A total of 31 compounds were isolated from the ethyl acetate and n-butanol fractions of Anchusa italica Retz., which contained one ursane triterpenoid, 2α,3β,19α-trihydroxy-23-formyl-urs-12-en-28,21β-olide (1), and five norisoprenoids: (2R,6R,9S)-9-hydroxy-4-megastigmen-3-one-2-O-β-D-glucopyranoside (3); (2R,6S,9S)-9-hydroxy-megastigman-4,7-dien-3-one-2-O-β-D-glucopyranoside (4); (+)-isololiolide β-D-glucopyranoside (5); (2S,8R)-loliolide β-D-glucopyranoside (6a); and (2R,8S)-loliolide β-D-glucopyranoside (6b). It also contained 25 known compounds (2 and 7–30). The chemical structures of the compounds, inclusive of their absolute configurations, were ascertained using spectroscopic methods such as NMR, HR-MS, and quantum chemical calculations (computational NMR and ECD), in combination with relevant literature data. Moreover, the chemotaxonomic significance of the isolated substances was discussed, with compounds 1, 2, and 7–13 potentially broadening the application of triterpenes as taxonomic markers for the classification of the genus Anchusa.

The root bark of Morus alba L. is commonly used as a natural antioxidant; however, its active constituents and underlying molecular mechanisms remain unclear. In this study, a bioactivity-guided isolation approach was employed to identify antioxidant substances from the root bark of Morus alba L. and to investigate their protective effects against oxidative damage in HaCaT cells. Using techniques such as silica gel column chromatography and semi-preparative HPLC, combined with NMR and HR-ESI-MS analysis, 22 compounds were isolated and identified from the dichloromethane extract of Morus alba L. root bark, including Diels–Alder adducts, flavonoids, and benzofurans. Among them, compounds 1 and 2 are new compounds, while compounds 12 and 16 were isolated from this plant for the first time. Bioactivity screening revealed that Kuwanon A (compound 17) exhibited significant cytoprotective effects in an H₂O₂-induced HaCaT cell injury model, effectively scavenging intracellular reactive oxygen species (ROS), restoring mitochondrial function, and enhancing the activities of antioxidant enzymes such as SOD and GSH. Further studies indicated that H₂O₂ induced ferroptosis in HaCaT cells, characterized by abnormal Fe²⁺ levels, lipid peroxidation, and elevated levels of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α). Kuwanon A significantly ameliorated these pathological changes. Consistently, ELISA and Astral DIA quantitative proteomics analyses demonstrated that Kuwanon A specifically upregulates the expression of the sulfurtransferase NFS1, thereby promoting the expression of the core antioxidant enzyme GPX4 and the iron storage protein ferritin-H, collectively inhibiting ferroptosis. This study elucidates that Kuwanon A is a key active component responsible for the antioxidant and anti-inflammatory effects of Morus alba L. root bark, and its mechanism is closely associated with regulating the NFS1-mediated ferroptosis defense pathway.

Aconitum tanguticum (Maxim.) Stapf is a Tibetan medicinal plant with low toxicity. To obtain the major alkaloid heteratisine (HTS) for bioactivity evaluation, this study established a method combining deep eutectic solvent (DES) with ultrasound-assisted extraction. COSMO-RS screening identified menthol-octanoic acid (1:2) as the optimal solvent, giving a content of 0.321 mg/g, approximately 9.17 times that of ethanol. Extraction optimization was performed via response surface methodology on a 29-run Box–Behnken design, providing a validated optimum of 0.499 mg/g at 256.8 W, 30.5 min, 1:30.3 g/mL, and 39.7 °C. The fitting behavior of three machine learning models and quadratic polynomial regression (QPR) was compared on a 75-run non-structured dataset. XGBoost and GPR achieved high in-sample fitting performance (R² = 0.983 and 0.994), whereas QPR and Random Forest showed limited fitting (0.529 and 0.546). Condition-grouped nested validation demonstrated that none of the models could extrapolate beyond the observed dataset. Thus, the ML analyses were framed as in-sample diagnostic tools, with GPR offering uncertainty estimates to identify data-sparse regions. In HEK293T-TRPV1 cells, HTS pretreatment reduced capsaicin-induced Ca²⁺ signaling. In general, this study provides a DES-based extraction approach and a multi-model comparison framework for the utilization of A. tanguticum.