Mechanism Of Rhizoma Corydalis In Prevention And Treatment Of Liver Fibrosis Was Researched By Regulating Ferroptosis Based On PI3K/Akt/P53 Signal Axis

Objective The effects of Rhizoma Corydalis（RC）on Hepatic fibrosis（HF）were investigated by network pharmacology and animal experiments.Methods 1.Using the network pharmacological approach,“RC”,“liver fibrosis”and“hepatic fibrosis”were searched in relevant databases as keywords.The Gene Ontology（GO）and Kyoto Encyclopedia of Genes and Genomes（KEGG）were obtained using the Microbiotics online tool.The corresponding"drug-component-target-pathway"network diagram was constructed to obtain the possible targets and pathways.2.The rat model of liver fibrosis induced by CCl ₄was established,and the liver function of the rats with liver fibrosis was evaluated by serum biochemical analysis.The liver fibrosis indexes were determined by histopathological analysis,immunohistochemical analysis and enzyme-linked immunosorbent assay to evaluate the degree of liver fibrosis.Ferroptosis index in liver tissu e was detected by relevant kit.The expression of PI3K,Akt,P53,GPX4 and PTGS2 genes was determined by RT-PCR.The protein expressions ofα-SMA,p-PI3K,p-Akt,p-P53,GPX4 and PTGS2 were detected by western blotting technique.The ferroptosis index and s ignaling pathway of RC treatment on carbon tetrachloride induced hepatic fibrosis rat model were investigated in vivo.Results 1.In this paper,we obtained 49 main components of RC such as quercetin,berberine and dehydroaconitine through network pharmacology screening,168remaining after removing duplicate target genes,1126 liver fibrosis-related target genes,80 potential action targets such as Akt,PTGS2 and P53 of RC for liver fibrosis by topological analysis;The analysis of KEGG yielded 171signaling pathways,which mainly included PI3K/Akt signaling pathways.2.Based on the analysis of network pharmacology,we demonstrated the anti-liver fibrosis effect of RC,and constructed a CCl₄-induced liver fibrosis model in rats.The results showed that compared with the CCl₄ group,the RC administration group significantly reduced the serum alanine transaminase（ALT）,aspartate aminotransferase（AST）,alkaline phosphatase（ALP）and total bile acid（TBA）.The levels of hydroxyproline（HYP）,hyaluronic acid（HA）and laminin（LN）were reduced.Then,HE and Masson staining results showed that the RC treatment groups significantly reduced inflammatory cell infiltration,fat vacuoles and collagen deposition compared with the model group;immunohistochemical analysis showed that the RC treatment group reduced the levels of alpha-SMA（α-SMA）and Collagen-Ⅰin liver tissue.In addition,Fe²⁺,lipid peroxidation（LPO）and malondialdehyde（MDA）increased and superoxide dismutase（SOD）decreased in the RC administration groups.Ultimately,the mRNA levels in liver tissues were determined.Compared with CCl₄ group,the gene levels of PI3K,Akt and GPX4 in RC treatment groups were decreased,while the gene levels of P53 and PTGS2 were increased.WB experiment results showed that compared with CCl₄group,the protein levels of p-PI3K,p-Akt and GPX4 were decreased with RC treatment,while the protein levels of p-P53 and PTGS2 were increased.These results suggested that RC was effective in preventing and treating liver fibrosis.Conclusion These results suggested that RC might play a role in anti-liver fibrosis by regulating the PI3K/Akt/P53 signaling axis to promote ferroptosis.