Yu Ya-Qiong, Guo Jia-Jie, Qiu Li-Hong, Li Xiao-Lin, Yang Di, Guo Yan
Department of Endodontics, School of Stomatology, China Medical University. Shenyang 110002,Liaoning Province, China. E-mail:
Shanghai Kou Qiang Yi Xue. 2017 Feb;26(1):37-41.
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.
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.
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.
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.
研究从牙髓卟啉单胞菌（P.e）中提取的脂多糖（LPS）对MC3T3-E1细胞中白细胞介素-34（IL-34）mRNA表达的影响，以及p38丝裂原活化蛋白激酶（p38MAPK）、细胞外信号调节激酶1/2（ERK1/2）、核因子κB（NF-κB）和沉默信息调节因子1（SIRT1）在此过程中的作用。
用不同浓度的P.e-LPS（0 - 50 mg/L）和20 mg/L P.e-LPS处理MC3T3-E1细胞不同时间（0 - 24 h）。通过实时逆转录-聚合酶链反应（实时RT-PCR）检测IL-34 mRNA的表达。MC3T3-E1细胞先用NF-κB抑制剂（BAY 11-7082）、p38MAPK抑制剂（SB203580）、ERK1/2抑制剂（PD98059）、沉默信息调节因子1（SIRT1）激动剂[白藜芦醇（RES）]和SIRT1抑制剂（EX-527）预处理1 h，然后用20 mg/L P.e-LPS处理。通过实时RT-PCR检测IL-34 mRNA的表达。使用SPSS 13.0软件包进行单因素方差分析和Dunnett t检验进行统计分析。
用不同浓度的P.e-LPS（0 - 50 mg/L）处理后，IL-34 mRNA水平显著升高，这表明P.e-LPS以剂量依赖性方式诱导成骨细胞表达IL-34 mRNA。在用20 mg/L P.e-LPS处理24 h的MC3T3-E1细胞中观察到IL-34 mRNA表达的最大诱导。在48 h时，IL-34 mRNA的表达逐渐下降。用10 μmol/L BAY-117082、SB203580和PD98059预处理1 h后，IL-34的mRNA显著下降。RES预处理减弱了P.e-LPS诱导的IL-34上调，但EX-527预处理则使其增加。
这些结果表明，P.e-LPS可能介导MC3T3-E1细胞中IL-34 mRNA的表达。这一过程至少部分依赖于p38MAPK、ERK1/2、NF-κB和SIRT1信号通路。
