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

Abstract

IntroductionThe 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.ObjectiveWe aimed to screen and validate the anti‐inflammatory quality markers (Q‐markers) of ABS and to confirm the ideal time for ABS harvesting.MethodsThe 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.ResultsTen 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.ConclusionThe anti‐inflammatory Q‐markers of ABS were elucidated by chromatographic‐pharmacodynamic‐stoichiometric analysis, which served as a crucial basis for ABS quality control.