牙髓卟啉单胞菌脂多糖对小鼠成骨细胞表达IL-34 mRNA的影响

doi:10.19439/j.sjos.2017.01.008

摘要/Abstract

摘要： 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）mRNA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 mRNA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 mRNA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 mRNA 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 mRNA 的表达量最大;48 h 时,IL-34 mRNA 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 mRNA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 mRNA,而10 mol/L EX-527则上调IL-34 mRNA的表达。结论 P.e-LPS 可诱导成骨细胞表达IL-34 mRNA,其机制可能是激活p38MAPK、ERK1/2、NF-κB和SIRT1信号通路。

关键词: 牙髓卟啉单胞菌, 脂多糖, 成骨细胞, 白介素34, 信号通路

Abstract: PURPOSE: To investigate the effects of lipopolysaccharides (LPS) extracted from Porphyromonas endodontalis (P.e) on the expression of interleukin-34 (IL-34) mRNA in MC3T3-E1 cells and the role of p38MAPK, ERK1/2, NF-κB and SIRT1 in the process. METHODS: MC3T3-E1 cells were treated with different concentrations of P.e-LPS(0-50 mg/L) and 20 mg/L P.e-LPS for different time (0-24 h). The expression of IL-34 mRNA was detected by real-time reverse transcription-polymerase chain reaction (real time RT-PCR). MC3T3-E1 cells were pretreated with inhibitor of NF-κB(BAY 11-7082),inhibitor of p38MAPK (SB203580), inhibitor of ERK1/2 (PD98059), agonist of sirtuin1 (SIRT1) [resveratrol (RES)] and inhibitor of SIRT1 (EX-527) for 1 h, and then were treated with 20 mg/L P.e-LPS. The expression of IL-34 mRNA was detected by real time RT-PCR. Statistical analysis was performed using one-way ANOVA and Dunnett t test with SPSS 13.0 software package. RESULTS: The level of IL-34 mRNA increased significantly after treatment with different concentrations of P.e-LPS(0-50 mg/L),which indicated that P.e-LPS induced osteoblasts to express IL-34 mRNA in a dose-dependent manner. Maximal induction of IL-34 mRNA expression was observed in MC3T3-E1 cells treated with 20 mg/L P.e-LPS for 24 h.At 48 h, the expression of IL-34 mRNA decreased gradually. The mRNA of IL-34 decreased significantly after pretreatment with 10 μmol/L BAY-117082, SB203580 and PD98059 for 1 h. P.e-LPS-induced IL-34 upregulation was attenuated by pretreatment with RES, but increased by EX-527. CONCLUSIONS: These results suggest that P.e-LPS may mediate IL-34 mRNA expression in MC3T3-E1 cells. This process is dependent, at least in part, on p38MAPK, ERK1/2, NF-κB and SIRT1 signaling pathways.

Key words: Porphyromonas endodontalis, Lipopolysaeeharides, Osteoblast, Interleukin-34, Signaling pathways

中图分类号:

R780.2

于雅琼, 郭佳杰, 仇丽鸿, 李晓琳, 杨谛, 郭艳. 牙髓卟啉单胞菌脂多糖对小鼠成骨细胞表达IL-34 mRNA的影响[J]. 上海口腔医学, 2017, 26(1): 37-41.

YU Ya-qiong, GUO Jia-jie, QIU Li-hong, LI Xiao-lin, YANG Di, GUO Yan. Effect of lipopolysaccharides from Porphyromonas endodontalis on the expression of interleukin-34 in mouse osteoblasts[J]. Shanghai Journal of Stomatology, 2017, 26(1): 37-41.

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