内毒素耐受对THP-1细胞分泌ENA-78和EGF的影响

Effects of endotoxin tolerance on the production of ENA-78 and EGF in THP-1 cells

目的:观察脂多糖(LPS)诱导的内毒素耐受对THP-1细胞分泌上皮来源的中性粒细胞活化肽78(ENA-78)和表皮生长因子(EGF)的影响。方法:采用1μg/m L牙龈卟啉单胞菌(P.gingivalis)LPS或大肠杆菌(E. coli)LPS分别刺激THP-1细胞24 h,洗脱后,再次刺激24 h,诱导细胞产生耐受。采用ELISA技术检测细胞上清液中ENA-78和EGF表达水平的变化。结果:P.gingivalis LPS单次刺激后,ENA-78水平与空白对照组比较无明显差别(P>0.05),E.coli LPS单次刺激后,ENA-78水平较空白对照组比较明显增高(P<0.05);两种LPS重复刺激后,ENA-78水平均较单次刺激后明显降低(P<0.05)。但两种LPS单次/重复刺激后,EGF分泌水平均无明显变化(P>0.05)。结论:内毒素耐受能抑制THP-1细胞分泌ENA-78,进而可能对中性粒细胞趋化产生影响。

Objective： To observe the effects of endotoxin tolerance on the production of chemokine epithelial neutrophil-activating peptide 78（ ENA-78） and epidermal growth factor（EGF） in THP-1 cells. Methods： THP-1 cells were pretreated with 1 μg/mL P.gingivalis（LPS） or 1 μg/mL E. coli LPS for 24 h,washed and stimulated with the same LPS again for another 24 h. Levels of ENA-78 and EGF in supernatants were detected by ELISA. Results： There were no differences in concentration of ENA-78 secreted by THP-1 cells stimulated with P. gingivalis LPS（P〉0.05）,while the amounts of ENA-78 in the cells challenged by E. coli LPS increased significantly compared with those without any stimulations（P〈0.05）. In addition,secretions of ENA-78 decreased significantly after repeated P. gingivalis LPS or E. coli LPS stimulations compared with those following signal challenge（P〈0.05）. However,no significant changes of EGF levels were detected in THP-1 cells restimulated with P. gingivalis LPS or E. coli LPS compared with those challenged only once（P〉0.05）. Conclusions： Secretions of ENA-78 in THP-1 cells could be suppressed by endotoxin tolerance,which might have an effect on neutrophil migration.