Toll样受体2和4在脂多糖诱导人牙周膜成纤维细胞表达细胞核因子-κB受体活化因子配基中的作用

目的 观察在脂多糖（LPS）刺激下,人牙周膜成纤维细胞（HPDLFs）中Toll样受体2（TLR2）和Toll样受体4（TLR4）表达水平的抑制对其表达细胞核因子-κB受体活化因子配基（RANKL）的影响。方法 选用100 ng·mL-1、1 μg·mL-1、10 μg·mL-1大肠杆菌LPS分别刺激HPDLFs,刺激6、12、24、48 h后,采用酶联免疫吸附试验（ELISA）检测HPDLFs表达RANKL的水平。分别运用不同滴度的anti-TLR2+anti-TLR4、anti-TLR2、anti-TLR4抗体预处理HPDLFs,观察
1 μg·mL-1 LPS刺激下,其RANKL表达水平的变化。结果 LPS刺激HPDLFs 6 h后,即可检测到RANKL的表达,24 h达到顶峰,然后逐渐下降;各LPS质量浓度组的规律基本一致。分别用anti-TLR2+anti-TLR4、anti-TLR2、anti-TLR4抗体预处理HPDLFs,在1 μg·mL-1 LPS刺激下,其产生RANKL的水平明显下降（P<0.05）;3组中,RANKL的表达水平有明显差异（P<0.05）,其中anti-TLR2+anti-TLR4抗体处理组RANKL表达量最少,anti-TLR4抗体处理组次之,anti-TLR2
抗体处理组RANKL的表达量最高。结论TLR2、TLR4均参与了LPS诱导HPDLFs表达RANKL的过程;与anti-TLR2抗体相比,anti-TLR4抗体能更有效地抑制LPS刺激后HPDLFs表达RANKL的能力。

Effect of Toll-like receptor 2 and Toll-like receptor 4 on expression of receptor activator of nuclear factor-kappaB ligand in human periodontal ligament fibroblasts induced by lipopolysaccharide

Objective The aim of this study was to survey the influence of Toll-like receptor 2(TLR2) and Tolllike receptor 4(TLR4) repression to receptor activator of nuclear factor-B ligand(RANKL) expression of human perio-dontal ligament fibroblasts(HPDLFs) under the stimulation of lipopolysaccharide(LPS).Methods The level of RANKL in HPDLFs stimulated by 100 ng·mL-1,1 μg·mL-1 and 10 μg·mL-1 Escherichia coli(E.coli) LPS after 6,12,24 and 48 h was detected by enzyme linked immunosorbent assay(ELISA).The level of RANKL in HPDLFs stimulated by 1 μg·mL-1 E.coli LPS after pretreatment with different titre anti-TLR2+anti-TLR4,anti-TLR2 and anti-TLR4 anti-body were observed respectively.Results RANKL was detected at 6 h after stimulation with LPS,and the levels of these cytokine were highest at 24 h,and then gradually decreased.The regularity of each LPS concentration was appro-ximately similar.After pretreatment with anti-TLR2+anti-TLR4,anti-TLR2 and anti-TLR4 antibody,the level of RANKL was significantly decreased under the stimulation of 1 μg·mL-1 LPS(P<0.05).In the three groups,the expression of RANKL was significantly different(P<0.05).The level of RANKL in anti-TLR2+anti-TLR4 antibody pretreatment group was the lowest,the level in anti-TLR4 antibody pretreatment group was higher,and the level in anti-TLR2 antibody pretreatment group was the highest.Conclusion TLR2 and TLR4 participate in the process of RANKL expres-sion in HPDLFs induced by LPS.Anti-TLR4 antibody has better inhibition effect to RANKL expression of HPDLFs stimulated by LPS than anti-TLR2.