Isorhamnetin inhibits Prevotella intermedia lipopolysaccharide-induced production of interleukin-6 in murine macrophages via anti-inflammatory heme oxygenase-1 induction and inhibition of nuclear factor-κB and signal transducer and activator of transcription 1 activation
Isorhamnetin inhibits Prevotella intermedia lipopolysaccharide-induced production of interleukin-6 in murine macrophages via anti-inflammatory heme oxygenase-1 induction and inhibition of nuclear factor-κB and signal transducer and activator of transcription 1 activation
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DOI：
10.1111/jre.12054
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发表时间：
2013-12-01
影响因子：
3.5
通讯作者：
Kim, S. J.
中科院分区：
文献类型：
--
作者：
Jin, J. Y.;Choi, E. Y.;Kim, S. J.
背景与意义白细胞介素-6（IL-6）是一种重要的促炎细胞因子，在牙周病的发病机制中起重要作用。因此，针对抑制IL-6的宿主调节剂似乎在减弱牙周病进展和潜在地改善疾病易感性方面是有益的。在目前的研究中，我们研究了类黄酮异鼠李素对IL-6在小鼠巨噬细胞的生产刺激与脂多糖（LPS）从中间普雷沃氏菌，一种病原体牵连在炎症性牙周病，其mechanisms of action.Material and MethodsLipopolysaccharide从中间普雷沃氏菌ATCC 25611分离使用标准的热酚-水方法。收集培养物上清液并测定IL-6。我们使用实时PCR定量IL-6和血红素加氧酶-1（HO-1）mRNA的表达。采用免疫印迹法检测HO-1蛋白的表达和信号蛋白的水平。用ELISA法检测细胞核因子-B（NF-B）的DNA结合活性。结果异鼠李素能显著下调中间对虾LPS诱导的IL-6的产生及其mRNA在RAW 264. 7细胞中的表达。异鼠李素在基因转录和翻译水平上上调中间对虾LPS刺激细胞HO-1的表达。此外，锡原卟啉IX抑制HO-1活性可阻断异鼠李素对IL-6产生的抑制作用。异鼠李素不能阻止LPS激活c-Jun N-末端激酶或p38通路。异鼠李素在抑制B-降解的水平上不抑制NF-B的转录活性。异鼠李素通过抑制NF-B p50亚基的核转位和DNA结合活性抑制NF-B信号传导，减弱信号转导子和转录激活子1信号传导。我们认为异鼠李素可能有助于阻断由IL-2介导的宿主破坏过程。6，并可能是一个非常有效的调节宿主反应，在治疗炎症性牙周病。需要在牙周炎动物模型中进行进一步研究以更好地评估异鼠李素作为治疗牙周病的新型药物的潜力。
Background and ObjectiveInterleukin-6 (IL-6) is a key proinflammatory cytokine that has been considered to be important in the pathogenesis of periodontal disease. Therefore, host-modulatory agents directed at inhibiting IL-6 appear to be beneficial in terms of attenuating periodontal disease progression and potentially improving disease susceptibility. In the current study, we investigated the effect of the flavonoid isorhamnetin on the production of IL-6 in murine macrophages stimulated with lipopolysaccharide (LPS) from Prevotella intermedia, a pathogen implicated in inflammatory periodontal disease, and its mechanisms of action.Material and MethodsLipopolysaccharide from P.intermedia ATCC 25611 was isolated using the standard hot phenol-water method. Culture supernatants were collected and assayed for IL-6. We used real-time PCR to quantify IL-6 and heme oxygenase-1 (HO-1) mRNA expression. The expression of HO-1 protein and the levels of signaling proteins were monitored using immunoblot analyses. The DNA-binding activity of nuclear factor-B (NF-B) was analyzed using ELISA-based assay kits.ResultsIsorhamnetin significantly down-regulated P.intermedia LPS-induced production of IL-6 as well as its mRNA expression in RAW264.7 cells. Isorhamnetin up-regulated the expression of HO-1 at both gene transcription and translation levels in cells stimulated with P.intermedia LPS. In addition, inhibition of HO-1 activity by tin protoporphyrin IX blocked the inhibitory effect of isorhamnetin on IL-6 production. Isorhamnetin failed to prevent LPS from activating either c-Jun N-terminal kinase or p38 pathways. Isorhamnetin did not inhibit NF-B transcriptional activity at the level of inhibitory B- degradation. Isorhamnetin suppressed NF-B signaling through inhibition of nuclear translocation and DNA binding activity of NF-B p50 subunit and attenuated signal transducer and activator of transcription 1 signaling.ConclusionAlthough further research is required to clarify the detailed mechanism of action, we propose that isorhamnetin may contribute to blockade of the host-destructive processes mediated by IL-6 and could be a highly efficient modulator of the host response in the treatment of inflammatory periodontal disease. Further research in animal models of periodontitis is required to better evaluate, the potential of isorhamnetin as a novel agent for treating periodontal disease.