Traditional Chinese Medicine Processing Technology Innovation Center of Hebei Province, School of Pharmacy, Hebei University of Chinese Medicine, Shijiazhuang 050200, China; International Joint Research Center on Resource Utilization and Quality Evaluation of Traditional Chinese Medicine of Hebei Province, Hebei University of Chinese Medicine, Shijiazhuang 050200, China.
Traditional Chinese Medicine Processing Technology Innovation Center of Hebei Province, School of Pharmacy, Hebei University of Chinese Medicine, Shijiazhuang 050200, China; Hebei Chemical and Pharmaceutical College, Shijiazhuang 050026, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2022 Dec 15;1213:123531. doi: 10.1016/j.jchromb.2022.123531. Epub 2022 Nov 8.
The rhizome of Dioscorea nipponica Makino (RDN) is a widely used herbal medicine, which has significant anti-inflammatory activities on various inflammatory diseases. However, the bioactive compositions responsible for the anti-inflammatory activity of RDN are still unknown. This study aimed to identify the anti-inflammatory bioactive compounds in RDN using high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-Q/TOF-MS), quantitative analysis of multiple components by single marker (QAMS) and chemometric methods. Firstly, an HPLC-Q/TOF-MS method was employed for identification of bioactive steroidal saponins in RND, and a total of twelve steroid saponins were identified. Then, QAMS method was employed to determine the contents of seven bioactive steroidal saponins, including protodioscin, protogracillin, methyl protodioscin, pseudoprotodioscin, pseudoprogracillin, dioscin and gracillin in RND samples using dioscin as the reference analyte. The anti-inflammatory effects of RDN samples were then evaluated by inhibition of NO production in LPS-induced RAW264.7 cells. Furthermore, chemometric methods, including Pearson correlation analysis and partial least squares regression (PLSR) were employed to investigate the correlations between chemical components and anti-inflammatory activities, and explore the potential anti-inflammatory bioactive compounds of RDN. The results indicated that protodioscin, dioscin and gracillin were selected as the major anti-inflammatory compounds in RND. The further verification experiments showed that protodioscin, dioscin and gracillin exhibited great inhibition on NO production with IC values (the half maximal inhibitory concentration) of 0.712 μM, 0.469 μM and 0.815 μM, respectively. They also significantly reduced the levels of TNF-α, IL-1β, and IL-6 in LPS-induced RAW264.7 cells. The present study provided evidences for the anti-inflammatory activity of RND and identification of the anti-inflammatory components in RDN.
怀地黄的根茎是一种广泛使用的草药，对各种炎症性疾病具有显著的抗炎活性。然而，负责怀地黄抗炎活性的生物活性成分仍不清楚。本研究旨在使用高效液相色谱-四极杆飞行时间质谱（HPLC-Q/TOF-MS）、多成分单标志物定量分析（QAMS）和化学计量学方法，从怀地黄中鉴定出具有抗炎活性的生物活性化合物。首先，采用 HPLC-Q/TOF-MS 方法鉴定怀地黄中的生物活性甾体皂苷，共鉴定出 12 种甾体皂苷。然后，采用 QAMS 方法测定怀地黄样品中 7 种生物活性甾体皂苷（原薯蓣皂苷、原纤细薯蓣皂苷、甲基原薯蓣皂苷、伪原薯蓣皂苷、伪纤细薯蓣皂苷、薯蓣皂苷和美丽薯蓣皂苷）的含量，以薯蓣皂苷为参照分析物。然后，通过抑制 LPS 诱导的 RAW264.7 细胞中 NO 的产生来评价怀地黄样品的抗炎作用。此外，采用 Pearson 相关分析和偏最小二乘回归（PLSR）等化学计量学方法，研究化学组分与抗炎活性之间的相关性，探讨怀地黄的潜在抗炎生物活性成分。结果表明，原薯蓣皂苷、薯蓣皂苷和美丽薯蓣皂苷被选为怀地黄中的主要抗炎化合物。进一步的验证实验表明，原薯蓣皂苷、薯蓣皂苷和美丽薯蓣皂苷对 NO 产生的抑制作用较强，IC 值（半最大抑制浓度）分别为 0.712 μM、0.469 μM 和 0.815 μM，它们还显著降低了 LPS 诱导的 RAW264.7 细胞中 TNF-α、IL-1β 和 IL-6 的水平。本研究为怀地黄的抗炎活性提供了证据，并鉴定了怀地黄中的抗炎成分。
