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This paper systematically studied the chemical constituents of Venenum , the effects of Venenum extract and monomeric compounds (Cinobufotalin and desacetylcinobufotalin) on proliferation of osteosarcoma LM-8 cell lines in C3H mice and their relationships with Bcl-2, Bax apoptin expressions were studied, and their mechanisms of apoptosis induction were preliminarily explored. Traditional Chinese medicine Venenum was isolated by silica gel column chromatography, HPLC and recrystallization, and the compound structures were elucidated using modern spectroscopic methods such as 1H-NMR and 13C-NMR, combined with physicochemical properties. WST-8 colorimetric assay was used to detect the effects of 15 μg/ml Venenum extract, Cinobufotalin and desacetylcinobufotalin on growth of LM-8 cell lines; changes in cell morphology were observed; flow cytometry was applied to detect the changes in apoptosis rate; and Bcl-2 and Bax protein expressions were detected. The structural elucidation revealed that the two compounds isolated were cinobufotalin and desacetylcinobufotalin. Desacetylcinobufotalin could significantly inhibit the growth of LM-8 cells, and the effect was dependent on concentration and time. Desacetylcinobufotalin can induce apoptosis of LM-8 cells, and the action was dose dependent, the mechanism is achieved by up-regulation of Bcl-2 expression and down-regulation of Bax expression.
Venenum , Desacetylcinobufotalin, 13C-NMR, 1H-NMR, Osteosarcoma LM-8