牙髓卟啉单胞菌脂多糖对小鼠成骨细胞表达MCP-1的影响

doi:10.19439/j.sjos.2018.01.001

上海口腔医学 ›› 2018, Vol. 27 ›› Issue (1): 1-5.doi: 10.19439/j.sjos.2018.01.001

牙髓卟啉单胞菌脂多糖对小鼠成骨细胞表达MCP-1的影响

李晓琳^1,2, 于雅琼^1,2, 仇丽鸿^1,2, 郭佳杰^1,2, 邵丽娜^1,2, 王丝墨^1,2, 杨谛^1,2

1.中国医科大学口腔医学院牙体牙髓病学教研室,辽宁沈阳 110002
2.辽宁省口腔医学研究所牙体牙髓病学研究室,辽宁沈阳 110002

收稿日期:2016-12-13 修回日期:2017-04-04 出版日期:2018-02-25 发布日期:2018-03-05
通讯作者: 仇丽鸿,E-mail: drqlh@yahoo.com
作者简介:李晓琳（1991-）,女,医师,E-mail: aboutlxl@sina.com
基金资助:
国家自然科学基金（81500843）; 国家人力资源与社会保障部留学人员科技活动项目（2016-B）; 沈阳市科技计划项目（F15-199-1-56）

Effects of Porphyromonas endodontalis lipopolysaccharides on the expression of monocyte chemotactic protein-1 in mouse osteoblasts

LI Xiao-lin^1,2, YU Ya-qiong^1,2, QIU Li-hong^1,2, GUO Jia-jie^1,2, SHAO Li-na^1,2, WANG Si-mo^1,2, YANG Di^1,2

1.Department of Endodontics, School of Stomatology, China Medical University. Shenyang 110002
2.Lab of Endodontic, Liaoning Institute of Dental Research. Shenyang 110002.Liaoning Province, China

Received:2016-12-13 Revised:2017-04-04 Online:2018-02-25 Published:2018-03-05

摘要/Abstract

摘要： 目的: 探讨牙髓卟啉单胞菌(Porphyromonas endodontalis,P.endodontalis)脂多糖对小鼠成骨细胞表达单核细胞趋化蛋白1(monocyte chemotactic protein-1, MCP-1)mRNA和蛋白的影响,以及是否有p38丝裂原活化蛋白激酶（mitogen-activated protein kinase,MAPK）信号通路和核因子κB（Nuclear Factor-κB, NF-κB）信号通路的参与。方法: 以不同质量浓度的P.endodontalis脂多糖(0~50 mg/L)刺激MC3T3-E1细胞24 h;以20 mg/L P.endodontalis脂多糖作用于细胞不同时间（0~48 h）后,采用实时反转录PCR和酶联免疫吸附测定检测MCP-1mRNA和蛋白的表达。采用p38MAPK抑制剂SB203580和NF-κB抑制剂BAY11-7082分别预处理细胞1 h,检测其对P.endodontalis脂多糖刺激MC3T3-E1细胞后MCP-1mRNA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: 不同质量浓度的P.endodontalis脂多糖（0~50 mg/L）刺激MC3T3-E1细胞后,MCP-1的mRNA表达和蛋白分泌呈剂量依赖性;观察时间内（0~48 h）,20 mg/L P.endodontalis脂多糖作用于MC3T3-E1细胞后,MCP-1 mRNA的表达和蛋白分泌呈时间依赖性;10 mol/L的SB203580和BAY11-7082分别预处理细胞1 h,可以降低P.endodontalis脂多糖诱导MCP-1 mRNA的表达,且SB203580的抑制作用强于BAY11-7082。结论: P.endodontalis脂多糖可能通过激活p38MAPK和NF-κB信号通路诱导成骨细胞表达MCP-1mRNA和蛋白。

关键词: 牙髓卟啉单胞菌, 脂多糖, 单核细胞趋化蛋白1, 成骨细胞, 信号通路

Abstract: PURPOSE: To investigate the effects of lipopolysaccharide (LPS) extracted from Porphyromonas endodontalis (P.endodontalis) on expression of monocyte chemotactic protein-1 (MCP-1) mRNA and protein in MC3T3-E1 cells and the role of p38 mitogen-activated protein kinase (MAPK) and nuclear factor-κB(NF-κB）in the process. METHODS: MC3T3-E1 cells were treated with different concentrations of P.endodontalis LPS(0-50mg/L) and 20 mg/L P.endodontalis LPS for different hours (0-48 h). The expression of MCP-1 mRNA was detected by real-time reverse transcription PCR(RT-PCR) and protein was detected by enzyme-1inked immunosorbent assay (ELISA). MC3T3-E1 cells were pretreated with SB203580 (inhibitor of p38MAPK) and BAY11-7082 (inhibitor of NF-κB) for 1h, and then were treated with 20 mg/L P.endodontalis LPS for 24 h, the expression of MCP-1 mRNA was also detected by RT-PCR. Statistical analysis was performed using one-way ANOVA and Dunnett t test with SPSS 13.0 software package. RESULTS: The level of MCP-1 mRNA and protein increased significantly after treatment with different concentrations of P.endodontalis LPS (0-50 mg/L), which indicated that P.endodontalis LPS induced osteoblasts to express MCP-1 in a dose dependent manners. During the observation time (0-48 h), the impact of 20 mg/L P.endodontalis LPS on induction of MCP-1 in MC3T3-E1 cells exhibited a time-dependent manner. The expression of MCP-1 mRNA decreased significantly after pretreated with 10 mol/L SB203580 and BAY11-7082 for 1 h,and the inhibitory effect of SB203580 was stronger than BAY11-7082. CONCLUSIONS: P.endodontalis LPS may induce the expression of MCP-1 mRNA and protein in MC3T3-E1 cells through the signaling pathway of p38MAPK and NF-κB.

Key words: Porphyromonas endodontalis, Lipopolysaeeharides, Monocyte chemotactic protein-1, Osteoblast, Signaling pathway

中图分类号:

R780.2

李晓琳, 于雅琼, 仇丽鸿, 郭佳杰, 邵丽娜, 王丝墨, 杨谛. 牙髓卟啉单胞菌脂多糖对小鼠成骨细胞表达MCP-1的影响[J]. 上海口腔医学, 2018, 27(1): 1-5.

LI Xiao-lin, YU Ya-qiong, QIU Li-hong, GUO Jia-jie, SHAO Li-na, WANG Si-mo, YANG Di. Effects of Porphyromonas endodontalis lipopolysaccharides on the expression of monocyte chemotactic protein-1 in mouse osteoblasts[J]. Shanghai Journal of Stomatology, 2018, 27(1): 1-5.

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牙髓卟啉单胞菌脂多糖对小鼠成骨细胞表达MCP-1的影响

Effects of Porphyromonas endodontalis lipopolysaccharides on the expression of monocyte chemotactic protein-1 in mouse osteoblasts

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