高糖对牙龈卟啉单胞菌LPS引发牙周膜细胞炎症反应的影响

Impact of High Glucose on Porphyromonas Gingivalis LPS induced Inflammation of Periodontal Ligament Cells

目的:观察高糖对牙龈卟啉单胞菌(Porphyromonas gingivalis,P.gingivalis)重要毒力因子脂多糖(lipopolysaccharide,LPS)引发宿主牙周膜细胞释放炎症细胞因子和对核因子-κB(Nuclear factor-κB,NF-κB)表达、活化的影响。方法:人牙周膜细胞(human periodontal ligament cells,hPDLCs)培养于葡萄糖生理浓度中作为对照组,P.gingivalis LPS(1μg/mL)、高糖(25 mmol/L)、高糖+P.gingivalis LPS刺激细胞6 h和12 h后,酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测上清液中白细胞介素-1β(Interleukin-1β,IL-1β)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的释放,实时荧光定量PCR(quantitative real time PCR,qRT-PCR)和Western-blot检测NF-κB p65和磷酸化NF-κB p65(phospho-NF-κB p65,p-NF-κB p65)在hPDLCs中的表达水平。结果:高糖和P.gingivalis LPS可分别促进hPDLCs表达IL-lβ和TNF-α、促进NF-κB p65表达和活化,高糖和P.gingivalis LPS的共刺激可以增强这一促进作用。结论:高糖可能通过对NF-κB的活化增强P.gingivalis LPS致hPDLCs的炎症作用。

Objective:To investigate the impact of high glucose on inflammatory mediators release and NF-κB expression and on activation of human periodontal ligament cells induced by P.gingivalis lipopolysaccharide.Methods:Primary cultured hPDLCs were challenged in P.gingivalis LPS(1μg/mL),high glucose(25mmol/L),and high glucose+P.gingivalis LPS for 6 and 12 hours.The hPDLCs were cultured in physical concentration of glucose as control.The expression of TNF-αand IL-1βwere detected by ELISA.The expression of NF-κB p65 and p-NF-κB p65 were measured by qRT-PCR and western-blot.Results:High glucose and P.gingivalis LPS could promote the expression of inflammatory cytokines TNF-αand IL-1β,and promote the expression and activation of NF-κB p65 in hPDLCs.The co-stimulation of high glucose and P.gingivalis LPS could enhance the expression of inflammatory cytokines and activation of NF-κB p65.Conclusion:High glucose may enhance the inflammation induced by P.gingivalis LPS through activation of NF-κB.