不同fimA基因型牙龈卟啉单胞菌对人脐静脉内皮细胞分泌相关蛋白及细胞因子的影响

Effects of different fimA genotypes of Porphyromonas gingivalis on the secretion related proteins and cytokines in human umbilical vein endothelial cells

背景:牙龈卟啉单胞菌感染是近年发现与动脉粥样硬化等多种疾病发生有关的危险因素,但不同fimA基因型的牙龈卟啉单胞菌致病能力不同,积极探明牙龈卟啉单胞菌高危菌株对于完善动脉粥样硬化诊疗机制具有重要作用。目的:探讨不同fimA基因型牙龈卟啉单胞菌对人脐静脉内皮细胞分泌单核细胞趋化蛋白1、白细胞介素6、内皮素1/一氧化氮的影响。方法:厌氧条件下对Ⅳ、Ⅱ、ⅠfimA基因型牙龈卟啉单胞菌进行培养,应用感染复数值为100感染人脐静脉内皮细胞,同时设阳性对照组为牙龈卟啉单胞菌脂多糖刺激人脐静脉内皮细胞、阴性对照组人脐静脉内皮细胞仅加细胞培养液培养。采用酶联免疫吸附法检测各组培养2,6,24 h后人脐静脉内皮细胞培养上清液中单核细胞趋化蛋白1、白细胞介素6、内皮素1/一氧化氮水平。结果与结论:除阴性对照组外,其他4组人脐静脉内皮细胞培养上清液中白细胞介素6、内皮素1/一氧化氮、单核细胞趋化蛋白1水平逐渐增高,且ⅣfimA基因型刺激组白细胞介素6、内皮素1/一氧化氮水平高于Ⅱ、ⅠfimA基因型刺激组及阳性对照组、阴性对照组(P<0.05);ⅡfimA基因型刺激组单核细胞趋化蛋白1水平高于Ⅳ、ⅠfimA基因型刺激组、阴性对照组(P<0.05)。结果表明,不同fimA基因型牙龈卟啉单胞菌刺激人脐静脉内皮细胞功能紊乱机制存在一定差异,Ⅱ、ⅣfimA基因上调单核细胞趋化蛋白1、白细胞介素6、内皮素1/一氧化氮分泌能力较强,可能在引起血管内皮细胞功能紊乱中占主导地位。

BACKGROUND:Porphyromonas gingivalis infection is a risk factor that has been found to be related to atherosclerosis and other diseases in recent years.However,different fimA genotypes of Porphyromonas gingivalis have different pathogenic abilities.Actively explore Porphyromonas gingivalis high-risk bacterial strains play an important role in improving the diagnosis and treatment mechanism of atherosclerosis.OBJECTIVE:To investigate the effects of different fimA genotypes of Porphyromonas gingivalis on the secretion of monocyte chemoattractant protein-1,interleukin-6,and endothelin-1/nitric oxide in human umbilical vein endothelial cells.METHODS:Porphyromonas gingivalis genotypes Ⅳ,Ⅱ,and Ⅰ fimA were cultured under anaerobic conditions,and the multiplicity of infection was 100 to infect human umbilical vein endothelial cells.In the positive control group,Porphyromonas gingivalis lipopolysaccharide was used to stimulate human umbilical vein endothelial cells,and in the negative control group,human umbilical vein endothelial cells were cultured with cell culture medium only.Enzyme-linked immunosorbent assay was used to detect the levels of monocyte chemoattractant protein 1,interleukin 6,endothelin 1/nitric oxide in the culture supernatant of human umbilical vein endothelial cells after 2,6,and 24 hours in each group.RESULTS AND CONCLUSION:Except for the negative control group,the levels of interleukin 6,endothelin 1/nitric oxide,and monocyte chemoattractant protein 1 in the culture supernatant of human umbilical vein endothelial cells in the other four groups gradually increased.The levels of interleukin 6,endothelin 1/nitric oxide in the Ⅳ fimA genotype stimulation group were higher than those in the Ⅱ and Ⅰ fimA genotype stimulation groups and the positive control group and the negative control group(P<0.05).The level of monocyte chemoattractant protein 1 in the Ⅱ fimA genotype stimulation group was higher than that in the Ⅳ,Ⅰ fimA genotype stimulation group and the negative control group(P<0.05).The results indicate that there are certain differences in the dysfunction mechanisms of Porphyromonas gingivalis with different fimA genotypes in stimulating human umbilical vein endothelial cells.The Ⅱ fimA genotype and Ⅳ fimA gene have a strong ability to up-regulate the secretion of monocyte chemoattractant protein 1,interleukin 6,and endothelin 1/nitric oxide,and may play a dominant role in causing vascular vein endothelial cell dysfunction.