DataSheet1_Simultaneous quantitative analysis and in vitro anti-arthritic effects of five polyphenols from Terminalia chebula.pdf

Background: The fruit of Terminalia chebula has been widely used for a thousand years for treating diarrhea, ulcers, and arthritic diseases in Asian countries. However, the active components of this Traditional Chinese medicine and their mechanisms remain unclear, necessitating further investigation.
Objectives: To perform simultaneous quantitative analysis of five polyphenols in T. chebula and evaluate their anti-arthritic effects including antioxidant and anti-inflammatory activity in vitro.
Materials and methods: Water, 50% water-ethanol, and pure ethanol were used as extract solvents. Quantitative analysis of gallic acid, corilagin, chebulanin, chebulagic acid, and ellagic acid in the three extracts was performed using high-performance liquid chromatography (HPLC). Antioxidant activity was assessed by the 2,2-diphenylpicrylhydrazyl (DPPH) radical-scavenging assay, and anti-inflammatory activity was evaluated by detecting interleukin (IL)-6 and IL-8 expression in IL-1β-stimulated MH7A cells.
Results: The 50% water-ethanol solvent was the optimal solvent yielding the highest total polyphenol content, and the concentrations of chebulanin and chebulagic acid were much higher than those of gallic acid, corilagin, and ellagic acid in the extracts. The DPPH radical-scavenging assay showed that gallic acid and ellagic acid were the strongest antioxidative components, while the other three components showed comparable antioxidative activity. As for the anti-inflammatory effect, chebulanin and chebulagic acid significantly inhibited IL-6 and IL-8 expression at all three concentrations; corilagin and ellagic acid significantly inhibited IL-6 and IL-8 expression at high concentration; and gallic acid could not inhibit IL-8 expression and showed weak inhibition of IL-6 expression in IL-1β-stimulated MH7A cells. Principal component analysis indicated that chebulanin and chebulagic acid were the main components responsible for the anti-arthritic effects of T. chebula.
Conclusion: Our findings highlight the potential anti-arthritic role of chebulanin and chebulagic acid from T. chebula.

背景：诃子（Terminalia chebula）的果实在亚洲国家已被广泛应用千年，用于治疗腹泻、溃疡及关节炎类疾病。然而，这味中药的活性成分及其作用机制仍未明确，亟待开展进一步研究。
研究目的：同时定量分析诃子中的五种多酚类成分，并体外评价其抗氧化、抗炎等抗关节炎活性。
材料与方法：以去离子水、50%水乙醇溶液及无水乙醇作为提取溶剂。采用高效液相色谱法（HPLC）对三种提取物中的没食子酸（gallic acid）、柯里拉京（corilagin）、诃子宁（chebulanin）、诃子酸（chebulagic acid）及鞣花酸（ellagic acid）进行定量分析。抗氧化活性通过2,2-二苯基-1-苦基苯肼（DPPH）自由基清除实验进行评价；抗炎活性则通过检测白细胞介素-1β（IL-1β）刺激的MH7A细胞中白细胞介素-6（IL-6）与白细胞介素-8（IL-8）的表达水平进行评估。
结果：50%水乙醇溶液为最优提取溶剂，其所获提取物的总多酚含量最高；提取物中诃子宁与诃子酸的浓度显著高于没食子酸、柯里拉京及鞣花酸。DPPH自由基清除实验结果显示，没食子酸与鞣花酸的抗氧化活性最强，其余三种成分的抗氧化活性相当。在抗炎活性层面，诃子宁与诃子酸在三个浓度梯度下均可显著抑制IL-6与IL-8的表达；柯里拉京及鞣花酸仅在高浓度下可显著抑制IL-6与IL-8的表达；而没食子酸对IL-8的表达无抑制作用，对IL-1β刺激的MH7A细胞中IL-6的表达仅表现出微弱的抑制效果。主成分分析结果表明，诃子宁与诃子酸是诃子发挥抗关节炎活性的核心物质基础。
结论：本研究结果证实，诃子来源的诃子宁与诃子酸具有潜在的抗关节炎应用价值。