Discovery and verification of anti-inflammatory-related quality markers in the aerial part of Bupleurum scorzonerifolium by UPLC-Q-TOF-MS/MS and in RAW 264.7 cells and a zebrafish model

Phytochem Anal. 2024 Mar;35(2):336-349. doi: 10.1002/pca.3291. Epub 2023 Oct 3.

Abstract

Introduction: The root of Bupleurum scorzonerifolium Willd. (BS) is officially recognized in the Chinese Pharmacopoeia. In contrast, the aerial part of BS (ABS), accounting for 80% of BS, is typically discarded, causing potential waste of medicinal resources. ABS has shown benefits in the treatment of inflammation-related diseases in China and Spain, and the material basis underlying its anti-inflammatory effects must be systematically elucidated for the rational use of ABS.

Objective: We aimed to screen and validate the anti-inflammatory quality markers (Q-markers) of ABS and to confirm the ideal time for ABS harvesting.

Methods: The chemical components and anti-inflammatory effects of ABS from 10 extracted parts were analyzed by UPLC-Q-TOF-MS/MS and in a lipopolysaccharide (LPS)-induced cell model. Anti-inflammatory substances were screened by Pearson bivariate analysis and gray correlation analysis, and the anti-inflammatory effects were verified in a zebrafish tail-cutting inflammation model. HPLC was applied to measure the Q-marker contents of ABS in different harvesting periods.

Results: Ten ABS extracts effectively alleviated the increase in LPS-induced proinflammatory cytokines in RAW 264.7 cells. Forty components were identified from them, among which 27 were common components. Eight components were correlated with anti-inflammatory effects, which were confirmed to reverse the expression of proinflammatory and anti-inflammatory factors in a zebrafish model. Chlorogenic acid, hypericin, rutin, quercetin, and isorhamnetin can be detected by HPLC, and the maximum contents of these five Q-markers were obtained in the sample harvested in August.

Conclusion: The anti-inflammatory Q-markers of ABS were elucidated by chromatographic-pharmacodynamic-stoichiometric analysis, which served as a crucial basis for ABS quality control.

Keywords: Bupleurum scorzonerifolium; UPLC-Q-TOF-MS/MS; aerial part; anti-inflammatory; quality marker.

MeSH terms

  • Animals
  • Anti-Inflammatory Agents / analysis
  • Anti-Inflammatory Agents / pharmacology
  • Bupleurum* / chemistry
  • Chromatography, High Pressure Liquid
  • Inflammation / drug therapy
  • Lipopolysaccharides / pharmacology
  • Mice
  • Plant Components, Aerial / chemistry
  • RAW 264.7 Cells
  • Tandem Mass Spectrometry*
  • Zebrafish

Substances

  • Lipopolysaccharides
  • Anti-Inflammatory Agents