1), regulates the differentiation and interleukin-17A production of T helper 17 (Th17) cells. Celiac disease (CD) is an intestinal autoimmune disorder, in which the cascade of Th17 cells plays an important role in its pathogenicity. The aim of this study was to evaluate the expression changes of miR-326 and its two target genes ETS1 and IL-17A in celiac disease patients under a gluten-free diet (GFD). We expected the expression of miR-326 and IL-17A gene to decrease, and the expression of the ETS1 gene to increase, following the adherence to GFD. Methods: Peripheral blood samples of 40 CD patients under GFD (for more than 1 year) and 40 healthy individuals were collected. RNA was extracted, cDNA was synthesized and the miR-326, ETS1 and IL-17A gene expressions were evaluated by the quantitative polymerase real-time qPCR method. P-value ˂ 0.05 was considered statistically significant. Results: Although miR-326 mRNA expression was significantly lower in CD patients (P = 0.001), no significant difference was observed in ETS1 mRNA level between the two groups (P = 0.54), but IL-17A was significantly overexpressed in CD patients (P=0.002). No significant correlation was observed between the expression of the studied genes and the patients’ symptoms and Marsh classification. Conclusion:Adherence to the GFD for one to two years did not have the expected effect on the expression of genes in this panel. The most important finding that contradicted our hypothesis was the observation of high IL-17A levels in CD patients despite dieting, which may be related to the protective effect of this cytokine on intestinal tight junctions, that needs to be confirmed in further studies.
Background. One of the effective genes in the pathogenesis of the celiac disease is the Ets1 gene, which encodes the transcription factor Ets1 and is highly conserved during evolution. The Ets1 gene inhibits the differentiation of T helper 17 (Th17) cells and the production of interleukin-17A (IL-17A) by these cells and decreased expression of the Ets1 gene can lead to autoimmune disorders. The aim of this study is to evaluate the changes in Ets1 gene expression in the blood samples of patients with celiac disease compared with the control group. Methods. Blood samples were collected from twenty patients with celiac disease under a gluten-free diet and also from twenty healthy people. After RNA extraction and cDNA synthesis, a specific primer pair of the Ets1 gene was designed and its expression changes were examined by real-time PCR. Results. The expression of the Ets1 gene in patients with celiac disease on a gluten-free diet did not show a significant difference compared with healthy individuals (p-value= 0.54).. Conclusion. Failure to observe a significant difference between the patient and the control group can be due to the effect of the duration of the gluten-free diet and also the inadvertent entry of gluten from hidden sources into the diet of patients under treatment. Practical Implications. According to the results observed in this study, it is possible that if the gluten-free diet is followed more strictly and over a longer period of time by patients with celiac disease, the expression of the Ets1 gene will proceed as we expected. This issue needs to be evaluated in future studies with a larger community.
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