Fingerprinting of Sophora alopecuroides Extract and Its Anti-inflammatory Action Spectrum-effect Relationship
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目的:建立苦豆子总黄酮、总生物碱、总多糖配伍后的指纹图谱,研究不同配伍的化学成分与细胞活力以及抗炎之间的关联性,明确不同配伍对细胞活力的影响以及抗炎作用的物质基础.方法:采用高效液相色谱(HPLC)建立苦豆子总黄酮、总生物碱、总多糖配伍后的指纹图谱,以 LPS 刺激 RAW 264.7建立炎症细胞模型,对不同配伍组合对炎症细胞模型肿瘤坏死因子(TNF-α)水平以及以细胞活力进行检测,采用正交偏最小二乘判别分析(OPLS-DA)研究苦豆子化学成分与抗炎作用的谱效关系,筛选苦豆子抗炎作用的物质基础.结果:苦豆子生物碱类成分HPLC 指纹图谱共有峰有8个,指认出5个共有峰,分别是槐果碱、槐定碱、苦参碱、氧化槐果碱和氧化苦参碱;黄酮类成分HPLC 指纹图谱共有峰有15个,指认出3个共有峰,分别为槲皮素、紫铆因和芹菜素.相关性分析结果表明,槐果碱、槐定碱、氧化槐果碱和氧化苦参碱与脂多糖(LPS)诱导的小鼠单核巨噬细胞(RAW 264.7)细胞释放的炎症因子TNF-α 水平呈显著负相关,槲皮素与细胞活力呈负相关,紫铆因、芹菜素与细胞活力呈正相关,槐定碱、氧化苦参碱与细胞活力呈负相关,苦参碱、槐果碱、氧化槐果碱与细胞活力呈正相关.结论:槐果碱、槐定碱、氧化槐果碱和氧化苦参碱可能是苦豆子抗炎的物质基础,可为挖掘苦豆子的质量标志物以及药材质量控制提供参考.
Objective:To establish the fingerprint of total flavonoids,total alkaloids and total polysac-charides of Sophora alopecuroides to study the association between different compatibility chemical components and cell viability and anti-inflammatory,and to clarify the effect of different compatibility on cell viability and the material basis of anti-inflammatory effect.Methods:High performance liquid chromatography(HPLC)was used to establish the fingerprint of total flavonoids,total alkaloids and total polysaccharides of S.alopecuroides.The inflammatory cell model of RAW264.7 was stimulated by LPS.The levels of tumor necrosis factor(TNF-α)and cell vitality of the inflammatory cell model were detected by different combinations.Grey orthogonal partial least squares discriminant analysis(OPLS-DA)were used to study the spectral relationship between chemical components and anti-in-flammatory effects of S.alopecuroides,and to screen the material basis of anti-inflammatory effects of S.alopecuroides.Results:There were 8 peaks in the HPLC fingerprint of alkaloids of S.alopecuroi-des,and 5 peaks were identified,which were sophocarpine,sophoridine,matrine,oxysophocarpine and oxymatrine.There were 15 peaks in the HPLC fingerprint of flavonoids,and 3 peaks were identi-fied,namely quercetin,butein and apigenin.The results of correlation analysis showed that sophocar-pine,sophoridine,oxysophocarpine and oxymatrine were significantly negatively correlated with the levels of inflammatory cytokine TNF-α released by RAW 264.7 cells induced by LPS.Quercetin was negatively correlated with cell viability,butein and apigenin were positively correlated with cell viabili-ty,sophodinine and oxymatrine were negatively correlated with cell viability,while matrine,sopho-carpine and oxysophocarpine oxide were positively correlated with cell viability.Conclusion:Ophocar-pine,sophoridine,oxysophocarpine and oxymatrine may be the material basis for the anti-inflammato-ry of S.alopecuroides,which can provide a reference for the mining of quality markers of S.alope-curoides and the quality control of medicinal materials.
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