Astragaloside IV Alleviates Osteoarthritis by Upregulating ETS2: A Bioinformatics and Experimental Study

Osteoarthritis (OA) is a common degenerative joint disorder and there are currently no effective therapies to impede its destructive progression. Astragaloside IV (AS-IV), a natural compound, exhibits promising chondroprotective effects, yet its specific molecular mechanisms remain poorly clarified. Thus, this study employed transcriptomic profiling combined with bioinformatics analysis to identify OA-related characteristic genes, and further conducted experiments to verify the potential therapeutic mechanism of AS-IV.

Analysis of the GSE114007 dataset was performed using False Discovery Rate (FDR)-adjusted p-values and quantile normalization to mitigate batch effects. Weighted gene co-expression network analysis (WGCNA) was utilized to identify key modules. Nine OA-related feature genes were identified and validated in two external datasets (GSE129147 and GSE51588) using LASSO with 10-fold cross-validation and a random forest algorithm to minimize the risk of optimistic bias. Furthermore, gene set enrichment analysis (GSEA) was conducted. The underlying mechanisms were validated through molecular docking, cellular thermal shift assay (CETSA), as well as in vitro experiments using ATDC-5 cells and in vivo assays with OA mice.

We identified 1548 differentially expressed genes (DEGs) primarily enriched in the extracellular matrix, with PI3K-Akt signaling closely related to OA. The MEBlack module was strongly associated with OA (cor = -0.79, p<0.0001). GSEA showed the feature genes were associated with the adipocytokine signaling pathway and glycosaminoglycan biosynthesis. Furthermore, molecular docking and CETSA indicated that ETS2 served as a potential interacting target of AS-IV. In the destabilization of the medial meniscus (DMM)-induced OA mice, the results of Safranin O/Fast Green staining confirmed that AS-IV alleviated cartilage loss and lowered OARSI scores. Mechanistically, AS-IV slowed OA progression by upregulating Col2a1 expression, suppressing MMP13 levels, and reducing inflammatory markers such as IL-1β and TNF-α. Importantly, AS-IV enhanced ETS2 expression in osteoarthritic chondrocytes. Consistently, in vitro functional assays revealed that knockdown of ETS2 in chondrogenic ATDC-5 cells partially reversed both the chondroprotective and anti-inflammatory effects of AS-IV, verifying the essential role of ETS2 in mediating the therapeutic effects of AS-IV against OA.

Collectively, these findings point to the vital function of ETS2 in OA pathogenesis and demonstrate that AS-IV attenuates cartilage degradation and inflammatory responses by upregulating ETS2, thereby retarding OA development.