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目的 :探讨牙髓卟啉单胞菌(Porphyromonas endodontalis,P.e)脂多糖(lipopolysaccharide,LPS)对成骨细胞产生白介素34(interleukin-34,IL-34)m RNA表达的影响及此过程是否有p38MAPK、ERK1/2、NF-κB和SIRT1蛋白的参与。方法:以不同浓度P.e-LPS(0~50 mg/L)刺激MC3T3-El细胞和以20 mg/L P.e-LPS作用细胞不同时间(0~24 h)后,采用实时反转录聚合酶链反应(real-time RT-PCR)检测IL-34 m RNA的表达。以同样方法检测NF-κB抑制剂BAY 11-7082、p38MAPK抑制剂SB203580、ERK1/2抑制剂PD98059、沉默调节蛋白1(sirtuin1,SIRT1)激动剂白藜芦醇(resveratrol,RES)和SIRT1抑制剂EX-527对P.e-LPS刺激MC3T3-El细胞后IL-34 m RNA表达的影响。采用SPSS 13.0软件包对结果进行单因素方差分析和Dunnett t检验。结果 :不同浓度P.e-LPS(0~50 mg/L)刺激MC3T3-El细胞后,IL-34 m RNA的表达具有剂量依赖性。20 mg/L P.e-LPS作用MC3T3-El细胞24 h时,IL-34 m RNA的表达量最大;48 h时,IL-34 m RNA的表达量有所下降。10 mol/L BAY-117082、SB203580、PD98059预处理细胞1 h,可以降低P.e-LPS诱导的IL-34 m RNA的表达水平。50 mol/L RES下调P.e-LPS诱导小鼠成骨细胞表达IL-34 m RNA,而10 mol/L EX-527则上调IL-34 m RNA的表达。结论:P.e-LPS可诱导成骨细胞表达IL-34 m RNA,其机制可能是激活p38MAPK、ERK1/2、NF-κB和SIRT1信号通路。
Objective: To investigate the effects of Porphyromonas endodontalis (Pe) lipopolysaccharide (LPS) on the expression of interleukin-34 (IL-34) m RNA in osteoblasts and whether p38MAPK , ERK1 / 2, NF-κB and SIRT1 proteins. Methods: MC3T3-E1 cells were treated with different concentrations of Pe-LPS (0-50 mg / L) and cells were treated with 20 mg / L Pe-LPS for different time (0-24 h). Real-time reverse transcription polymerase chain The expression of IL-34 mRNA was detected by real-time RT-PCR. In the same way, NF-κB inhibitors BAY 11-7082, p38MAPK inhibitor SB203580, ERK1 / 2 inhibitor PD98059, sirtuin1 (SIRT1) agonist resveratrol (RES) and SIRT1 inhibitor Effect of EX-527 on the expression of IL-34 mRNA after Pe-LPS stimulation of MC3T3-El cells. One-way ANOVA and Dunnett t-test were used to analyze the results using SPSS 13.0 software package. Results: The expression of IL-34 mRNA in MC3T3-El cells stimulated with different concentration of P.e-LPS (0-50 mg / L) was dose-dependent. At 20 mg / L P.e-LPS, the expression of IL-34 mRNA was the highest at 24 h in MC3T3-El cells, and decreased at 48 h. Pretreatment of cells with 10 mol / L BAY-117082, SB203580 and PD98059 for 1 h reduced P.e-LPS-induced IL-34 mRNA expression. 50 mol / L RES down-regulated the expression of IL-34 mRNA in P.e-LPS-induced mouse osteoblasts, while 10 mol / L EX-527 up-regulated the expression of IL-34 mRNA. CONCLUSION: P.e-LPS can induce the expression of IL-34 mRNA in osteoblasts, which may be caused by the activation of p38MAPK, ERK1 / 2, NF-κB and SIRT1 signaling pathways.