Objective: To explore the role and mechanism of hyperforin (one of the active components of Sophora flavescens) in renal fibrosis.
Methods: The active compounds and target proteins of Sophora flavescens were first screened through TCMSP (https://tcmsp-e.com/). The renal fibrosis-related genes were analyzed through GeneCards (https://www.genecards.org/). The differentially expressed genes (DEGs) in renal fibrosis in GEO dataset GSE156181 were obtained. Metascape was applied for target protein enrichment analysis. TGF-β1-stimulated renal tubular epithelial cells were used for renal fibrosis cell model establishment. The unilateral ureteral obstruction (UUO) mouse model was used for the renal fibrosis in vivo model. Cell viability was detected using an MTT assay. Immunofluorescence staining was employed to detect cell morphology changes and the expression of α-SMA and collagen I. Hematoxylin and eosin (H&E) and Masson staining were employed to determine the renal morphologic change. qRT-PCR or Western blotting was applied to determine the expression levels of the target proteins.
Results: After intersecting the analysis results of TCMSP, GeneCards, and dataset GSE156181, hyperforin targeting ICAM1 was identified. Metascape pathway enrichment analysis results revealed that the effective compounds of Sophora flavescens were tightly associated with extracellular matrix (ECM) remodeling and inflammatory response. MTT assay demonstrated that hyperforin had no toxic effect on cells. Immunofluorescence staining results evidenced that hyperforin could partially restore TGF-β1-induced epithelial-mesenchymal transition (EMT), the PI3K/AKT pathway activation, and ICAM1 upregulation, and these effects of hyperforin could be reversed by ICAM1 overexpression. While the PI3K/AKT pathway activator IGF-1 effectively reversed the EMT inhibition effect of hyperforin on renal tubular epithelial cells. Moreover, the UUO mouse model further confirmed that hyperforin reduced renal fibrosis.
Conclusion: Hyperforin inhibited renal fibrosis via the PI3K/AKT/ICAM1 axis.
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