RESUMO
One of the traditional prescriptions for treating lung diseases, Jiegeng decoction (JGT), is still unknown in terms of its chemical makeup and mechanism. In this study, Q-Exactive-Orbitrap MS technology was used to identify the chemical constituents of JGT, and metabolomics was used to examine the effect of JGT on metabolites in the lung tissue of mice with acute lung injury (ALI) model. The potential biomarkers were screened by fold change (FC) > 1.5 or FC < 0.67 and P < 0.05, and enriched for metabolic pathways. A total of 40 compounds, including triterpenoid saponins, flavonoids and glycosides, were identified by mass spectrometry analysis of JGT. All animal experiments were approved by the Experimental Animal Ethics Committee of Tianjin University of Traditional Chinese Medicine (No. TCM-LAEC2021106). The results showed that JGT improved the lung coefficient, and lung tissue morphology of mice with ALI, lowered the levels of malondialdehyde (MDA), tumor necrosis factor α (TNF-α), and interleukin 6 (IL-6) in bronchoalveolar lavage fluid (BALF), and reduced myeloperoxidase (MPO) content in lung tissue. The metabolomic results showed that JGT could regulate 22 metabolites associated with ALI, among which leukotriene D4, docosapentaenoic acid, hypoxanthine, L-5-oxoproline, and other metabolites were mainly associated with the body′s inflammatory response and oxidative stress, and were enriched in the pathways of glutathione metabolism, purine metabolism, and primary bile acid biosynthesis. This study analyzed the potential mechanism of JGT in the treatment of ALI through metabolomics, providing an important theoretical basis for the clinical application of JGT.
RESUMO
BACKGROUND: The florets of Carthamus tinctorius L. (safflower) serve as the source of a reputable herbal medicine targeting gynecological diseases. Conventional investigations regarding the quality control of safflower, however, mainly focused on the secondary metabolites with primary metabolites ignored. PURPOSE: To holistically evaluate the quality difference of safflower samples collected from five different producing regions by multiple chemical and biological approaches with both the primary and secondary metabolites considered. METHODS: A precursor ions list-triggered data-dependent MS2 approach was established by ultra-high performance liquid chromatography/Q-Orbitrap mass spectrometry (UHPLC/Q-Orbitrap MS) to comprehensively identify the secondary metabolites from safflower. Primary metabolites were identified by various 1D and 2D nuclear magnetic resonance (NMR) experiments. Similarity evaluation and quantitative assays of all the characterized primary metabolites and a quinochalcone C-glycoside (QCG) marker, hydroxysafflor yellow A (HSYA), were performed by quantitative 1H NMR (qNMR) using an external standard method. Multiple in vitro models with respect to the antioxidant, anti-platelet aggregation, and antioxidant stress injury effects, were assayed to determine the efficacy differences. RESULTS: Totally thirteen primary metabolites (including one nucleoside, two sugars, five organic alkali/acids, and five amino acids) and 135 secondary metabolites (97 QCGs and 38 flavonoids) could be identified or tentatively characterized from safflower. Good chemical consistency was observed between the commercial safflower samples and a standard safflower sample, with similarity varying in the range of 0.95â0.99. The results from qNMR-oriented quantitative experiments (thirteen primary metabolites and HSYA) and biological assays indicated the quality of safflower samples from Xinjiang (XJ-2 and XJ-4), Hunan (HuN-1 and HuN-2), and Sichuan (SC), was comparable to the standard safflower sample. CONCLUSION: The integration of multiple chemical (using two analytical platforms, UHPLC/Q-Orbitrap MS and NMR) and biological (four in vitro models) approaches by determining both the primary and secondary metabolites demonstrated a powerful strategy that could facilitate the holistic quality evaluation of traditional Chinese medicine.