Prevotella intermedia lipopolysaccharide stimulates release of nitric oxide by inducing expression of inducible nitric oxide synthase
Prevotella intermedia lipopolysaccharide stimulates release of nitric oxide by inducing expression of inducible nitric oxide synthase
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DOI：
10.1111/j.1600-0765.2004.00757.x
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发表时间：
2004-12-01
影响因子：
3.5
通讯作者：
Choi, IS
中科院分区：
文献类型：
--
作者：
Kim, SJ;Ha, MS;Choi, IS
目的：研究炎症性牙周病的主要致病因子中间普氏杆菌脂多糖对小鼠巨噬细胞系RAW264.7产生一氧化氮(NO)和诱导型一氧化氮合酶(INOS)表达的影响。我们还试图阐明中间假单胞菌脂多糖诱导NO产生的信号机制。材料和方法：中间假单胞菌ATCC 25611脂多糖的制备采用标准的热酚-水法。通过测定培养上清液中亚硝酸盐的积累量来检测NO的产生。结果：在没有额外刺激的情况下，中间拟青霉能够诱导iNOS表达，并刺激NO的释放，证实了转录因子核因子-kappaB(NF-kappaB)和微管聚合在NO产生中的重要作用。NO的产生需要L-精氨酸，但不需要蛋白激酶C或蛋白酪氨酸激酶的激活。结论：本研究清楚地表明，在没有其他刺激的情况下，中间假单胞菌能够完全诱导RAW264.7细胞诱导型一氧化氮合酶的表达和一氧化氮的产生。中间假单胞菌的脂多糖促进NO产生的能力可能在炎症性牙周病的发病机制中起重要作用。
Objectives: The purpose of this study was to examine the effects of lipopolysaccharide from Prevotella intermedia, a major cause of inflammatory periodontal disease, on the production of nitric oxide (NO) and expression of inducible nitric oxide synthase (iNOS) in the murine macrophage cell line RAW264.7. We also attempted to throw light on the signaling mechanisms involved in P. intermedia lipopolysaccharide-induced NO production.Material and methods: Lipopolysaccharide from P. intermedia ATCC 25611 was prepared by the standard hot phenol-water method. NO production was assayed by measuring the accumulation of nitrite in culture supernatants. Western blot analysis of iNOS and analysis of reverse transcription-polymerase chain reaction (RT-PCR) products were carried out.Results: We found that P. intermedia lipopolysaccharide can induce iNOS expression and stimulate the release of NO without additional stimuli and demonstrated an important role of the transcription factor nuclear factor-kappaB (NF-kappaB) and microtubule polymerization in NO production. The production of NO required L-arginine but not activation of protein kinase C or protein tyrosine kinase.Conclusions: The present study clearly shows that P. intermedia lipopolysaccharide fully induced iNOS expression and NO production in RAW264.7 cells in the absence of other stimuli. The ability of P. intermedia lipopolysaccharide to promote the production of NO may be important in the pathogenesis of inflammatory periodontal disease.