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作 者:漆晓玲[1,2] 赵蕾[3] 陈珊珊[1] 孟姝[3] 吴亚菲[3]
机构地区:[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
出 处:《华西口腔医学杂志》2016年第2期194-199,共6页West China Journal of Stomatology
基 金:国家自然科学基金(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·m L-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m RNA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·m L-1组的RANTES m RNA和1 h、200 ng·m L-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m RNA表达量及分形素m RNA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m RNA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m RNA表达量及分形素的m RNA表达量均降低,24 h时,仅Pg-LPS质量浓度为500 ng·m L-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·m L-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。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 PgLPS(200, 500, and 1 000 ng·m L-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 m RNA levels of RANTES and fractalkine. Results The RANTES protein levels and m RNA levels, as well as fractalkine m RNA 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 m RNA in 200 ng·m L-1 group of 12 h and RANTES protein in 200 ng·m L-1 group of 1 h. The expression levels of RANTES m RNA and fractalkine m RNA were highest in 1 000 ng·m L-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, m RNA, and fractalkine m RNA decreased 6 h after stimulation, and were significantly higher than those in the control group(P〈0.05) in the 500 ng·m L-1 group of 24 h. There was a significant difference between the expression of fractalkine m RNA in 1 000 ng·m L-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.
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