背景:野菊花(FCI),菊花的花,是一种在中国流行的治疗炎症性疾病的中药(TCM)。FCI也是一种功能性食品,并被广泛用作清热解毒的凉茶。
目的:基于最佳采收期,探索FCI的质量控制标记。
方法:首先,基于UPLC-Q-TOF/MS的非靶向代谢组学应用于探索在芽阶段(BS)收集的FCI的化学概况,初始阶段(IS),中国八个耕地的盛开阶段(FS)和最终阶段(ES)。随后,脂多糖(LPS)诱导的RAW264.7细胞炎症模型和角叉菜胶诱导的大鼠爪水肿模型用于证实在IS/FS收集的FCI的抗炎作用。然后,采用UPLC-PDA靶向代谢组学方法对开花期9种具有抗炎活性的成分(7种黄酮类化合物和2种酚酸)进行定量分析(VIP>4)。最后,根据不同开花期FCI中9种活性成分的变化,将ROC曲线结合PCA分析用于筛选FCI的质量标记。
结果:IS/FS处的FCI具有几乎相同的化学特性,但与BS和ES的完全不同。在开花发育过程中,FCI中共有32种成分发生了变化,包括类黄酮和酚酸。与ES相比,IS/FS下大多数不同成分的含量最高或更高,表明FCI的最佳收获期应为IS/FS。FCI提取物可以有效抑制LPS诱导的RAW264.7细胞中一氧化氮(NO)的产生,并调节角叉菜胶诱导的爪水肿模型大鼠细胞因子和PGE2的异常水平。定量分析结果表明,在开花发育过程中,FCI中酚酸和黄酮的变化趋势不同,但在从八个栽培区收集的所有FCI中,大多数在IS/FS的含量高于ES的含量,除了TC,安徽。最后,linarin,木犀草素,根据其AUC值在ROC中的贡献和PCA分析的聚类,选择芹菜素和3,5-二咖啡酰基奎尼酸作为Q标记。
结论:我们的研究证明了FCI的最佳收获期,并根据生长进程对活性成分的影响,使用非靶向/靶向代谢组学确定了FCI的多成分Q标记。研究结果不仅大大提高了FCI资源的利用率,提高了FCI产品的质量控制,而且还提供了新的策略来识别FCI的Q标记。
BACKGROUND: Flos Chrysanthemi Indici (FCI), the flower of Chrysanthemum Indicum L., is a popular traditional Chinese medicine (TCM) for treatment of inflammatory diseases in
China. FCI is also a functional food, and is widely used as herbal tea for clearing heat and detoxicating.
OBJECTIVE: To explore quality control markers of FCI based on the optimal harvest period.
METHODS: First, UPLC-Q-TOF/MS based untargeted metabolomics was applied to explore the chemical profiles of FCIs collected at bud stages (BS), initial stages (IS), full bloom stages (FS) and eventual stages (ES) from eight cultivated regions in
China. Subsequently, lipopolysaccharide (LPS)-induced RAW264.7 cell inflammatory model and carrageenan-induced rat paw
edema model were used to confirm the anti-inflammatory effect of FCIs collected at IS/FS. Then, UPLC-PDA targeted metabolomics was used to quantitatively analyze 9 constituents with anti-inflammatory activity (7 flavonoids and 2 phenolic acids) changed significantly (VIP > 4) during flowering stages. Finally, ROC curves combined with PCA analysis based on the variation of 9 active constituents in FCIs from different flowering stages were applied to screen the quality markers of FCI.
RESULTS: FCIs at IS/FS had almost same chemical characteristics, but quite different from those at BS and ES. A total of 32 constituents in FCIs including flavonoids and phenolic acids were changed during flowering development. Most of the varied constituents had the highest or higher contents at IS/FS compared with those at ES, indicating that the optimal harvest period of FCI should be at IS/FS. FCI extract could effectively suppress nitric oxide (NO) production in LPS-induced RAW264.7 cells and regulate the abnormal levels of cytokines and PGE2 in carrageenan-induced paw
edema model rat. The results of quantitatively analysis revealed that the variation trends of phenolic acids and flavonoids in FCIs were different during flowering development, but most of them had higher contents at IS/FS than those at ES in all FCIs collected from eight cultivated regions, except one sample from Anhui. Finally, linarin, luteolin, apigenin and 3,5-dicaffeoylquinic acid were selected as the Q-markers based on the contribution of their AUC values in ROC and clustering of PCA analysis.
CONCLUSIONS: Our study demonstrates the optimal harvest period of FCI and specifies the multi-constituents Q-markers of FCI based on the influence of growth progression on the active constituents using untargeted/targeted metabolomics. The findings not only greatly increase the utilization rate of FCI resources and improve quality control of FCI products, but also offer new strategy to identify the Q-markers of FCI.