Background Colorectal cancer (CRC) remains a significant clinical challenge, with progression largely driven by tumor microenvironment (TME) remodeling. Cancer-associated fibroblasts (CAFs), key stromal components, promote CRC progression and metastasis via paracrine signaling. Solanum nigrum L. (SNL), a traditional medicinal plant rich in steroidal glycoalkaloids, is recognized for its health-promoting potential. Purpose This study aimed to systematically investigate the bioactive chemical constituents of an aqueous SNL extract, evaluate its biological effects in multiple CRC models, and link the extract's chemical profile to its functional outcomes, focusing on elucidating its anti-metastatic molecular mechanisms. Methods We evaluated the in vivo efficacy of SNL in subcutaneous xenograft, APCMin/+, and experimental metastasis models. Phytochemical analysis identified its major active components. By integrating single-cell RNA sequencing (scRNA-seq) of human CRC with TMT-based proteomics of CRC cells, we identified and validated key molecular targets of SNL. We employed drug-target interaction assays, genetic modulation, and molecular analyses to elucidate its mechanism of action. Results SNL markedly inhibited tumor growth and lung/liver metastasis in vivo. Phytochemical analysis identified solasonine and solamargine as the major glycoalkaloids responsible for the biological activity. Multi-omics analysis identified Annexin A2 (ANXA2) as a key SNL-regulated target. Mechanistically, SNL disrupted CAF-induced signaling, reversed epithelial-mesenchymal transition (EMT), and suppressed invasion by modulating the STAT3-ANXA2-Cortactin axis. While SNL did not bind ANXA2 directly, its active components, solasonine and solamargine, directly interacted with STAT3, blocking its phosphorylation and nuclear translocation, thus repressing ANXA2 transcription. Reduced ANXA2 levels prevented the ANXA2-Cortactin complex formation and inhibited Src/Cortactin phosphorylation, thereby impairing cytoskeletal remodeling and cell motility. Genetic modulation confirmed ANXA2 as a functional mediator of SNL's anti-metastatic effects. Conclusion These findings provide molecular evidence supporting SNL as a source of multifunctional bioactives capable of reshaping the tumor microenvironment and highlight its potential as a plant-derived therapeutic candidate for metastatic colorectal cancer.

Solanum nigrum L. is a widely distributed ethnomedicinal plant with significant antitumor potential, primarily attributed to its abundant steroidal saponins and steroidal glycoalkaloids. However, compositional variability and safety concerns have historically hindered its clinical translation. In this study, we developed a standardized fraction of S. nigrum berries specifically enriched with steroidal saponins and glycoalkaloids (S-SNB), and systematically evaluated its chemical composition, therapeutic potential against non-small cell lung cancer (NSCLC), underly mechanisms, and safety profile. Compared with the conventional aqueous extract, S-SNB exhibited markedly enhanced tumor suppression in A549 and PC-9 xenograft models at substantially lower crude herb equivalent doses. Phytochemical characterization and bioactivity-guided analyses revealed solasonine, solamargine, macrostemonoside B, and solanigroside J as core bioactive constituents, with the latter reported here for their anti-NSCLC activity for the first time. From a mechanistic perspective, proteome-wide target profiling via limited proteolysis–mass spectrometry (LiP-MS) identified glycolytic pathway as a primary node of S-SNB, which was validated by functional metabolic assays showing suppressed glycolytic flux both in vitro and in vivo. Integrated biophysical, biochemical, and computational analyses further converged on PKM2 as a central target of the major active constituents, revealing a non-classical mode of regulation involving coordinated multi-component interactions rather than direct catalytic inhibition. Collectively, these findings demonstrate that enrichment of steroidal saponins and glycoalkaloids enhances both pharmacological potency and mechanistic clarity of S. nigrum against NSCLC, and establishes S-SNB as a chemically defined, safety-validated, and mechanism-informed botanical preparation, providing a robust scientific foundation for its development as an adjunctive oncology therapy.

Solasonine (SS) has been shown to inhibit the proliferation of various malignant tumors, though its effects on lipid metabolism in tumor cells are less understood. This study investigated SS's anti-tumor mechanism in oral squamous cell carcinoma (OSCC) using lipidomics, cell, and animal models. SS inhibited the growth of CAL27 and WSU-HN30 cells and reduced tumor volume in mice. Lipidomic analysis revealed an increase in diglyceride (DG) and a decrease in triglyceride (TG) levels, alongside a reduction in diacylglycerol acyltransferase 1 (DGAT1), key to TG synthesis. SS also induced reactive oxygen species (ROS) production and mitochondrial damage. Molecular docking confirmed SS's interaction with DGAT1, suggesting it prevents DG to TG conversion, inhibiting OSCC proliferation.

Ethnopharmacological relevance: Zuo Gui Wan (ZGW) is a well-known traditional Chinese medicine decoction used for approximately 400 years to treat age-related degenerative conditions, including cognitive impairment in older adults, osteoporosis, and general aging. However, the mechanism of action for ZGW remains unclear. Aims of the study: This study aims to investigate the efficacy of ZGW in improving cognitive function in Alzheimer's disease (AD) animal models and to explore the underlying mechanisms, presenting a novel perspective in the field. Materials and methods: Six-month-old male APP/PS1 mice were divided into three groups that received either metformin (200 mg/kg daily) or ZGW (6 and 12 g/kg daily). High-performance liquid chromatography was conducted for ZGW's quality control. Cognitive function was assessed using the Morris water maze test. Neuronal loss, synaptic plasticity, and beta-amyloid (A beta) deposition were evaluated through Western blot or immunofluorescence staining. The underlying molecular mechanisms were investigated using ELISA, Western blot, qRT-PCR, co-immunoprecipitation assay, ATP assay, and cytochrome c oxidase assay. Results: ZGW, administered in both low and high doses, significantly enhanced cognitive performance, notably decreased neuronal loss and A beta deposition, and reduced levels of A beta 1-40/42. It also inhibited excessive mitochondrial division primarily by suppressing phosphorylated dynamin-related protein 1 (Drp1), especially at high doses of ZGW. Co-immunoprecipitation experiments further confirmed that ZGW inhibited the interaction between A beta and p-Drp1. Furthermore, similar to the effects of the AMP-activated Protein Kinase (AMPK) activator metformin, ZGW led to a marked increase in the mitochondrial DNA copy number and upregulated the AMPK/ PGC-1 alpha/NRF1/TFAM pathway. Improvements in mitochondrial function were evident from the increased ATP production, elevated expression of superoxide dismutase 2, and upregulated cytochrome c oxidase activity. Additionally, the excess byproduct of reactive oxygen species, 4-hydroxy-2-nonenal, decreased in the group treated with ZGW. Conclusion: This study provides compelling evidence that ZGW improves cognitive impairment in APP/PS1 mice by activating AMPK/PGC-1 alpha-regulated mitochondrial bioenergetics and inhibiting A beta-induced mitochondrial fragmentation, highlighting its potential as an effective therapeutic strategy for AD.

Fourteen novel 14,15-diseco-18-nor-pregnane-type steroidal glycosides, mooreanoside A-C (1-3) and mooreanoside F-P (6-16) and two new 14,15-diseco-pregnane-type steroidal glycosides, mooreanoside D-E (4-5) were isolated from the roots of Cynanchum mooreanum Hemsl. Their structures were determined on the basis of spectroscopic and chemical analysis. Compounds 1-6, 8-10, 12-14 and 16 were tested for their immunological activities in vitro against Con-A induced proliferation of mice splenocytes.