Abstract: Objective　To investigate the effect of Fuzheng Huayu (Fzhy) Formula drug-containing serum on the activation of activinA/smad signaling pathway in hepatic stellate cells (HSC). Methods　Twenty SD rats were divided into the control group and the Fzhy-low, medium and high dose drug-containing serum group according to the random number table method, 5 rats in each group. Rats were gavaged with distilled water, 0.75, 1.5 and 3.0 g/kg Fzhy solution (drug powder of Fzhy capsule prepared with distilled water) once/d for 3 days, and blood samples were collected to prepare blank serum (the control group) and the drug-containing serum. HSC-T6 cells were cultured with 5%, 10% and 20% of the drug-containing serum in different volume fractions, respectively. Flow cytometry was used to detect cell cycle and apoptosis rate, mitochondrial membrane potential changes and reactive oxygen species (ROS) levels. Real-time fluorescence quantitative polymerase chain reaction was used to detect mRNA levels of activinA, smad3, samd7 and nuclear factor (NF)-κB in cells. Protein blotting was used to detect activin ⅡA receptor (ActRⅡA), smad3, NF-κB p65, and cysteine aspartate protease (caspase)-3 protein levels. Results　The cell survival rate of each Fzhy -containing serum group was lower than that of the control group (P＜0.05), and the drug-containing serum with a volume fraction of 10% was selected for subsequent experiments. There were no significant differences in cell cycle and apoptosis rates between the control group and each Fzhy-containing serum group. The proportion of intracellular ROS level and mitochondrial membrane potential level reduction increased in the control group and the Fzhy-low, medium and high dose groups successively (P＜0.05). Compared with the control group, smad3 mRNA expression level was decreased, and smad7 mRNA was increased in the Fzhy-medium and high dose groups. NF-κB and activinA mRNA, smad3, NF-κB p65 and ActRⅡA protein expression levels were decreased in the Fzhy-low and medium and high dose groups, and caspase-3 protein expression levels in the Fzhy-low and medium dose groups were increased (P＜0.05). Compared with the Fzhy-low dose group, smad3 mRNA expression levels were decreased in the Fzhy-medium and high dose groups, and activinA and smad7 mRNA were increased in the Fzhy-medium dose group (P＜0.05). Conclusion　Fzhy containing serum can achieve antifibrotic effects by affecting the proliferation of HSC-T6 cells, participating in cellular oxidative stress, and regulating cellular activinA/smad signaling pathways.

Key words: liver cirrhosis, hepatic stellate cells, Smad proteins, reactive oxygen species, activins, activin receptors, type Ⅱ, Fuzheng Huayu recipe