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Role of Toll Like Receptor 9 Signalling Pathway in Odontoblasts

Author: HuZuo
Tutor: YuQing；HeWenXi
School: Fourth Military Medical University
Course: Oral medicine
Keywords: Toll-like receptor 9 Dentin sialophosphoprotein Odontoblast
CLC: R781.1
Type: Master's thesis
Year: 2010
Downloads: 16
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Abstract

Caries is a the oral common disease. Cariogenic bacteria and / or toxic products when the carious dentin after arrive by dentinal tubules into the pulp cavity, resulting in pulpitis. Immune response, and the third of dentin pulp play defense and ability to repair. Odontoblast long cell processes into the dentinal tubules pulp dentin complex first point of contact for the bacterial microbial cells, then odontoblast and oral mucosal epithelial cells with innate immune function, no studies reported over the years. TLR9 innate immune receptors containing unmethylated CpG motifs in bacterial DNA (CpG DNA) specifically recognized in many tissue cells can be rapid activation of the innate immune response and the to start subsequent adaptive immune response, in the organization of cellular immune play an important role in the defense response, anti-infective, anti-allergic and anti-tumor therapy has made great progress. The Task Force found that the in vitro odontoblast-like cells expressing TLR9, prompted TLR9-mediated odontoblast natural immune function. But for odontoblast the body is to express TLR9 the TLR9 whether specific identification of bacterial DNA, TLR9 activation of intracellular signaling pathways downstream gene expression has not been reported. The face of bacterial invasion, in addition to the immune response, the formation of the third phase of dentin is the main form of pulp to play defense and the ability to repair another. Many studies have confirmed the the DSPP gene product in dentin formation and mineralization, and the third of dentin formation process plays a key role. DSPP gene expression in the process of bacterial infections result in odontoblasts or odontoblast-like cells to form a third of the dentin accompanied TLR9 signaling pathways activated by bacterial DNA into odontoblasts whether regulation of the DSPP gene expression at home and abroad has not been reported. In summary this experiment experimental methods of molecular biology the TLR9, DSPP gene research seeks to clarify the TLR9-mediated the odontoblasts natural immune response and regulation of gene expression of DSPP intracellular signaling pathways. Molecular mechanisms and DSPP investigate the caries progression bacterial DNA infection, into odontoblasts immune response, DSPP gene expression of dentin between clarify the cause of endodontics, pulp damage repair The law of regulation of gene expression is important for the immune the anticaries research and living pulp conservation therapy to provide new ideas and approaches. 1 TLR9, DSPP in odontoblasts and pulp tissue expression was the first to collect the pulp tissue in mice using tissue blocks RNA was extracted method to extract total RNA, reverse transcription, PCR amplification observed in the pulp tissue TLR9 mRNA expression; followed by extraction of total RNA from mouse OLC odontoblast cell line, reverse transcription PCR amplification observed TLR9 mRNA, DSPP mRNA expression. 1% agarose gel electrophoresis was TLR9 mRNA in the pulp tissue in mice and OLC cell lines were significant expression, DSPP mRNA in OLC cell lines revealed significant expression. TLR9 mRNA molecular weight 198bp, DSPP mRNA molecular weight 151bp. Of 2 CpG DNA OLC intracellular TLR9 DSPP gene expression and regulation of 6-well plates cultured the OLC cells stimulated with CpG ODN-A and CpG ODN-B cells, set the gradient 0,3,6,9,12,24 hours. Extraction of total RNA, reverse transcription, PCR amplification and real-time quantitative PCR to observe TLR9 mRNA DSPP mRNA expression levels change. 1% agarose gel electrophoresis, and real-time quantitative PCR results show that CpG ODN-A stimulation group TLR9 mRNA, DSPP mRNA expression levels no significant change. CpG ODN-B stimulation group TLR9 mRNA DSPP mRNA expression levels presented the changes first and then decreased, the highest expression levels of six hours, and a group of six hours 0 hours three times. 3 to participate in the regulation of the expression of the DSPP gene CpG OND three cell signaling pathways within the cell signaling pathway inhibitors: ERK signaling pathway inhibitor, PD98059; p38 signaling pathway inhibitor, SB203580; NF-κB signaling pathway inhibitor, PDTC. OLC cells were treated with the above three inhibitors (concentration of 30μmol / L) after adding CpG ODN-B stimulating six hours after extraction of total RNA, quantitative real-time PCR results showed that ERK and NF-κB group DSPP mRNA expression was significantly reduced; p38 DSPP mRNA expression level was not statistically significant.

Role of Toll Like Receptor 9 Signalling Pathway in Odontoblasts

Abstract

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