GAS5 mediates notch signaling to exacerbate calcium oxalate-induced renal injury through the NF-κB/NLRP3 axis by promoting macrophage polarization.

The contribution of macrophage polarization dysregulation to calcium oxalate (CaOx) nephrolithiasis remains poorly defined. Here, we identify the long non-coding RNA growth arrest-specific transcript 5 (GAS5) as a critical driver of pro-inflammatory M1 macrophage polarization through the Notch/NF-κB/NLRP3 axis, thereby aggravating CaOx-induced renal injury. In human macrophages, calcium oxalate monohydrate (COM) stimulation upregulates GAS5 expression, which functions as a competing endogenous RNA by sequestering miR-449c-5p, leading to derepression of Notch1 and amplification of downstream inflammatory signaling. Activated Notch1 engages TLR4 to trigger NF-κB/NLRP3 inflammasome activation, promoting M1 polarization, inflammatory cytokine release, tubular epithelial pyroptosis, and enhanced crystal adhesion. Given the limited conservation of lncRNA-miRNA interactions across species, we further delineated a species adaptive regulatory mechanism in mice. In murine macrophages, GAS5 enhances Notch1 signaling through sequestration of a distinct miRNA, miR-325-3p, thereby preserving the core ceRNA logic while employing different miRNA mediators. In vivo, macrophage enriched GAS5 overexpression exacerbates Gly induced CaOx deposition, renal dysfunction, and inflammation, whereas pharmacological inhibition of Notch signaling or TLR4 knockdown markedly attenuates these effects. Conversely, restoration of the corresponding regulatory miRNAs suppresses Notch1 signaling and favors anti-inflammatory macrophage polarization. Together, these findings uncover a conserved, yet species-adaptive, GAS5 centered ceRNA regulatory framework that governs macrophage driven inflammation and renal injury in CaOx nephrolithiasis, highlighting GAS5-Notch signaling as a potential immunomodulatory target for nephrolithiasis.