Regulation of Bcl-2 family molecules and activation of caspase cascade involved in gypenosides-induced apoptosis in human hepatoma cells

Herbal medicines are increasingly being utilized to treat a wide variety of disease processes. Gypenosides (Gyp) are triterpenoid saponins contained in an extract from Gynostemma pentaphyllum Makino and reported to induce apoptosis in human hepatoma cells. However, the molecular mechanism underlying the Gyp-induced apoptotic process is unclear. In this study, we found that Gyp induced apoptosis in human hepatoma Huh-7, Hep3B and HA22T cell lines as evidenced by morphological changes, 4',6'-diamidino-2-phenylindole staining and in situ terminal transferase-mediated dUTP-fluorescensin nick end-labeling assay. Our data demonstrated that Gyp-induced apoptotic cell death was accompanied by up-regulation of Bax, Bak and Bcl-X-L, and down-regulation of Bcl-2 and Bad, while it had no effect on the level of Bag-1 protein. Moreover, Gyp treatment caused the release of mitochondrial cytochrome c to cytosol and sequential activation of caspases, including caspase-1, -9 and -3, then leading to cleavage of poly-ADP-ribose polymerase. Furthermore, the Gyp-induced apoptosis was markedly blocked by the broad-spectrum caspase inhibitor, z-VAD-fmk. Taken together, these results suggest that treatment of human hepatoma cells with Gyp induced apoptosis through the up-regulation of Bax and Bak, and down-regulation of Bcl-2, release of mitochondrial cytochrome c and activation of caspase cascade. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.