上海口腔医学

上海口腔医学 ›› 2019, Vol. 28 ›› Issue (2): 128-132.doi: 10.19439/j.sjos.2019.02.004

• 论著 • 上一篇    下一篇

白藜芦醇对牙髓卟啉单胞菌脂多糖诱导成骨细胞产生MIP-2的影响

于雅琼, 李晓琳, 仇丽鸿, 曲柳, 杨谛, 王慧君   

  1. 中国医科大学口腔医学院 牙体牙髓病科,辽宁省口腔医学研究所 牙体牙髓病学研究室, 辽宁省口腔疾病转化医学研究中心,辽宁 沈阳 110002
  • 收稿日期:2018-06-28 修回日期:2018-09-11 出版日期:2019-04-25 发布日期:2019-06-20
  • 通讯作者: 仇丽鸿,E-mail：drqlh@yahoo.com
  • 作者简介:于雅琼（1987-）,女,医师,讲师,E-mail：yuyaqiong1987@126.com
  • 基金资助:
    辽宁省高等学校基本科研项目（青年项目）（LQNK201721）; 中国医科大学口腔医学院青年科研启动基金项目（K101593-16-01）

Effect of resveratrol on the expression of MIP-2 in P.e-LPS-induced mouse osteoblasts

YU Ya-qiong, LI Xiao-lin, QIU Li-hong, QU Liu, YANG Di, WANG Hui-Jun   

  1. Department of Endodontics, School of Stomatology, China Medical University; Lab of Endodontics, Liaoning Institute of Dental Research; Liaoning Provincial Research Center of Translational Oral Medicine. Shenyang 110002, Liaoning Province, China
  • Received:2018-06-28 Revised:2018-09-11 Online:2019-04-25 Published:2019-06-20

摘要： 目的 研究牙髓卟啉单胞菌（Porphyromonas endodontalis,P.e）脂多糖（lipopolysaccharide,LPS）影响成骨细胞产生巨噬细胞炎性蛋白2（macrophage inflammatory protein-2,MIP-2）mRNA和蛋白分泌的作用,以及白藜芦醇对P.e-LPS诱导分泌MIP-2产生的抑制作用。方法 以不同剂量的P.e-LPS (0~50 mg/L)作用于MC3T3-El细胞,并以20 mg/L P.e-LPS作用MC3T3-El细胞不同时间(0~48 h),采用实时反转录PCR(real-time RT-PCR)和酶联免疫吸附试验（ELISA）检测MIP-2 mRNA和蛋白的表达;采用ELISA方法检测白藜芦醇对20 mg/L P.e-LPS 刺激MC3T3-El细胞24 h后MIP-2 蛋白表达的影响。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 当不同剂量P.e-LPS (0~50 mg/L) 刺激MC3T3-El细胞时,MIP-2 mRNA 的表达和蛋白的分泌随剂量的提高而升高;其中,蛋白表达从（41.86±2.49）ng/L升高到（3126.74±158.30）ng/L,差异显著（P<0.05）。在观察时间内（0~48 h）,20 mg/L P.e-LPS刺激MC3T3-El细胞后,MIP-2 mRNA 的表达和蛋白的分泌随时间的延长而增高,48 h时蛋白表达量最高,为（2102.55±123.27）ng/L（P<0.01）。10 mol/L 白藜芦醇预处理细胞1 h,可抑制P.e-LPS诱导的MIP-2 蛋白的表达,蛋白水平从（1805.33±67.54）ng/L降低到（813.82±47.21）ng/L,差异显著（P<0.01）。结论 P.e-LPS 可诱导成骨细胞表达和分泌MIP-2,白藜芦醇对P.e-LPS诱导的MIP-2蛋白分泌发挥抑制作用。

关键词: 牙髓卟啉单胞菌, 脂多糖, 成骨细胞, 巨噬细胞炎性蛋白2, 白藜芦醇

Abstract: PURPOSE: To investigate the effect of lipopolysaccharides(LPS) extracted from Porphyromonas endodontalis(P.e) on the expression of macrophage inflammatory protein-2 (MIP-2) mRNA and protein levels in MC3T3-E1 cells and the influence of resveratrol on the expression of MIP-2 protein in P.e-LPS induced cells. 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 (0-48 h). The expression of MIP-2 mRNA and protein was detected by real-time RT-PCR and enzyme linked immunosorbent assay (ELISA). MC3T3-E1 cells were pretreated with resveratrol for 1 h in the presence of 20 mg/L P.e-LPS for 24 h,which was detected by ELISA. Statistical analysis was performed using one-way ANOVA and Dunnett t test with SPSS 13.0 software package. RESULTS: Treatment of MC3T3-El cells with different concentrations of P.e-LPS(0-50 mg/L) caused a significantly increase in MIP-2 mRNA and protein expression in dose-dependent manners.The expression of MIP-2 protein increased from (41.86±2.49) ng/L to (3126.74±158.30) ng/L, and the difference was significant(P<0.05). In the observation time (0-48 h), the impact of 20 mg/L P.e-LPS on induction of MIP-2 in MC3T3-El cells exhibited a time-dependent manner. At 48 h, the maximal induction of MIP-2 protein expression was (2102.55±123.27) ng/L(P<0.01). Incubation of cells with 10 μmol/L resveratrol for 1h significantly decreased the expression of MIP-2 protein from (1805.33±67.54) ng/L to（813.82±47.21) ng/L, and the difference was significant(P<0.05). CONCLUSIONS: The results suggest that P.e-LPS may mediate MIP-2 expression in MC3T3-E1 cells, and resveratrol has a significant inhibitory effect on this process.

Key words: Porphyromonas endodontalis, Lipopolysaeeharides, Osteoblast, Macrophage inflammatory protein-2, Resveratrol

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