Anti-inflammatory effect of gallic acid-conjugated chitooligosaccharides in lipopolysaccharide-stimulated RAW 264.7 macrophages

Van-Hoai Bui, Hong-Tham N Vo, Thanh-Tuan Duong, Se-Kwon Kim, Dai-Nghiep Ngo
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Abstract

The aim of the present study is to investigate the anti-inflammatory effect of gallic acid grafted onto COS chains (GA-COS), focusing on the reduction of nitric oxide production, downregulation of inflammatory signals such as inducible nitric oxide synthase (iNOS), gene expression of cytokines such as TNF-α, IL-1β, IL-6, and nuclear factor kappa B (NF-κB) signalling, including the p50 and p65 subunits. The anti-inflammatory effect is mediated through the reduction of nitric oxide production and downregulation of inflammatory proteins such as inducible nitric oxide synthase (iNOS), gene expression of cytokines like TNF-α, IL-1β, IL-6, and nuclear factor kappa B (NF-κB) signalling, including the p50 and p65 subunits. Target proteins were identified by western blot analysis with specific monoclonal antibodies. The levels of gene expression were determined by the RT-PCR method. The results demonstrate that GA-COS effectively reduces nitric oxide generation and downregulates iNOS protein and cytokine expression and NF-κB signalling in lipopolysaccharide (LPS) -induced RAW 264.7 cells. GA-COS exhibits significantly enhanced anti-inflammatory activity compared to the free chitooligosaccharide chain. This study lays the groundwork for future research to demonstrate that GA-COS holds significant potential as a novel compound for the prevention of inflammatory diseases.
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没食子酸结合壳寡糖在脂多糖刺激的 RAW 264.7 巨噬细胞中的抗炎作用
本研究旨在探讨接枝到 COS 链(GA-COS)上的没食子酸的抗炎作用,重点是减少一氧化氮的产生、下调炎症信号(如诱导型一氧化氮合酶(iNOS))、细胞因子(如 TNF-α、IL-1β、IL-6)的基因表达以及核因子卡巴 B(NF-κB)信号(包括 p50 和 p65 亚基)。抗炎作用是通过减少一氧化氮的产生和下调炎症蛋白(如诱导型一氧化氮合酶(iNOS))、细胞因子(如 TNF-α、IL-1β、IL-6)的基因表达以及核因子卡巴B(NF-κB)信号传导(包括 p50 和 p65 亚基)来实现的。使用特异性单克隆抗体进行蛋白印迹分析,确定目标蛋白。基因表达水平是通过 RT-PCR 方法测定的。结果表明,在脂多糖(LPS)诱导的 RAW 264.7 细胞中,GA-COS 能有效减少一氧化氮的生成,下调 iNOS 蛋白和细胞因子的表达以及 NF-κB 信号的传递。与游离壳寡糖链相比,GA-COS 的抗炎活性明显增强。这项研究为今后的研究奠定了基础,以证明 GA-COS 作为一种新型化合物在预防炎症性疾病方面具有巨大潜力。
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