Anhui University of Chinese Medicine, Hefei, 230012, China; Key Laboratory of Xin'an Medicine, Ministry of Education, Anhui Province Key Laboratory of R&D of Chinese Medicine, Hefei, 230012, China; Anhui Province Key Laboratory of Chinese Medicinal Formula, Hefei, 230012, China.
Anhui University of Chinese Medicine, Hefei, 230012, China; Key Laboratory of Xin'an Medicine, Ministry of Education, Anhui Province Key Laboratory of R&D of Chinese Medicine, Hefei, 230012, China; Anhui Province Key Laboratory of Chinese Medicinal Formula, Hefei, 230012, China.
J Ethnopharmacol. 2021 Jun 28;274:114067. doi: 10.1016/j.jep.2021.114067. Epub 2021 Mar 23.
Achyranthes bidentata Blume (AB) is a traditional Chinese medicine (TCM) widely used as a dietary supplement and anti-arthritis drug. Pharmacological studies have shown that Achyranthes bidentata Blume saponins (ABS) are the main bioactive ingredient. However, the metabolic profile and mechanisms of action of ABS against rheumatic arthritis (RA) remain to be established.
Our main objective was to investigate the metabolic profile and pharmacological activities of ABS against RA.
In this study, an analytical method based on ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) coupled with a metabolism platform was developed for metabolic profiling of ABS in rat liver microsomes and plasma. Then, the in vivo metabolites of ABS and their targets associated with RA were used to construct the network pharmacological analysis. Gene ontology (GO) enrichment, KEGG signaling pathway analyses and pathway network analyses were performed. The therapeutic effect of ABS on RA was further evaluated using an adjuvant arthritis (AA) model and network pharmacology results validated via Western blot.
Overall, 26 and 21 metabolites of ABS were tentatively characterized in rat liver microsomes and plasma, respectively. The metabolic pathways of ABS mainly included M+O, M+O-H, M+O, and M+O-H. Data form network pharmacology analysis suggested that MAPK, apoptosis, PI3K-AKT and p53 signaling pathways contribute significantly to the therapeutic effects of ABS on RA. In pharmacodynamics experiments, ABS ameliorated the symptoms in AA rats in a dose-dependent manner and restored the homeostasis of pro/anti-inflammatory factors. Western blot results further demonstrated a significant ABS-induced decrease in phosphorylation of ERK in the MAPK pathway (P < 0.01).
Application of an analytical method based on UPLC-QTOF/MS, network pharmacology and validation experiments offers novel insights into the components and mechanisms of ABS that contribute to its therapeutic effects against RA, providing useful directions for further research.
牛膝（Achyranthes bidentata Blume）是一种传统的中药，被广泛用作膳食补充剂和抗关节炎药物。药理学研究表明，牛膝皂苷（ABS）是其主要的生物活性成分。然而，ABS 对风湿性关节炎（RA）的代谢谱和作用机制仍有待确定。
本研究旨在探讨 ABS 对 RA 的代谢谱和药理活性。
在这项研究中，我们建立了一种基于超高效液相色谱-四极杆飞行时间质谱（UPLC-QTOF/MS）与代谢物平台相结合的分析方法，用于 ABS 在大鼠肝微粒体和血浆中的代谢谱分析。然后，我们利用 ABS 的体内代谢物及其与 RA 相关的靶标构建网络药理学分析。进行了基因本体（GO）富集、KEGG 信号通路分析和通路网络分析。进一步采用佐剂性关节炎（AA）模型评价 ABS 的治疗效果，并通过 Western blot 验证网络药理学结果。
共在大鼠肝微粒体和血浆中分别推测出 26 种和 21 种 ABS 的代谢产物。ABS 的代谢途径主要包括 M+O、M+O-H、M+O 和 M+O-H。网络药理学分析的数据表明，MAPK、凋亡、PI3K-AKT 和 p53 信号通路对 ABS 治疗 RA 的疗效有重要贡献。在药效学实验中，ABS 以剂量依赖的方式改善 AA 大鼠的症状，并恢复促炎/抗炎因子的平衡。Western blot 结果进一步表明，ABS 可显著降低 MAPK 通路中 ERK 的磷酸化（P<0.01）。
应用基于 UPLC-QTOF/MS、网络药理学和验证实验的分析方法，为 ABS 治疗 RA 的成分和机制提供了新的见解，为进一步研究提供了有用的方向。
