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

doi:10.3969/j.issn.1000-1182.2012.03.025

华西口腔医学杂志

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

于鑫1 王月秋2 李明恒3 苏勤4 许海平4 邢路4

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

收稿日期:2012-06-25 修回日期:2012-06-25 出版日期:2012-06-01 发布日期:2012-06-01
通讯作者: 苏勤，Tel：028-85501439

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 Yu Xin1, Wang Yueqiu2, Li Mingheng3, Su Qin4, Xu Haiping4, Xing Lu4.

Yu Xin1, Wang Yueqiu2, Li Mingheng3, Su Qin4, Xu Haiping4, Xing Lu4

1. Dept. of Endodontics, Dalian Stomatological Hospital, Dalian 116021,China; 2. The Fourth Outpatient Department, Stomatological Hospital of Jiangsu Province, Nanjing 210029, China;3. Dept. of Oral and Maxillofacial Surgery, Dalian Stomatological Hospital, Dalian 116021, China; 4. State Key Laboratory
of Oral Diseases, Sichuan University, Chengdu 610041, China

Received:2012-06-25 Revised:2012-06-25 Online:2012-06-01 Published:2012-06-01
Contact: Su Qin，Tel：028-85501439

摘要/Abstract

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、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。

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

Abstract:

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-κB ligand（RANKL） expression of human periodontal 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 antibody 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 approximately 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 expression in HPDLFs induced by LPS. Anti-TLR4 antibody has better inhibition effect to RANKL expression of HPDLFs stimulated by LPS than anti-TLR2.

Key words: Toll-like receptors, human periodontal ligament fibroblasts, lipopolysaccharide, receptor activator of nuclear factor-κB ligand

于鑫王月秋李明恒苏勤许海平邢路 . 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[J]. 华西口腔医学杂志, doi: 10.3969/j.issn.1000-1182.2012.03.025.

Yu Xin1, Wang Yueqiu2, Li Mingheng3, Su Qin4, Xu Haiping4, Xing Lu4. 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 Yu Xin1, Wang Yueqiu2, Li Mingheng3, Su Qin4, Xu Haiping4, Xing Lu4.[J]. West China Journal of Stomatology, doi: 10.3969/j.issn.1000-1182.2012.03.025.

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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 Yu Xin1, Wang Yueqiu2, Li Mingheng3, Su Qin4, Xu Haiping4, Xing Lu4.

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