喜树碱对NIH／3T3细胞TLR3表达及活化的影响

Effect of CPT on TLR3 expression and activation in NIH/3T3 cell

为研究喜树碱(CPT)对小鼠成纤维细胞NIH/3T3中Toll样受体3(TLR3)的表达及活化的影响,本实验利用CCK-8试剂盒测定了CPT在不同浓度下、不同作用时间对NIH/3T3细胞的毒性作用,并确定CPT对NIH/3T3细胞的基本无毒浓度为1μg/mL。通过流式细胞术检测CPT作用NIH/3T3细胞表明,TLR3平均荧光强度与对照组相比显著增强,表明TLR3表达增加。此外,ELISA试剂盒检测结果显示CPT作用后IFN-β表达量增加,并高于阳性对照组,差异极显著(p<0.01)。本研究表明CPT对NIH/3T3细胞的TLR3具有显著的激动作用。

To study the effect of CPT on TLR3 expression and activation in mouse fibroblast NIH/3T3 cell. CCK-8 was used to test toxicity of CPT to NIH/3T3 cell at different concentration and different time, and 1 ug/mL of CPT was determined as the basic non-toxic concentration. The FACS result indicated that, compared with negative control group, the statistical of mean fluorescence intensity that cell co-culture with CPT is significantly enhanced after the cell co-cultured with CPT. It showed that the expression of TLR3 was increased in the cells. Furthermore, compared with positive control, expression of IFN-β was increased significantly （p〈0.01） after co-culture with CPT detected by ELISA kit. The present study demonstrated that CPT was able to be as TLR3 agonist in NIH/3T3 cell.