Lipopolysaccharide of Aggregatibacter actinomycetemcomitans up-regulates inflammatory cytokines, prostaglandin E2 synthesis and osteoclast formation in interleukin-1 receptor antagonist-deficient mice
Lipopolysaccharide of Aggregatibacter actinomycetemcomitans up-regulates inflammatory cytokines, prostaglandin E2 synthesis and osteoclast formation in interleukin-1 receptor antagonist-deficient mice
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DOI：
10.1111/jre.12065
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发表时间：
2013-12-01
影响因子：
3.5
通讯作者：
Noguchi, T.
中科院分区：
文献类型：
--
作者：
Mizutani, H.;Ishihara, Y.;Noguchi, T.
背景与目的白细胞介素(IL)-1受体拮抗剂（Ra）与IL-1受体结合，抑制IL-1活性。然而，IL-1Ra是否在牙周病中起保护作用尚不清楚。本研究的目的是比较IL-1Ra敲除（KO）和野生型（WT）小鼠对牙周细菌脂多糖（LPS）的促炎细胞因子产生、破骨细胞形成和骨吸收的反应。材料与方法取13周龄IL-1Ra KO和WT小鼠腹腔巨噬细胞（Ms）。腹腔Ms分别加入或不加入10g/mL放线菌聚合杆菌LPS培养24h。采用酶联免疫吸附法（ELISA）测定骨膜上清液（PM-SF）中IL-1 α (IL-1)、IL-1 β (IL-1)、肿瘤坏死因子- (TNF-)和IL-6的水平。取小鼠骨髓细胞，用PM-SF刺激9d，然后用TRAP染色。基于融合指数计算trap阳性的多核巨细胞形成的频率。采用显微计算机断层扫描分析pm - sf刺激的颅骨骨吸收情况，并用苏木精、伊红和TRAP染色对颅骨进行组织学分析。实时荧光定量PCR检测骨髓细胞环氧化酶-2 （Cox2）、前列腺素受体EP4 （EP4）及Rank mrna的表达，ELISA检测前列腺素E2 （PGE(2)）的产生。结果放线菌多糖刺激IL-1Ra KO小鼠PM-SF后，IL-1、IL-1、TNF-和IL-6水平显著升高约4- (p
Background and ObjectiveThe interleukin (IL)-1 receptor antagonist (Ra) binds to IL-1 receptors and inhibits IL-1 activity. However, it is unclear whether the IL-1Ra plays a protective role in periodontal disease. The purpose of this study was to compare IL-1Ra knockout (KO) and wild-type (WT) mice in regard to proinflammatory cytokine production, osteoclast formation and bone resorption in response to periodontal bacterial lipopolysaccharide (LPS).Material and MethodsPeritoneal macrophages (Ms) were obtained from 13-wk-old IL-1Ra KO and WT mice. Peritoneal Ms were cultured with or without 10g/mL of Aggregatibacter actinomycetemcomitans LPS for 24h. The levels of IL-1alpha (IL-1), IL-1beta (IL-1), tumor necrosis factor- (TNF-) and IL-6 were measured in periotoneal M phi s supernatant fluid (PM-SF) using an ELISA. Bone marrow cells were obtained from the mice and stimulated with PM-SF for 9d, then stained with TRAP. The frequency of TRAP-positive multinucleated giant cell formation was calculated based on a fusion index. PM-SF-stimulated calvarial bone resorption was analyzed using micro-computed tomography, and calvarial histological analysis was performed using hematoxylin and eosin and TRAP staining. The expression of cyclooxygenase-2 (Cox2), prostanoid receptor EP4 (Ep4) and Rank mRNAs in bone marrow cells were measured using real-time quantitative PCR, while prostaglandin E2 (PGE(2)) production was determined by ELISA.ResultsThe levels of IL-1, IL-1, TNF- and IL-6 in IL-1Ra KO mice PM-SF stimulated with A.actinomycetemcomitans LPS were significantly increased by approximately 4- (p