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- Title
扶正化瘀方含药血清对肝星状细胞 activinA/smad 信号通路活化的影响.
- Authors
陈黎旭; 熊佳; 谢昆; 朱挺德; 钟志英; 官亮; 潘永平
- 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 drugcontaining 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 Fzhycontaining 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 ActR Ⅱ A 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.
- Publication
Tianjin Medical Journal, 2022, Vol 50, Issue 7, p707
- ISSN
0253-9896
- Publication type
Academic Journal
- DOI
10.11958/20220030