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  

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

作　　者：于鑫 王月秋[2] 李明恒 苏勤[4] 许海平[4] 邢路[4] 

机构地区：[1]大连市口腔医院牙体牙髓科,大连116021 [2]江苏省口腔医院第四门诊部,南京210029 [3]大连市口腔医院口腔颌面外科,大连116021 [4]口腔疾病研究国家重点实验室,四川大学,成都610041

出　　处：《华西口腔医学杂志》2012年第3期325-328,共4页West China Journal of Stomatology

基　　金：四川省科技攻关基金资助项目(2006Z09-038)

摘　　要：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、1μg.mL-1、10μg.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1μg.mL-1LPS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,24h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1μg.mL-1LPS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。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（P0.05）.In the three groups,the expression of RANKL was significantly different（P0.05）.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.

关 键 词：TOLL样受体 人牙周膜成纤维细胞 脂多糖 细胞核因子-κB受体活化因子配基 

分 类 号：R781.34[医药卫生—口腔医学]