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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 detect the expression of TLR9 in primary cultured human nasal epithelial cells by RT-PCR, immunohistochemistry and flow cytometry. Methods: Primary human nasal mucosa epithelial cells were cultured and primers of TLR9 were designed. The expression of TLR9 mRNA in nasal mucosa epithelial cells was detected by RT-PCR. Immunohistochemistry and flow cytometry were used to detect the expression of TLR9 in primary cultured human nasal epithelial cells. Results: The results of 400-fold light microscopy showed that the primary cultured nasal mucosal epithelial cells were round or irregular in shape and full of adherent wall. RT-PCR results showed that the expression of TLR9 mRNA in nasal mucosa epithelial cells. Compared with GAPDH, an internal reference, the expression of TLR9 in nasal mucosa epithelial cells was higher than that in peripheral blood mononuclear cell positive control group, the difference was statistically significant. Conclusion: The expression of TLR9 in human nasal mucosa epithelial cells is higher than that in peripheral blood mononuclear cells in human nasal mucosa epithelial cells.