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姜科植物根茎蛋白对巨噬样 RAW264.7 细胞一氧化氮生成的抑制作用。

Inhibition of nitric oxide production in the macrophage-like RAW 264.7 cell line by protein from the rhizomes of Zingiberaceae plants.

机构信息

Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.

出版信息

Prep Biochem Biotechnol. 2013;43(1):60-78. doi: 10.1080/10826068.2012.697958.

Abstract

Nitric oxide (NO) plays a key role in the pathogenesis of inflammation and has been implicated in endotoxin-induced tissue injury. Zingiberaceae is a family of indigenous plants of tropical regions, many of which have traditionally been used as anti-inflammatory agents. Here, the ability of crude protein extracts from the rhizomes of 15 Zingiberaceae species to inhibit NO production in the RAW 264.7 cell line after co-stimulation with lipopolysaccharide (LPS) and interferon-gamma (IFN-γ) was evaluated. The crude protein extract of Zingiber ottensii Valeton exhibited the highest inhibitory activity, with an IC(50) value of 38.6 ± 0.34 µg protein/mL, and also suppressed the LPS- and rm-interferon (IFN)-γ-mediated increase in the inducible nitric oxide synthase (iNOS), interleukin (IL)-6, and tumor necrosis factor (TNF)-α mRNA transcript expression levels, suggesting the interference was mediated at the transcriptional level. This strong anti-inflammatory activity may have the potential to be developed as a therapeutic compound. Analytical sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry revealed four main protein bands, including a likely lectin, superoxide dismutase, and cysteine protease, in the fractions related to the antioxidant activity.

摘要

一氧化氮(NO)在炎症发病机制中起关键作用,并与内毒素诱导的组织损伤有关。姜科是热带地区土生土长的植物科,其中许多植物传统上被用作抗炎剂。在这里,评估了来自 15 种姜科植物根茎的粗蛋白提取物在脂多糖(LPS)和干扰素-γ(IFN-γ)共刺激后对 RAW 264.7 细胞系中 NO 产生的抑制能力。姜科植物奥滕氏姜(Zingiber ottensii Valeton)的粗蛋白提取物表现出最高的抑制活性,IC(50)值为 38.6±0.34µg 蛋白/mL,并且还抑制了 LPS 和 rm-干扰素(IFN)-γ介导的诱导型一氧化氮合酶(iNOS)、白细胞介素(IL)-6 和肿瘤坏死因子(TNF)-α mRNA 转录表达水平的增加,表明这种干扰是在转录水平上进行的。这种强烈的抗炎活性可能有潜力被开发为一种治疗化合物。分析性十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)和质谱分析显示,在与抗氧化活性相关的馏分中存在四种主要的蛋白质带,包括一种可能的凝集素、超氧化物歧化酶和半胱氨酸蛋白酶。

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