Toll样受体4在人成牙本质细胞和牙髓组织中的表达  

The expression of toll like receptor 4 in human odontoblasts and dental pulp tissue

作　　者：蒋宏伟[1] 凌均棨[1] 曾劲峰[1] 

机构地区：[1]中山大学光华口腔医学院.附属口腔医院.口腔医学研究所,广州510055

出　　处：《中华口腔医学研究杂志（电子版）》2008年第1期4-7,共4页Chinese Journal of Stomatological Research(Electronic Edition)

基　　金：国家自然科学基金(30672318);广州市科技局科技攻关计划重点项目(2004Z2-E0021)

摘　　要：目的探讨人牙髓组织和成牙本质细胞中Toll样受体4(TLR4)的表达与分布特点,及其在牙髓组织天然免疫防御中的可能作用。方法从新鲜拔除的人第三磨牙获取牙髓组织和成牙本质细胞,扫描电镜观察牙髓摘除后髓腔壁成牙本质细胞形态和附着情况;RT-PCR检测人牙髓组织以及成牙本质细胞中TLR4mRNA的表达情况;采用免疫印迹技术和免疫组织化学方法检测TLR4蛋白的表达与定位。结果扫描电镜观察牙髓摘除后成牙本质细胞大部存留在髓室壁上,细胞形态良好;RT-PCR结果发现健康牙髓、成牙本质细胞均可表达TLR4mRNA,产物大小为278bp;免疫印迹法检测正常牙髓、成牙本质细胞在相对分子质量89000附近出现蛋白条带;免疫组化显示牙髓组织中的成牙本质细胞、血管内皮细胞TLR4免疫反应阳性。正常牙髓中牙髓成纤维细胞不表达TLR4。结论成牙本质细胞通过表达TLR4参与牙髓牙本质复合体的先天免疫反应,TLR4在牙髓抵御外来病原微生物入侵的宿主免疫防御中起重要作用。Objective To investigate the expression and distribution of Toll like receptor 4 （TLR4） in human dental pulp tissue and odontoblasts and explore the role of TLR4 in the innate immune response. Methods Odontoblasts and pulp tissue were collected from extracted human teeth. Scanning electron microscopy （SEM） was performed on representative teeth to demonstrate the existence of an odontoblast monolayer. TLR4 mRNA levels of human pulp tissue and odontoblasts were assayed by reverse transcription-polymerse chain reaction （RT-PCR）.The expression and localization of TLR4 protein were studied with western blot and immunohistochemistry. Results SEM revealed the integrity of the odontoblast layer and the well-preserved morphology of the individual cells. RT-PCR demonstrated TLR4 mRNA expression in odontoblasts and healthy pulp tissue with a 278bp transcript. Western blot with TLR4-specific antibody also detected the 89 000 protein in these samples. Immunohistochemical staining detected TLR4 in odontoblasts in the dental pulp. Negative immunostaining was observed in the pulp fibroblasts. Some pulp capillary endothelial cell exhibited intense staining. Conclusions Human odontoblasts and pulp tissue express TLR4, which suggests that odontoblasts take part in the innate immune systerm and TLR4 may play an important role in the host defense of human dental pulp.

关 键 词：牙髓 成牙本质细胞 TOLL样受体4 

分 类 号：R78[医药卫生—口腔医学]