Current Pharmaceutical Design - Volume 31, Issue 20, 2025

Liquidambaris Fructus (LF), a prevalent Chinese medicinal herb, has been effectively utilized in the clinical treatment of rheumatoid arthritis (RA). Coniferin, is the active ingredient in LF, and there is a paucity of research examining its potential anti-RA properties. This study employs in silico analysis and experimental validation to delve into the therapeutic potential of Coniferin against RA and to elucidate its mechanism of action.

In silico analysis was employed to construct a drug-disease target protein-protein interaction (PPI) network, to perform functional enrichment analysis, and to molecular docking of the principal compounds and target proteins. Subsequently, the effects of coniferin on the proliferation, migration, and invasion of rheumatoid arthritis - Fibroblast-like synoviocytes (RA-FLSs) were observed using a CCK8 assay and Transwell assay. ELISA was employed to detect the inflammatory response of RA-FLSs in coniferin. Flow cytometry was utilized to detect the effects of coniferin on apoptosis, oxidative stress, and mitochondrial transmembrane potential in RA-FLSs. Ultimately, the expression of pivotal proteins and apoptosis markers within the PTGS2/Apoptosis signaling pathway was discerned through the utilization of Real-time quantitative PCR (RT-qPCR) and Western blot.

It was observed that coniferin promotes apoptosis of RA-FLSs through the PTGS2/Apoptosis signaling pathway and inhibits the proliferation, migration, and invasion of RA-FLSs, with anti-inflammatory, oxidative stress-reducing, and mitochondrial transmembrane potential disruption effects.

The potential mechanism of coniferin for the treatment of RA is to promote apoptosis of RA-FLSs by intervening in the PTGS2/apoptosis signaling pathway.

The present study delves into the exploration of diagnostic biomarkers linked with ferroptosis in the context of diabetic nephropathy, unraveling their underlying molecular mechanisms.

In this study, we retrieved datasets GSE96804 and GSE30529 as the training cohort, followed by screening for Differentially Expressed Genes (DEGs). By intersecting these DEGs with known ferroptosis-related genes, we obtained the differentially expressed genes related to ferroptosis (DEFGs). Subsequently, Weighted Correlation Network Analysis (WGCNA) was carried out to identify key modules associated with Diabetic Nephropathy (DN), culminating in the identification of a significant gene. Enrichment analysis and Gene Set Enrichment Analysis (GSEA) were then carried out on the DEFGs and genes linked to the significant gene. To validate our findings, we employed cohorts GSE30528 and GSE43950, utilizing ROC curve analysis to assess diagnostic efficacy for DN, as measured by the area under the curve (AUC). Immune cell infiltration was analyzed and compared between groups using the CIBERSORT algorithm. Bayesian co-localization analysis was performed to examine the co-location of DEFGs and DN. Finally, to validate the hub genes identified, we conducted quantitative real-time polymerase chain reaction (qRT-PCR) experiments in vitro.

FUZ, GLI1, GLI2, GLI3, and DVL2 were identified as the hub genes. Functional enrichment analysis demonstrated that ferroptosis and immune response play an important role in DN. ROC analysis showed that the identified genes had good diagnostic efficiency in DN. The results of the immune infiltration analysis showed that there may be crosstalk between ferroptosis and immune cells in DN. Bayesian co-localization analysis revealed the genetic correlation between the hub genes and DN. The outcomes of the qRT-PCR analyses corroborated the reliability of the identified hub genes as robust molecular markers for targeted therapy in DN.

The interplay between immune inflammatory reactions and ferroptosis emerges as a crucial pathogenic mechanism, offering novel insights into the molecular therapy of DN. Furthermore, the identification of FUZ, GLI1, GLI2, GLI3, and DVL2 as potential targets holds promise for future therapeutic interventions aimed at treating DN.