TLR9在人鼻黏膜上皮细胞中的表达及意义

Expression of TLR9 in human nasal epithelial cells

目的:利用RT-PCR、免疫组织化学及流式细胞术检测原代培养的人鼻黏膜上皮细胞TLR9的表达水平及意义。方法:原代培养人鼻黏膜上皮细胞,设计TLR9的引物,RT-PCR检测TLR9 mRNA在鼻黏膜上皮细胞中的表达。免疫组织化学及流式细胞术检测原代培养的人鼻黏膜上皮细胞中TLR9的表达水平。结果:400倍光镜下观察结果显示,原代培养的鼻黏膜上皮细胞呈圆形或不规则形,饱满贴壁。RT-PCR结果显示,鼻黏膜上皮细胞有TLR9 mRNA的表达。通过与内参照物GAPDH进行灰度分析比较,发现鼻黏膜上皮细胞中TLR9的表达高于外周血单个核细胞阳性对照组,差异有统计学意义。结论:人鼻黏膜上皮细胞中有TLR9的表达,TLR9 mRNA在人鼻黏膜上皮细胞中的表达高于外周血单个核细胞。

Objective：To analyze expression of TLR9 in human nasal epithelial cells by semi-quantitative reverse transcription-polymerase chain reaction, immunohistochemistry and flow cytometry. Method： Human primary nasal epithelial cells （HNECs）were cultured, and then analyze the expression of TLR9 in primary cultured HNECs measured by semi-quantitative RT-PCR and immunohistochemistry and flow cytometry. Result： Primary cultured HNECs were observed by 400 × optical microscopes. Round or irregular cells stick to the bottom of cell culture plates. RT-PCR showed that mRNA expressions of TLR9 were found in primary nasal epithelial cells by 1 % agarose gel electrophoresis. Interestingly, by analyzing with gray level of GAPDH, expression of TLR9 mRNA in HNECs was higher than positive control PBMCs. Conelusion：TLR9 is expressed by HNECs. Expression of TLR9 mRNA in nasal epithelial cells is higher than in PBMCs.