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.