OBJECTIVE  To screen traditional Chinese medicines with potential anti-fibrotic effects and explore their anti-renal fibrosis effects and mechanisms.

METHODS  By integrating resources from the OMIM, GeneCards, TTD, and TCMSP databases, traditional Chinese medicines with potential anti-fibrotic effects were systematically screened. Unilateral ureteral obstruction(UUO) mouse model was used to evaluate the intervention effects of target traditional Chinese medicine on renal fibrosis. Anti-renal fibrosis effect of target traditional Chinese medicine on primary renal fibroblasts(PRFs) model was further examined, and the technique tandem mass tag(TMT) was used to detect differentially expressed proteins(DEPs) and bioinformatics analysis was performed. Key proteins were screened by both probable protein-protein interactions network and literature search, followed by the next step of cellular experimental validation.

RESULTS  Network pharmacology analysis revealed that Ephedrae Herba exhibited significant anti-fibrotic potential. In vivo experiments confirmed that Ephedrae Herba effectively improved renal pathological damage and inhibited the progression of renal fibrosis. In vitro experiments further validated its anti-fibrotic effects at the cellular level. Through TMT quantitative proteomics analysis, among the 4 973 proteins detected, 65 significant DEPs were identified, with FN1, MDA5, MX1, ALB, and ISG15 identified as key regulatory proteins. Cellular experiment validation results showed that after Ephedrae Herba intervention, the protein expression levels of FN1, MDA5, MX1, ALB, and ISG15 were significantly downregulated, consistent with the expression trends observed in TMT quantitative proteomics analysis.

CONCLUSION  This study systematically elucidates for the first time that Ephedrae Herba exhibits significant anti-renal fibrosis effects in both in vivo and in vitro experimental models and identified FN1, MDA5, MX1, ALB, and ISG15 as potential key molecular targets for Ephedra’s anti-renal fibrosis effects.