The Therapeutic Potential of Chi Shao and Fu Zi in Acute-on-Chronic Liver Failure: Insights from Network Pharmacology and Animal Studies

Background & Aims Acute-on-chronic liver failure (ACLF) has a high incidence and mortality rate, which has become an urgent clinical problem to be solved.Traditional Chinese medicine (TCM) has been widely applied in the clinical and experimental researches of ACLF. But its specific mechanism is not yet fully understood. This study attempted to identify the effective active ingredients, key targets, and biological mechanisms of Chinese herbal medicines Chi Shao ( Paeoniae radix Rubra) and Fu Zi ( Aconitum carmichaelii Debx) (CSFZ) for treating ACLF based on network pharmacology and animal experiments. Methods & Results Network pharmacology was performed to predicte the major components of CSFZ and potential signal pathways. Molecular docking method was applied to determine the binding affinity between the main components and the key targets. A rat model of ACLF was constructed. The liver injury were measured by histopathological methods. Biochemical indexes were used to evaluate liver function. ELISA was used to detect the IL-6, TNF-α, Caspase-3, BCL-2, ALB, and TB level. Also, PI3K/Akt signaling molecules were detected through RT-qPCR and WB. Immunofluorescence staining was used for PCNA and Cyclin B1. In the results we found that forty-nine potential active ingredients and 104 drug targets were screened from CSFZ, and 2730 genes were identified as ACLF targets, of which 72 overlapped with CSFZ targets. we identified ten key active ingredients of CSFZ. Moreover, Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that CSFZ modulated ACLF may through numerous biological pathways, including antibacterial and anti-inflammatory mechanisms, cell proliferation, and apoptosis. Molecular docking verification shows that key components and core targets have good binding affinity. Also, through the vivo studies we found that CSFZ enhanced hepatic regeneration, improved hepatic structural damage, reduced hepatocyte apoptosis, and promoted the expression of PI3K and AKT. Conclusions Network pharmacology and in vivo experiments indicated that CSFZ may ameliorate hepatic injury by regulating the PI3k/Akt signaling pathway.