牙龈卟啉单胞菌对脐静脉血管内皮细胞表达IL-8和MCP-1 mRNA的影响

Effect of Porphyromonas gingivalis on the mRNA expression of IL-8 and MCP-1 in human umbilical vein endothelial cells

目的 :在转录水平上观察牙龈卟啉单胞菌对脐静脉血管内皮细胞 (humanumbilicalveinen dothelialcells,HUVECs)表达白细胞介素 - 8(IL - 8)和单核细胞趋化蛋白 - 1(MCP - 1)的影响。方法 :厌氧培养Pg ,原代培养HUVECs,RNA抽提 ,逆转录 -聚合酶链式反应 (RT -PCR)和mRNA比色定量法检测IL - 8和MCP - 1基因表达。结果 :HUVECs基础表达IL - 8和MCP - 1mRNA ,Pg以剂量依赖的方式增强IL - 8和MCP - 1mRNA的表达 ,Pg感染后 1hIL - 8mRNA开始增高 ,3h达到高峰 ,并持续到 5h。而MCP - 1的mRNA从Pg感染后 1h开始增高 ,3h达到高峰 ,5h出现下降趋势。结论 :Pg在转录水平上增强HUVECs表达IL - 8和MCP - 1,这种上调作用可能是牙周病早期基因水平调控炎症细胞募集的重要因素 ,可能是牙周疾病的炎症反应和免疫反应中主要调控机制之一。

AIM: To study the effect of Porphyromonas gingivalis ( Pg ) on interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) expression at transcription level in human umbilical vein endothelial cells (HUVECs). METHODS:Anaeropack method was employed for Pg . Primary culture of HUVEC was taken. The extraction of total RNA, reverse transcription-polymerase chain reaction (RT-PCR) and mRNA colorimetric quantification were used for mRNA of IL-8 and MCP-1 . RESULTS:HUVEC expressed IL-8 and MCP-1 mRNA constructively. Pg dose-dependently increased the mRNA amount of IL-8 and MCP-1 in ECHUV. The up-regulation of IL-8 mRNA started at 1h after Pg infection, reached maximal level at 3h, and kept at this level at 5h; whereas elevated MCP-1 mRNA was found at 1h, peaked at 3h, and then decreased at 5h. CONCLUSION: Pg increased the expression of IL-8 and MCP-1 in HUVEC at transcription level. The up-regulated effect of IL-8 and MCP-1 mRNA in the endothelial cells could be the key mechanism in controlling the recruitment of leukocytes in periodontitis, and it might play an important role in the inflammatory and immunological reactions of periodontal disease.