Assessment of toxicity and anticancer activity of attenuatumione D from Hypericum sampsonii hance against hepatocellular carcinoma.

This study investigates the anticancer effects of attenuatumione D, a bioactive compound derived from Hypericum sampsonii Hance, and its potential to inhibit the proliferation of hepatocellular carcinoma (HCC). For this purpose, we employed an in vitro study to assess the cytotoxicity of attenutumione D by measuring cell viability, oxidative stress, and pro-inflammatory cytokine levels using ELISA, as well as apoptosis using flow cytometry and Western blot in HepG2 cells. While in vivo analysis was carried out using H22-tumor-bearing nude Balb/C mice model. Our data from an in vitro study showed cytotoxicity of attenuatumione D, exhibiting time & dose-dependent variation in cell morphology and a reduction in viability of HepG2 cells, with an IC50 value of 25 μM. It was found that the anti-proliferative effects of attenuatumione D were associated with apoptotic cell death, indicated by increased early apoptosis and up-regulation of cleaved caspase-3, with increased ratios of Bax/Bcl-2. Moreover, attenuatumione D treatment reduced the pro-tumorigenic cytokines, i.e., IL-1β, IL-6, and TNF-α in HepG2 cells. These findings were concomitant with increased oxidative stress. In vivo, attenuatumione D demonstrated therapeutic capacity in H22-tumor-bearing mice by inhibiting the tumor growth and protecting the liver histology with minimal hepatic alterations. Metabolomic analysis of H22-tumor-bearing mice serum and its mechanistic validation demonstrated that attenuatumione D treatment reduced the elevated levels of inosine, hypoxanthine, and guanine (key metabolites in purine metabolism) and modulated the expression of key apoptotic proteins, i.e., JNK, ERK, and p38 associated with MAPK signaling. Collectively, this study provides pioneering evidence of the anticancer potential of attenuatumione D against HCC.