Evaluation of HMGB1 and HMGB2 effect on expression of TLR2,TLR4 and TLR9 from PBMC of MS patient

Abstract

MS is an inflammatory disease in which the myelin sheaths of nerve cells in the brain and spinal cord are damaged. This injury can impair the ability of the parts of the nervous system that are responsible for communication and cause many physical signs and symptoms. In addition to blood cells and tissue epithelial and endothelial cells, TLRs are also present in the brain.. HMG proteins are thought to play an important role in various human disorders. The reason for choosing HMG is due to this inflammatory modulatory property in this study and the reason for choosing TLR2, TLR4, TLR9 is due to the great importance of these 3 receptors in transmitting messages at the cell surface and membrane surface. Materials & Methods: 30 MS patients referred to Imam Reza MS Clinic with mild to moderate stages were selected and blood samples were taken from patients and MACS monocytes were isolated from PBMC. Monocytes were evaluated for purity percentage by flow cytometry. Was treated with CD14 antibody. Monocyte cells were then cultured and stimulated with HMGB1 and HMGB2 proteins at time, zero, six, twelve, and twenty-four and forty-eight hours. After stimulation, cells were collected and the expression levels of TLR2, TLR4, and TLR9 were evaluated by Real Time PCR. Results: The expression level of TLR2 and TLR4 receptors increased significantly with time course of stimulation time, which reached its highest value in 24 hours after stimulation with HMGB2. Compared to TLR2 and TLR4, the increase was the same. The performance of HMGB1 was evaluated as a time course that the expression of TLR2, TLR4, TLR9 showed a significant increase over time that this increase increased significantly in 24 hours after stimulation with HMGB1.