人鼻黏膜上皮细胞中白细胞介素12和细胞间黏附分子1及单核细胞趋化因子3的表达研究

Characteristics of mRNA level of interleukin 12 and intercellular adhesion molecule 1 and monocyte chemotactic protein 3 in human nasal epithelial cells

目的:利用RT-PCR检测原代培养的人鼻黏膜上皮细胞中IL-12、细胞间黏附分子1(ICAM-1)以及单核细胞趋化因子3(MCP-3)mRNA的表达。方法:获取人鼻黏膜组织,原代培养人鼻黏膜上皮细胞。设计IL-12 p35亚基、IL-12 p40亚基I、CAM-1以及MCP-3的引物,参照物采用3-磷酸甘油醛脱氢酶(GAPDH),目标扩增片段938 bp。半定量RT-PCR检测其mRNA在鼻黏膜上皮细胞的表达。结果:×400光镜下观察结果显示原代培养的鼻黏膜上皮细胞形状呈圆形或不规则形,饱满贴壁。RT-PCR结果通过1%琼脂糖凝胶电泳显示,鼻黏膜上皮细胞有IL-12 p35I、CAM-1以及MCP-3 mRNA的表达,但无IL-12 p40亚基的表达。结论:人鼻黏膜上皮细胞中有IL-12 p35、ICAM-1以及MCP-3的mRNA表达,但无IL-12p40亚基的表达。人鼻黏膜上皮细胞不能合成完整有活性的IL-12。

Objective：To investigate the mRNA level of IL-12, ICAM-I and MCP-3 in human nasal epithelial cells. Method：Firstly, human primary nasal epithelial cells were cultured, and then 4 pairs of primers were designed for detecting mRNA level of IL-12, ICAM-1 and MCP3. The 938 bp PCR products of GAPDH were used as internal standards. The mRNA expression levels of IL-12, ICAM 1 and MCP-3 in primary nasal epithelial cells was measured with semi quantitative reverse transeription-polymerase chain reaction. Result：. The round or irregu lar primary nasal epithelial cells were observed sticking to the bottom of cell culture plates under 400 times optical microscope. The expressions of IL-12 p35, ICAM-I and MCP-3 mRNA were found in primary nasal epithelial cells while IL 12 p40 subunit was not detected . Conclusion：IL-12 p35, ICAM-I and MCP-3 mRNAs are expressed in primary nasal epithelial cells, whereas effective IL-12 with integrity is not present in nasal epithelial cells.