Li Xiaojun, Jiang Jiaye, Shi Songshan, Bligh S W Annie, Li Yuan, Jiang Yongbo, Huang Dan, Ke Yan, Wang Shunchun
Teaching Experimental Center, Shanghai University of Traditional Chinese Medicine, Shanghai, China; The MOE Key Laboratory for Standardization of Chinese Medicine, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Teaching Experimental Center, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
PLoS One. 2014 Jun 13;9(6):e99697. doi: 10.1371/journal.pone.0099697. eCollection 2014.
Korean mondshood root polysaccharides (KMPS) isolated from the root of Aconitum coreanum (Lévl.) Rapaics have shown anti-inflammatory activity, which is strongly influenced by their chemical structures and chain conformations. However, the mechanisms of the anti-inflammatory effect by these polysaccharides have yet to be elucidated. A RG-II polysaccharide (KMPS-2E, Mw 84.8 kDa) was isolated from KMPS and its chemical structure was characterized by FT-IR and NMR spectroscopy, gas chromatography-mass spectrometry and high-performance liquid chromatography. The backbone of KMPS-2E consisted of units of [→6) -β-D-Galp (1→3)-β-L-Rhap-(1→4)-β-D-GalpA-(1→3)-β-D-Galp-(1→] with the side chain →5)-β-D-Arap (1→3, 5)-β-D-Arap (1→ attached to the backbone through O-4 of (1→3,4)-L-Rhap. T-β-D-Galp is attached to the backbone through O-6 of (1→3,6)-β-D-Galp residues and T-β-D-Ara is connected to the end group of each chain. The anti-inflammatory effects of KMPS-2E and the underlying mechanisms using lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages and carrageenan-induced hind paw edema were investigated. KMPS-2E (50, 100 and 200 µg/mL) inhibits iNOS, TLR4, phospho-NF-κB-p65 expression, phosphor-IKK, phosphor-IκB-α expression as well as the degradation of IκB-α and the gene expression of inflammatory cytokines (TNF-α, IL-1β, iNOS and IL-6) mediated by the NF-κB signal pathways in macrophages. KMPS-2E also inhibited LPS-induced activation of NF-κB as assayed by electrophorectic mobility shift assay (EMSA) in a dose-dependent manner and it reduced NF-κB DNA binding affinity by 62.1% at 200 µg/mL. In rats, KMPS-2E (200 mg/kg) can significantly inhibit carrageenan-induced paw edema as ibuprofen (200 mg/kg) within 3 h after a single oral dose. The results indicate that KMPS-2E is a promising herb-derived drug against acute inflammation.
从朝鲜紫菀（Aconitum coreanum (Lévl.) Rapaics）根部分离得到的朝鲜紫菀根多糖（KMPS）已显示出抗炎活性，其化学结构和链构象对该活性有强烈影响。然而，这些多糖的抗炎作用机制尚未阐明。从KMPS中分离出一种RG-II多糖（KMPS-2E，分子量84.8 kDa），并通过傅里叶变换红外光谱（FT-IR）、核磁共振光谱（NMR）、气相色谱-质谱联用（GC-MS）和高效液相色谱对其化学结构进行了表征。KMPS-2E的主链由[→6)-β-D-吡喃半乳糖(1→3)-β-L-鼠李糖-(1→4)-β-D-吡喃半乳糖醛酸-(1→3)-β-D-吡喃半乳糖-(1→]单元组成，侧链→5)-β-D-阿拉伯糖(1→3, 5)-β-D-阿拉伯糖(1→通过(1→3,4)-L-鼠李糖的O-4连接到主链上。T-β-D-吡喃半乳糖通过(1→3,6)-β-D-吡喃半乳糖残基的O-6连接到主链上，T-β-D-阿拉伯糖连接到每条链的末端基团。使用脂多糖（LPS）刺激的RAW 264.7巨噬细胞和角叉菜胶诱导的后爪水肿模型，研究了KMPS-2E的抗炎作用及其潜在机制。KMPS-2E（50, 100和200 µg/mL）可抑制巨噬细胞中诱导型一氧化氮合酶（iNOS）、Toll样受体4（TLR4）、磷酸化核因子κB p65（phospho-NF-κB-p65）的表达、磷酸化IκB激酶（phosphor-IKK）、磷酸化IκB-α的表达以及IκB-α的降解，以及由NF-κB信号通路介导的炎性细胞因子（肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、iNOS和白细胞介素-6（IL-6））的基因表达。通过电泳迁移率变动分析（EMSA）检测发现，KMPS-2E还以剂量依赖性方式抑制LPS诱导的NF-κB激活，在200 µg/mL时，它使NF-κB与DNA的结合亲和力降低了62.1%。在大鼠中，单次口服给药后3小时内，KMPS-2E（200 mg/kg）可像布洛芬（200 mg/kg）一样显著抑制角叉菜胶诱导的爪水肿。结果表明，KMPS-2E是一种有前景的抗急性炎症的草药衍生药物。
