Porphyromonas gingivalis Induces Receptor Activator of NF-κB Ligand Expression in Osteoblasts through the Activator Protein 1 Pathway
Porphyromonas gingivalis Induces Receptor Activator of NF-κB Ligand Expression in Osteoblasts through the Activator Protein 1 Pathway
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DOI：
10.1128/iai.72.3.1706-1714.2004
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发表时间：
2004-03
影响因子：
3.1
通讯作者：
N. Okahashi;H. Inaba;I. Nakagawa;Taihei Yamamura;M. Kuboniwa;K. Nakayama;S. Hamada;A. Amano
中科院分区：
文献类型：
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作者：
N. Okahashi;H. Inaba;I. Nakagawa;Taihei Yamamura;M. Kuboniwa;K. Nakayama;S. Hamada;A. Amano
牙龈卟啉单胞菌是一种重要的牙周病原菌，与炎症性牙槽骨吸收密切相关，据报道，该菌的几种成分如脂多糖可刺激细胞因子的产生，促进炎症性骨破坏。我们研究了活牙龈卟啉单胞菌感染对成骨细胞产生细胞因子的影响。RT-PCR和Real-time PCR分析显示牙龈卟啉单胞菌感染诱导小鼠原代成骨细胞核因子κB受体激活因子（NF-κB）配体（RANKL）mRNA表达。白细胞介素-6的产生也受到刺激，但骨保护素没有。p38丝裂原活化蛋白激酶抑制剂SB 20350、经典丝裂原活化蛋白激酶激酶（MEK 1/2）抑制剂PD 98059、磷脂酰肌醇3激酶抑制剂渥曼青霉素和NF-κB抑制剂羧苄青霉素-亮氨酰-亮氨酰-亮氨酸不能阻止牙龈卟啉单胞菌诱导的RANKL表达。未检测到NF-κB-alpha抑制剂的降解;然而，姜黄素（一种激活蛋白1（AP-1）抑制剂）可阻止牙龈卟啉单胞菌感染诱导的RANKL产生。蛋白质印迹分析显示，磷酸化的c-Jun，AP-1的一个组成部分，发生在感染的细胞，和c-Fos结合的寡核苷酸含有AP-1的共识网站的分析也证明AP-1激活感染的成骨细胞。感染牙龈卟啉单胞菌KDP 136（精氨酸和赖氨酸特异性半胱氨酸蛋白酶的同基因缺陷突变体）不会刺激RANKL的产生。这些结果表明，牙龈卟啉单胞菌感染通过AP-1信号通路诱导成骨细胞中RANKL表达，生物体的半胱氨酸蛋白酶参与RANKL的产生。
ABSTRACT Porphyromonas gingivalis, an important periodontal pathogen, is closely associated with inflammatory alveolar bone resorption, and several components of the organism such as lipopolysaccharides have been reported to stimulate production of cytokines that promote inflammatory bone destruction. We investigated the effect of infection with viable P. gingivalis on cytokine production by osteoblasts. Reverse transcription-PCR and real-time PCR analyses revealed that infection with P. gingivalis induced receptor activator of nuclear factor κB (NF-κB) ligand (RANKL) mRNA expression in mouse primary osteoblasts. Production of interleukin-6 was also stimulated; however, osteoprotegerin was not. SB20350 (an inhibitor of p38 mitogen-activated protein kinase), PD98059 (an inhibitor of classic mitogen-activated protein kinase kinase, MEK1/2), wortmannin (an inhibitor of phosphatidylinositol 3 kinase), and carbobenzoxyl-leucinyl-leucinyl-leucinal (an inhibitor of NF-κB) did not prevent the RANKL expression induced by P. gingivalis. Degradation of inhibitor of NF-κB-alpha was not detectable; however, curcumin, an inhibitor of activator protein 1 (AP-1), prevented the RANKL production induced by P. gingivalis infection. Western blot analysis revealed that phosphorylation of c-Jun, a component of AP-1, occurred in the infected cells, and an analysis of c-Fos binding to an oligonucleotide containing an AP-1 consensus site also demonstrated AP-1 activation in infected osteoblasts. Infection with P. gingivalis KDP136, an isogenic deficient mutant of arginine- and lysine-specific cysteine proteinases, did not stimulate RANKL production. These results suggest that P. gingivalis infection induces RANKL expression in osteoblasts through AP-1 signaling pathways and cysteine proteases of the organism are involved in RANKL production.