Cinobufagin

This study aims to investigate the therapeutic efficacy and molecular mechanism of cinobufagin in non‐small cell lung cancer (NSCLC) via pyroptosis induction. Bronchial epithelial cells and NSCLC cell lines were treated with gradient concentrations of cinobufagin. Cell viability was evaluated using Cell Counting Kit‐8 (CCK‐8) assay. RNA‐sequencing was performed to identify differentially expressed genes. Lactate dehydrogenase (LDH) release was measured via cytotoxicity detection kit. Pyroptotic morphological changes were observed by transmission electron microscopy. Western blotting analysed expression levels of pyroptosis‐related proteins. In vivo efficacy was validated in nude mouse xenograft models. Immunohistochemistry evaluated tumour pyroptosis markers, whilst flow cytometry analysed tumour‐infiltrating CD8+ T cells and natural killer (NK) cells. Cinobufagin demonstrated selective cytotoxicity against NSCLC cells with minimal toxicity to normal bronchial epithelium. RNA‐seq analysis revealed significant enrichment of pyroptosis‐related pathways. Functional experiments confirmed cinobufagin‐induced LDH release, characteristic pyroptotic morphological changes and upregulation of cleaved caspase‐3 and Gasdermin E (GSDME)‐NT in NSCLC cells. In xenograft models, cinobufagin treatment reduced tumour volume compared to controls. Mechanistically, this was associated with enhanced caspase‐3 activation and GSDME‐NT accumulation in tumour tissues. Notably, cinobufagin treatment significantly increased NK cell infiltration and activity. Cinobufagin exerts antitumor effects in NSCLC through caspase‐3/GSDME‐mediated pyroptosis induction, accompanied by immune microenvironment modulation. These findings provide preclinical evidence for cinobufagin as a potential therapeutic agent targeting pyroptosis in NSCLC.

This study systematically investigated the effects of acute heat stress (AHS) on Muscovy ducks, focusing on plasma stress indicators, meat quality, myofibrillar proteins (MPs) properties, and serum metabolite profiles. AHS significantly elevated stress level, impairing meat quality, as evidenced by higher L* value, shear force, drip loss, and cooking loss (P < 0.05). It also induced MPs oxidation and aggregation, reflected by elevated carbonyl content, turbidity, zeta potential, and particle size. MPs structural alterations were confirmed by fluorescence quenching and increased exposure of hydrophobic groups. Correlation analysis revealed strong associations between physicochemical changes and MPs oxidation. Metabolomic analysis identified 161 and 105 differential metabolites in the CON vs. LS and CON vs. SS comparisons, respectively, involving 25 metabolic pathways related to energy, amino acids, and fatty acids. These findings provide novel insights into the molecular pathways of AHS-induced meat quality deterioration and reveal potential intervention timing for the poultry industry.