Network pharmacology mechanism of scutellarin involved in CASP3 and TNF for the treatment of testicular injury in diabetic mellitus
Yan Duan, Kevin FangAbstract
Objective:
This study aimed to investigate the mechanism of scutellarin (SCU) in improving testicular injury in diabetic rats by integrating network pharmacology and molecular docking, combined with experimental validation.
Methods:
Relevant targets for “scutellarin,” “diabetes mellitus,” and “testis” were collected from GeneCards, followed by intersection analysis. The STRING database was employed to construct a protein-protein interaction (PPI) network. The DAVID database was utilized for Gene Ontology (GO) biological enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses, and a “Disease-Site-Drug-Key Targets-KEGG Pathways” network was established. Molecular docking validation of SCU with CASP3, BCL2, and TNF was performed using AutoDock software. Finally, a streptozotocin (STZ)-induced type I diabetic rat model was established, and each testis was used to perform HE staining and RT-qPCR in each group.
Results:
Network pharmacology predicted a total of 53 intersecting targets for SCU acting on diabetic testicular injury, from which 10 core genes (AKT1, TP53, HIF1A, STAT3, PTEN, CASP3, EGFR, MTOR, TNF, and BCL2L1) were found. Involved biological processes included apoptosis, angiogenesis, energy metabolism, and inflammation and are significantly enriched in signaling pathways including AGE-RAGE, JAK-STAT, PI3K-AKT, HIF-1, and IL-17. Molecular docking demonstrated that SCU has a favorable binding affinity with CASP3, BCL2, and TNF. RT-qPCR validation indicated that SCU intervention significantly downregulated on the mRNA expression of CASP3 and upregulated the expression of TNF and BCL2 in the testicular tissues of diabetic rats. HE staining revealed that seminiferous tubules exhibited regular contours with orderly arranged spermatogenic epithelium in the control group, while it showed atrophy and deformation of seminiferous tubules, thinning of the spermatogenic epithelium, disordered cell arrangement, and visible vacuolation in the diabetic model group. Moreover, SCU treatment presented relatively regular contours, with increased spermatogenic epithelial thickness, improved cell arrangement, and reduced vacuolation inseminiferous tubules.
Conclusion:
SCU could exert a protective effect against diabetes-related testicular injury by interacting with core targets such as CASP3, BCL2L1, and TNF, thereby regulating signaling pathways including AGE-RAGE, PI3K-AKT, HIF-1, and TNF.