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目的探讨汉滩病毒感染血管内皮细胞后,Toll样受体(TLRs)分子的表达变化,为抗感染免疫和致病机制研究提供重要资料。方法分别用LPS,CL097,PolyI:C以及汉滩病毒76-118刺激血管内皮细胞,6h后提取总RNA,进行反转录获得cDNA,再分别用各自的引物进行PCR反应,产物用1%琼脂糖凝胶电泳分析,并用间接免疫荧光测定汉滩病毒感染血管内皮细胞后TLR4的表达。结果汉滩病毒刺激感染后,血管内皮细胞TLR2、TLR4转录水平增高,TLR3转录水平降低,细胞膜表面的TLR4表达上调。结论汉滩病毒76-118感染血管内皮细胞后,可以引起TLRs分子转录水平发生改变,其中TLR4表达上调,固有免疫参与了汉滩病毒致病过程。
Objective To investigate the expression changes of Toll-like receptors (TLRs) after Hantaan virus infected vascular endothelial cells, and provide important information for anti-infection immunity and pathogenesis. Methods Vascular endothelial cells were stimulated with LPS, CL097, PolyI: C and Hantaan virus 76-118 respectively. After 6 hours, total RNA was extracted and reverse transcribed to obtain cDNA. PCR was carried out with their respective primers. The products were stained with 1% agar The expression of TLR4 in Hantaan virus infected vascular endothelial cells was detected by the method of sugar gel electrophoresis and indirect immunofluorescence. Results After Hantaan virus infection, the transcripts of TLR2 and TLR4 in vascular endothelial cells increased, the transcription level of TLR3 decreased, and the expression of TLR4 on the cell membrane surface was up-regulated. Conclusion The Hantaan virus 76-118 infected vascular endothelial cells can cause changes in TLRs molecular transcription, including TLR4 expression, innate immunity involved in the pathogenesis of Hantaan virus.