As the principal bioactive constituents of Venenum Bufonis (VB), bufadienolides mediate the therapeutic efficacy of this traditional Chinese medicine in diverse clinical applications. In this study, a novel bufadienolide, designated as bufovende A (1), and seven known analogs (2-8) were isolated from VB. The structure of bufovende A was determined using spectroscopic methods (1D/2D NMR and HRESIMS). Given the pivotal role of hepatic stellate cell (HSC) activation in liver fibrogenesis, we evaluated the antifibrotic potential of these compounds using the LX-2 human HSC line. The results indicated that non-cytotoxic concentrations (0.2 μM) of bufadienolides significantly suppressed the mRNA expression of fibrosis biomarkers α-SMA and COL1A1, with bufovende A exhibiting the most pronounced inhibitory efficacy (inhibition rates of 61.33% ± 1.54% for α-SMA and 57.46% ± 12.03% for COL1A1). Additionally, compared to the model group, bufovende A downregulated α-SMA, COL1A1, and TGF-β protein expression. These results highlight the potential of bufadienolides as promising antifibrotic leads, with bufovende A being particularly promising for further investigation.
