华西口腔医学杂志

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牙龈卟啉单胞菌脂多糖对人脐静脉内皮细胞表达趋化因子RANTES和分形素的影响

漆晓玲1,2  赵蕾3  陈珊珊1  孟姝3  吴亚菲3   

  1. 1.口腔疾病研究国家重点实验室 华西口腔医院(四川大学),成都 610041;2.重庆医科大学附属口腔医院牙周病科,重庆 401147;3.口腔疾病研究国家重点实验室 华西口腔医院牙周病科(四川大学),成都 610041
  • 收稿日期:2015-09-30 修回日期:2015-11-21 出版日期:2016-04-01 发布日期:2016-04-01
  • 通讯作者: 吴亚菲,教授,博士,E-mail:yafeiwu@tom.com
  • 作者简介:漆晓玲,硕士,E-mail:815655244@qq.com
  • 基金资助:

    国家自然科学基金(30973323,81371150)

Effects of Porphyromonas gingivalis lipopolysaccharide on the expression of RANTES and fractalkine in human umbilical vein endothelial cells

Qi Xiaoling1,2, Zhao Lei3, Chen Shanshan1, Meng Shu3, Wu Yafei3.   

  1. 1. State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China; 2. Dept. of Periodontics, Stomatological Hospital of Chongqing Medical University, Chongqing 401147, China; 3. State Key Laboratory of Oral Diseases, Dept. of Periodontics, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
  • Received:2015-09-30 Revised:2015-11-21 Online:2016-04-01 Published:2016-04-01
  • Contact: Wu Yafei, E-mail: yafeiwu@ tom.com.
  • Supported by:

    Natural Science Foundation of China (30973323, 81371150).

摘要:

目的  观察牙龈卟啉单胞菌脂多糖(Pg-LPS)对人脐静脉内皮细胞(HUVEC)表达调节活化正常T细胞表达和分泌的趋化因子(RANTES)和分形素的影响。方法  应用不同质量浓度(200、500、1 000 ng·mL-1)的Pg-LPS分别处理HUVEC 1、6、12、24 h,利用实时荧光定量聚合酶链反应(RT-PCR)及酶联免疫吸附(ELISA)法检测RANTES及分形素mRNA及蛋白质的表达变化。结果  在Pg-LPS与HUVEC共同培养1、6和12 h时,除培养时间为12 h、Pg-LPS质量浓度为200 ng·mL-1组的RANTES mRNA和1 h、200 ng·mL-1组RANTES蛋白表达与对照组无明显差异外,其余各实验组RANTES蛋白和mRNA表达量及分形素mRNA表达量均高于对照组,差异有统计学意义(P<0.05);6 h时,二者mRNA表达量达到峰值,分别为对照组的4.88倍和6.20倍;刺激6 h后,RANTES蛋白和mRNA表达量及分形素的mRNA表达量均降低,24 h时,仅Pg-LPS质量浓度为500 ng·mL-1组与对照组相比有统计学差异(P<0.05);分形素蛋白的表达量则仅在Pg-LPS浓度为1 000 ng·mL-1刺激6、12 h时与对照组相比有统计学差异(P<0.05)。结论  Pg-LPS感染具有上调HUVEC表达趋化因子RANTES和分形素的作用,可能在牙周炎促进动脉粥样硬化发生、发展的过程中起一定作用。

关键词: 动脉粥样硬化, 牙周炎, 趋化因子, 牙龈卟啉单胞菌, 脂多糖

Abstract:

Objective  A study was conducted to investigate the effects of Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) on the expression of regulated upon activation normal T-cell expressed and secreted (RANTES) and fractalkine in human umbilical vein endothelial cells (HUVECs). Methods  HUVECs were incubated with different concentrations of Pg-LPS (200, 500, and 1 000 ng·mL−1) for 1, 6, 12, and 24 h, respectively. Then real time quantitative polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent method (ELISA) were adopted to detect the protein levels and mRNA levels of RANTES and fractalkine. Results  The RANTES protein levels and mRNA levels, as well as fractalkine mRNA levels, were significantly higher in all experimental groups of 1, 6, and 12 h than in the control group (P<0.05), except the expression of RANTES mRNA in 200 ng·mL−1 group of 12 h and RANTES protein in 200 ng·mL−1 group of 1 h. The expression levels of RANTES mRNA and fractalkine mRNA were highest in 1 000 ng·mL−1 group of 6 h and were 4.88- and 6.20-fold higher, respectively, than those in the control group. The expression levels of RANTES protein, mRNA, and fractalkine mRNA decreased 6 h after stimulation, and were significantly higher than those in the control group (P<0.05) in the 500 ng·mL−1 group of 24 h. There was a significant difference between the expression of fractalkine mRNA in 1 000 ng·mL−1 group of 6 and 12 h than in the control group (P<0.05). Conclusion  Pg-LPS infection might up-regulate the expression of RANTES and fractalkine in HUVEC, and such expression is important in the development of atherosclerosis.

Key words: atherosclerosis, periodontitis, chemokines, Porphyromonas gingivalis, lipopolysacchearide