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目的:探讨核因子κB(NF-κB)在牙髓卟啉单胞菌(P.e)脂多糖(LPS)诱导小鼠成骨细胞株MC3T3-El白介素6(IL-6)表达中的作用。方法:10 mg/L P.e-LPS刺激MC3T3-El细胞不同时间后,应用免疫荧光方法检测NF-κB核易位情况和BAY-117082对NF-κB核易位抑制情况,反转录聚合酶链反应(RT—PCR)和酶联免疫吸附试验(ELISA)检测BAY-117082对MC3T3-El细胞的IL-6基因和蛋白表达的影响。采用SPSS 13.0软件包对结果进行多因素方差分析和Dunnett t检验。结果:在静息状态下,NF-κB定位在胞质中,10 mg/L P.e-LPS刺激MC3T3-El细胞30 min后即引起NF-κB快速的核易位;60 min后,大部分NF-κB重新定位在胞质里,10μmol/L BAY-117082预处理1 h可抑制P.e-LPS引起的NF-κB核易位,降低P.e-LPS诱导MC3T3-El细胞IL-6 mRNA表达水平和IL-6蛋白的分泌水平,其中,蛋白分泌水平从(774.983±6.585)ng/L降低至(377.384±14.620)ng/L(P<0.O1),而BAY-117082单独刺激组与空白对照组无显著差异。结论:P.e-LPS可诱导MC3T3-El细胞中NF-κB的核易位,P.e-LPS可通过激活NF-κB信号途径诱导MC3T3-El细胞产生IL-6。
AIM: To investigate the role of nuclear factor-κB (NF-κB) in the expression of interleukin-6 (IL-6) in mouse osteoblastic cell line MC3T3-El induced by PPS lipopolysaccharide (LPS). Methods: After MC3T3-E1 cells were treated with 10 mg / L Pe-LPS for different time, the nuclear translocation of NF-κB and the inhibition of nuclear translocation of NF-κB by BAY-117082 were detected by immunofluorescence. The effect of BAY-117082 on the expression of IL-6 gene and protein in MC3T3-El cells was detected by RT-PCR and enzyme-linked immunosorbent assay (ELISA). The results were analyzed by multivariate analysis of variance and Dunnett t test using SPSS 13.0 software package. Results: At resting state, NF-κB was localized in the cytoplasm. After MC3T3-E1 cells were treated with 10 mg / L Pe-LPS for 30 min, nuclear translocation of NF-κB was rapidly induced. After 60 min, most NF The pretreatment with 10μmol / L BAY-117082 for 1 h inhibited the nuclear translocation of NF-κB induced by Pe-LPS and the IL-6 mRNA expression and IL-6 in MC3T3-El cells induced by Pe-LPS The level of protein secretion decreased from (774.983 ± 6.585) ng / L to (377.384 ± 14.620) ng / L (P <0.01), while the BAY-117082 alone stimulation group No significant difference. CONCLUSION: P.e-LPS can induce nuclear translocation of NF-κB in MC3T3-El cells. P.e-LPS can induce IL-6 production in MC3T3-El cells by activating NF-κB signaling pathway.