RESUMO
STATEMENT OF THE PROBLEM: Oral squamous cell carcinoma is the most common oral malignancy. Toll-like receptor (TLR) activation led to alterations in the levels of mRNA encoding the TLR accountable for recognizing the inducing agonist and cross-regulation of other TLR. PURPOSE: The purpose of this study is determination of mitogen-associated protein kinase (MAPK) activation in human immortalized oral epithelial cell (HIOEC) line via up regulating of TLR7. MATERIALS AND METHOD: expression of TLR7 was measured in HIOEC and normal cells by quantitative real-time polymerase chain reaction (qRT-PCR) and samples were calibrated by ß-actin. RESULTS: Western blot analysis discovered high expression of TLR7 and MAPK in HIOEC cell lines. TLR7 was over-expressed in HIOEC cell line. Imiquimod-induced expression of interleukin (IL)-6, IL-8, and vascular endothelial growth factor (VEGF) was inhibited by TLR7 siRNA in HIOEC cells as determined by reverse transcription polymerase chain reaction (RT-PCR). Mean fluorescence intensity of nuclear p38 expression was determined in HIOEC cell lines (p< 0.05). RT-PCR analysis of IL-6, IL-8, and VEGF mRNA expression in HIOEC cells stimulated with imiquimod (1 µg/ml) for indicated time points. CONCLUSION: TLR7 is functionally over-expressed in HIOEC cell line of oral squamous cell carcinoma and development of resistance to cisplatin in human oral squamous cell carcinoma might occur through the mechanism involving activation of TLR7 and its dependent signaling pathway.
RESUMO
STATEMENT OF THE PROBLEM: Quercetin is a pharmacological flavonoid that can inhibit high mobility group box1 (HMGB1) protein, a non-histone nuclear protein that is implicated in inflammation. Th17 cells are important cells in the pathogenesis of inflammation. Pulpitis is the inflammation of dental pulp, which usually is accompanied by pain. Quercetin may alleviate this inflammation. PURPOSE: The current study aimed to compare blocking of HMGB1 function and stimulation of HMGB1 function with quercetin and investigate the effects of the blockage on T helper 17 (Th17) cells and mitogen-activated protein kinase Toll-like receptor 4 (MAPK-TLR4) signaling pathway. MATERIALS AND METHOD: T cells isolated from the pulp involved with pulpitis and the normal pulp were cultured. The cells suspensions were plated in 6-wells culture plates and stimulated with 0.5 µg/ml of HMGB1 for 2, 4, 8, and 12 hours. For blocking TLR4, 10 µg/ml rabbit anti-human TLR4 antibody was added 1 hour before treatment with HMGB1. RESULTS: The level of these cytokines decreased; moreover, western blot data showed that quercetin could decrease MAPK signaling pathway by means of inhibition of HMGB1 on T cells. The results showed the reduction of TLR4 pathway and Th17 cell polarization. CONCLUSION: Our results indicated that the levels of IL-17, IL-33, and IL-6 in supernatants from patients' cultured T cells were increased after stimulation with HMGB-1 following employing quercetin. It also could inhibit MAPK signaling pathway, which subsequently could decrease Th17 production and IL-17. Quercetin could decrease pro-inflammatory cytokines and IL-17 production.
