粪肠球菌脂磷壁酸对人牙周膜成纤维细胞表达Toll样受体2及白细胞介素-1β的影响

目的探讨体外培养的人牙周膜成纤维细胞（HPDLCs）在粪肠球菌脂磷壁酸（LTA）刺激下表达Toll样受体2（TLR2）及白细胞介素-1β（IL-1β）的情况。方法体外分离培养HPDLCs,采用流式细胞术检测0.1、1、10 μg·mL-1粪肠球菌LTA刺激24 h后HPDLCs表面TLR2表达的改变,酶联免疫吸附法检测12、24、48 h后IL-1β的分泌情况,并用相同质量浓度的大肠杆菌内毒素作为对照;用TLR2中和抗体预处理HPDLCs 1 h,观察1 μg·mL-1 LTA刺激24 h后其IL-1β的分泌量。结果LTA可致HPDLCs表面TLR2表达增加（P<0.05）;与对照组相比,粪肠球菌LTA可致HPDLCs分 泌IL-1β增加（P<0.05）,刺激12 h后可检测到IL-1β,48 h内呈上升趋势。TLR2中和抗体对粪肠球菌LTA诱导HPDLCs 分泌IL-1β无明显封闭作用。结论粪肠球菌LTA可引起HPDLCs表面TLR2表达及IL-1β分泌增加。

Study on the expression of Toll like receptor 2 and interleukin-1 beta induced by Enterococcus faecalis lipoteichoic acid on human periodontal ligament cells

Objective To investigate the expression of Toll like receptor 2(TLR2) and interleukin-1β(IL-1β) of cultured human periodontal ligament cells(HPDLCs) activated by Enterococcus faecalis(E.faecalis) lipoteichoic acid(LTA).Methods HPDLCs that were obtained from the periodontal tissues of healthy humans were maintained in pro-per condition.Flow cytometry was used to detect the expression of TLR2 on normal HPDLCs and infectious HPDLCs which were incubated with 0.1,1,10 μg·mL-1 E.faecalis LTA for 24 h.IL-1β was detected by enzyme linked immunosorbent assay(ELISA) after incubating with LTA of the above concentration for 12,24 and 48 h or pretreated with TLR2 neutralizing antibody for 1 h and then co-cultured with 1 μg·mL-1 LTA for 24 h.Results E.faecalis LTA pro-moted the expression of TLR2 in normal HPDLCs.The difference had statistical significance(P<0.05).IL-1β secretion could be detected 12 h after stimulation with LTA and increasingly escalate within 48 h(P<0.05).TLR2 neutralizing antibody had no evident effect on IL-1β generation stimulating by E.faecalis LTA.Conclusion E.faecalis LTA can increase the expression of TLR2 and IL-1β in normal HPDLCs.