牙龈卟啉单胞菌脂多糖对人脐静脉内皮细胞表达小凹蛋白-1的影响

Effect of Porphyromonas Gingivalis Lipopolysaccharide on Expression of Cav-1 in HUVECs

目的:初步研究牙龈卟啉单胞菌脂多糖(P.gingivalis lipopolysaccharide,P.g-LPS)对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)小凹蛋白-1(caveolin-1,Cav-1)表达的影响,并探讨P.g-LPS与动脉粥样硬化(atherolerosis,AS)之间的关系。方法:体外培养HUVECs,选择不同浓度的P.g-LPS(0.5、1.0、5.0、10.0 mg/L)诱导刺激6、18、24、36 h,采用5-乙炔基-2’脱氧尿嘧啶核苷(5-ethynyl-2’-deoxyuridine,EdU)法检测P.g-LPS对HUVECs增殖的影响,酶联免疫吸附实验(enzyme linked immunosorbent assay,ELISA)检测HUVECs表达Cav-1的量,采用逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)检测Cav-1 mRNA的表达。结果:P.g-LPS与HUVECs共培养后,增殖活力较对照组下降,并随着P.g-LPS浓度增加,HUVECs增殖活力明显下降(P<0.05)。与对照组比较,P.g-LPS诱导HUVECs Cav-1蛋白表达在18 h可升高,24、36 h持续高表达,36 h表达水平最高(P<0.05);P.g-LPS诱导HUVECs Cav-1 mRNA表达在18 h可升高,24、36 h随着浓度的增加,Cav-1 mRNA表达量增加(P<0.001)。结论:P.g-LPS能够明显抑制HUVECs的增殖活性,上调细胞Cav-1的蛋白以及分子水平表达。提示P.g-LPS感染可能引起内皮细胞功能障碍,从而加速AS的发生与发展。

Objective:To study the effect of P.gingivalis lipopolysaccharide(P.g-LPS)on the expression of caveolin-1(Cav-1)in human umbilical vein endothelial cells(HUVECs),and to explore the relationship between P.g-LPS and atherosclerosis(AS).Methods:HUVECs were cultured in vitro,and different concentrations of P.g-LPS(0.5,1.0,5.0,10.0 mg/L)were selected to induce for 6,18,24 and 36 hours.The effects of P.g-LPS on the proliferation of HUVECs were detected by EdU method,the expression of Cav-1 in HUVECs was detected by ELISA,and the expression of Cav-1 was detected by RT-PCR.Results:After co-culture of P.g-LPS and HUVECs,the proliferation activity of HUVECs decreased compared with the control group.With the increase of P.g-LPS concentration,the proliferation activity of HUVECs decreased significantly(P<0.05).Compared with the control group,the expression of Cav-1 protein in HUVECs induced by P.g-LPS increased for 18 hours,sustained high expression for 24 and 36 hours,with the highest expression level at 36 hours(P<0.05);the expression of Cav-1 gene in HUVECs induced by P.g-LPS increased for 18 hours,and the expression of Cav-1 gene increased with the increase of concentration at 24 and 36 hours(P<0.001).Conclusion:P.g-LPS can significantly inhibit the proliferation activity of HUVECs and up-regulate the protein and molecular level of Cav-1 cells.It is suggested that P.g-LPS infection may cause endothelial cell disfunction,thereby accelerating the occurrence and development of AS.