Network Pharmacology Prediction and Experimental Validation of Ferulic Acid's Protective Effects against Diclofenac-Induced Liver Injury

Despite being one of the most consumed analgesics worldwide, liver injury is an adverse effect of diclofenac (DF). In pursuit of reliable hepatoprotective natural remedies, this study aimed to investigate the potential protective effect of ferulic acid (FA) and its mechanism against DF-induced liver injury. Various network databases and datasets were used to collect targets corresponding to FA and DF-induced liver injury. Enrichment analyses of common targets were performed, a protein-protein interaction (PPI) network was constructed, the hub genes were identified, and the upstream miRNA interacting with the top hub gene was later predicted. A DF-induced liver injury rat model was established to verify FA's protective effects, and the selected hub gene expression level with its upstream regulatory miRNA and a downstream set of targets was examined to elucidate the underlying mechanism. A total of 18 genes were identified as potential targets of FA to protect against DF-induced liver injury. Data from the enrichment and PPI analyses and the prediction of the upstream miRNAs indicated that the most worthwhile pair to study was miR-296-5p/Jun. In vivo findings showed that coadministration of FA significantly reduced the DF-induced alterations in the liver function indices, oxidative stress, and liver histology. Mechanistically, FA downregulated the expression of Jun, Bim, Bax, Casp3, IL-1 beta, IL-6, and TNF-alpha, whereas it upregulated the expression of rno-miR-296-5p and Bcl2. In conclusion, combining network pharmacology and an in vivo study revealed that miR-296-5p/Jun axis could mediate the mitigative effect of FA against DF-induced liver injury.