牙龈卟啉单胞菌对人脐静脉内皮细胞分泌IL-1β和IL-6的影响研究

摘要

AIM: To observe the effects of Porphyromonas gingivalis on expression of IL-6 and IL-1β inrnhuman umbilical vein endothelial cells (HLVECs). METHODS: Primary clutures of HUVECs were derived by enzyme digestion method. Passage 5 HUVECs were stimulated by P. Gingivalis supernatant for 6, 12, 18 and 24 h. At specific time points, mRNA was extracted and subjected for RT-PCR to detect IL-6 and IL-1β mRNA expression. HUVEC culture supernatant was collected and subjected for ELISA to measure IL-6 and IL-1β protein levels. RESULTS: P. Gingivalis supernatant stimulation significantly increased IL-1β mRNA expression at 12 and 18 h (P<0.05) , and it also significantly increased IL-6 mRNA at 12 h (P<0.05). Furthermore, P. Gingivalis stimulation resulted in significantly higher IL-1β secretion at 18 and 24 h (P < 0.05 ) . And IL-6 protein level was significantly augmented at 12, 18 and 24 h ( P < 0. 05). CONCLUSION: P. Gingivalis can stimulate the expression of IL-6 and IL-1β in human umbilical vein endothelial cells.%目的:研究牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)上清液对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)分泌IL-1β、IL-6的影响.方法:酶消化法原代培养HUVECs,并对细胞进行来源鉴定,用5×106CFU/mL的Pg上清液刺激HUVECs 6、12、18、24 h后,RT-PCR检测IL-1β、IL-6 mRNA的表达;同时ELISA方法测定IL-1β及IL-6蛋白的水平.结果:Pg上清液刺激HUVECs后,IL-1 βmRNA水平在12、18h时蛋白明显高于对照组(P＜0.05);IL-6 mRNA水平在12 h的表达明显高于对照组(P＜0.05).IL-1β蛋白在18、24 h的表达明显高于对照组(P ＜0.05);IL-6蛋白在12、18、24h3个时点均明显高于对照组(P＜0.05).结论:5×106 CFU/mL Pg上清液可促进人脐静脉内皮细胞IL-1β和IL-6的基因及蛋白表达,提示Pg感染可能在As的发生发展中起一定作用.