脂多糖刺激树突状细胞前后TLR2、3、4基因表达的检测

Detection of Gene Expression for TLR2,3,4 on Dendritic Cells before and after Stimulation by Lipopolysaccharide

目的检测脂多糖作用于树突状细胞前后TLR2、3、4基因的表达情况。方法从外周血分离单核细胞,加入rhGMCSF及rhIL4,培养的第6天,实验组加入脂多糖刺激24h,对照组不加。分别提取细胞总RNA,行半定量RTPCR,产物进行琼脂糖凝胶电泳分析。结果单核细胞经GMCSF及IL-4诱导7d后的DC表达较高水平TLR2、3、4。脂多糖刺激24h后,TLR2、3、4表达下降,TLR4几乎测不到。结论树突状细胞在受脂多糖刺激前后TLRs的表达有显著变化,推测和它的功能成熟有关。

Objective To detect the gene expression for TLR2,3,4 on dendritic cells before and after stimulation by lipopolysaccharide. Method Monocytes were isolated from heparinized whole blood. 7 d later the monocytes were induced into dendritic cells by adding rhGM-CSF and rhIL-4. Total cellular RNA of unstimulated and stimulated DC were extracted by LPS 24 h later. Semi-quantitative RT-PCR,agarose gel electrophoresis were conducted to analyse the PCR products. Result Expression of TLR2,3,4 were significantly higher in monocytes-derived DC induced to 7 d.After being stimulated by LPS for 24 h ,the expression of TLR2,3,4 decreased, and that of TLR4 was almost undetectable. Conclusion The expression of TLR2,3,4 on DC changes significantly after being stimulated by LPS.This may be related to functional maturation of DC.