Toll样受体-4在人肺腺上皮细胞、单核细胞和脐静脉血管内皮细胞的表达

Human Toll like receptor-4 expression in human pulmonary granule epithelial cells, monocytes and umbilical vein endothelial cells

目的 检测炎症信号Toll样受体 - 4 (TLR 4 )在人肺腺上皮细胞系 (SPC A 1)、单核吞噬细胞系 (THP 1)和脐静脉血管内皮细胞 (HUVECs)的表达情况。方法 设计人TLR 4PCR特异引物 ,从SPC A 1、THP 1和HUVECs提取总RNA ,采用一步法RT PCR和地高辛标记的Northern杂交技术检测TLR 4mRNA在三种细胞的表达 ;用免疫荧光细胞化学染色在蛋白质水平检测TLR 4在三种细胞的表达。结果 RT PCR、Northern杂交和免疫荧光细胞化学染色均显示 ,三种细胞均表达TLR 4受体。结论 本实验结果表明除单核吞噬细胞外 ,大血管内皮细胞和肺腺上皮细胞亦表达Toll受体基因 。

Objective To determine whether human Tolllike receptor-4(TLR-4) are expressed in human pulmonary granule epithelial cells, monocytes and umbilical vein endothelial cells (HUVECs). Methods Specific primer for TLR-4 was designed and total RNAs were isolated from human pulmonary granule epithelial cell line SPC-A-1, human monocyte cell line THP-1 andthe primary culture of human UVECs. RT-PCR and Northern blotting with digoxin labeling were used for detection of TLR-4 mRNA expression. Immunocytofluorescentstaining was used for detection of TLR-4 in the 3 types of cells. Results TLR-4 was expressed in the 3 types of cells detected by RT-PCR, Northern blotting and immunocytofluorescent staining. By using DNA sequencing, the RT-PCR products were confirmed to be TLR-4 cDNAs. Conclusion The results provide an evidence indicating that TLR-4 constitutively expresses not only in human monocytes, but also in human larger vascular endothelial cells and pulmonary granule epithelial cells. It indicats that TLR-4 may mediate transcriptional activation of inflammatory madiator genes in the three cells.