牙髓卟啉单胞菌脂多糖对成骨细胞CD14表达的影响

Effect of lipopolysaccharides extracted from Porphyromonas endodontalis on the expression of CD14 in osteoblast

目的:探讨牙髓卟啉单胞菌(P.e)脂多糖(LPS)对小鼠成骨细胞株MC3T3-E1细胞CD14表达的影响。方法:在有血清和无血清存在的情况下,用10μg/mLP.e-LPS作用于MC3T3-E1细胞24h,流式细胞术检测CD14的表达;10μg/mLP.e-LPS刺激MC3T3-E1细胞后,观察不同时间(0、1、3、6、12及24h)后检测细胞膜结合型CD14mRNA表达的变化。应用SPSS13.0软件包对结果进行两样本t检验、单因素方差分析和Dunnettt检验。结果:流式细胞术分析P.e-LPS作用24h后,CD14蛋白表达量增加,但无血清组增加更明显;随着P.e-LPS作用时间的增加,MC3T3-E1细胞膜结合型CD14mRNA的表达量逐渐增加,作用3h时表达最明显,作用大于6h后表达量逐渐降低。结论:P.e-LPS可诱导成骨细胞MC3T3-E1的CD14表达增强,表明P.e-LPS可能通过CD14对成骨细胞发挥作用。

PURPOSE：To investigate the effect of lipopolysaccharides（LPS） extracted from Porphyromonas endodontalis （P.e） on the expression of CD14 in osteoblast.METHODS：Under the condition with or without serum, MC3T3-E1 cells were stimulated with 10μg/mL P.e-LPS for 24 hours, then the expression of CD14 was measured using flow cytometry; the membrane CD14 mRNA was detected at different time point （0,1,3,6,12,24h） by RT-PCR.Statistical analysis was performed using two-sample t test,one-way ANOVA and Dunnett t test with SPSS13.0 software package.RESULTS：Flow cytometry showed that CD14 protein increased after the stimulation of P.e-LPS for 24h, while the non-serum group demonstrated more increase; the membrane CD14 mRNA level was up-regulated by 10μg/mL P.e-LPS at 1 hour, and reached the maximum at 3h and declined after 6h.CONCLUSIONS：P.e-LPS can induce the expression of CD14 in osteoblast MC3T3-E1, which indicates that P.e-LPS may play an important role in osteoblast through CD14.