TLR_4在人牙周膜成纤维细胞中的表达研究

Study on the Expression of TLR_4 in Human Periodontal Ligament Fibroblasts

目的:探讨TLR4在体外培养的人牙周膜成纤维细胞中的表达情况。方法:体外分离培养人牙周膜成纤维细胞,采用免疫荧光染色检测TLR4蛋白在该细胞中的表达;用半定量RT-PCR法检测TLR4基因的表达及0.1 ng/mL牙龈卟啉菌脂多糖刺激24 h后细胞内TLR4基因表达水平的改变。结果:免疫荧光检测显示体外培养的人牙周膜细胞中有TLR4表达,0.1 ng/mL牙龈卟啉菌脂多糖刺激24 h,其TLR4基因表达水平显著性增高(P<0.05)。结论:TLR4在牙周膜成纤维细胞中有表达,牙龈卟啉菌脂多糖成分可刺激TLR4基因上调,提示与该细胞参与牙周免疫应答有关。

Objective： To investigate the expression of TLR4 in human periodontal ligament fibroblasts and the effect of lipopolysaccharide of porphyromonas gingivalis （Pg. LPS） on the expression of TLR4. Methods： Periodontal fibroblasts come from healthy human premolars extracted due to orthodontic treatment. The 3rd or 4th passage cells were prepared for test. Immunofluoresence technique was used to detect the expression of TLR4 on the periodontal ligament fibroblasts. 0. 1 ng/ml Pg. LPS was added to the culture media, and RT - PCR was used to measure the level of TLR4 expression 24h later. Results： It was found that there were TLR4 expressed in the human periodontal ligament fibroblasts. And gene expression of TLR4 of the cells stimulated with 0.1 ng/ml Pg. LPS was higher than that of non - stimulated cells. The difference was statistically significant （ P 〈 0.05 ）. Conclusion ： There are TLR4 expressed in periodontal ligament fibroblasts. Pg. LtX3 can up - regulate the level of TLR4 gene expression. This means that TLR4 may play roles in periodontal immunity reaction.