Evaluation of High Mobility Groups on expression of TLR2 , TLR4 , and TLR9 in patients with Hydatid cyst

Abstract

Hydatidosis is a common disease between humans and animals. Due to the high prevalence of hydatid disease in Iran, especially in rural areas and the heavy health and economic burden of this disease, prevention, diagnosis and treatment of this disease is very important. However, there is no effective treatment for hydatidosis. In this study, the effect of HMG group proteins on the expression level of TLR2, TLR4 and TLR9 in patients with hydatid cyst was evaluated. Methods: Monocyte mononuclear cells were cultured in DMEM culture medium and extracted by MACS method. The percentage of cell isolation was evaluated by flow cytometry using CD14 antibody. It was stimulated with HMGA1, HMGA2, HMGB1, and HMGB2 separately as a time course with 0, 2, 6, 12, 24, and 48 hours. After RNA extraction and cDNA synthesis, the expression level of TLR2, TLR4, and TLR9 was evaluated by Real-Time PCR method. The inflammation was measured by the inflammatory cytokines IL-1 and IL-6 using the ELISA method. Statistical analysis was done using Graphpad prism 7 software. One-way ANOVA test was used to compare gene expression levels. P<0.05 was considered significant. Results: In this study, the expression of TLR2, TLR4, and TLR9 in the group of patients after exposure to HMG group proteins showed a statistically significant increase over time, and this increase was significant in 24 hours after stimulation. The amount of both IL-1 and IL-6 inflammatory cytokines increases to a statistically significant ratio in the patient group.