6-Shogaol Reduces Renal Macrophage Infiltration by Targeting the STING Pathway to Alleviate Cisplatin Induced Renal Injury.

Abstract

Cisplatin is a widely used chemotherapeutic agent, but its dose-limiting nephrotoxicity remains a major clinical challenge. 6-Shogaol, a bioactive compound with potent anti-inflammatory and antioxidant properties, exhibits strong binding affinity for STING-a key mediator in cisplatin-induced kidney injury. This study aims to investigate the renoprotective effects of 6-shogaol against cisplatin nephrotoxicity and elucidate its underlying mechanisms. Using a murine model of cisplatin-induced nephrotoxicity and in vitro experiments with HK-2 cells, we evaluated the cytoprotective effects of 6-shogaol. Molecular docking simulations confirmed its high binding affinity for STING. A comprehensive approach, including flow cytometry, RNA sequencing, ELISA, qPCR, histopathological staining (H&E staining), immunofluorescence, and Western blot analyses, was employed to assess renal function, tubular injury, inflammatory responses, and molecular mechanisms. CRISPR/Cas9-mediated STING knockout was performed to validate the mechanistic role of STING. 6-Shogaol significantly attenuated cisplatin-induced renal damage, as evidenced by reduced serum creatinine and blood urea nitrogen levels, diminished macrophage infiltration, and downregulated expression of CCL2 and CCL5 in renal tissues. Molecular docking revealed stable binding between 6-shogaol and STING. In vitro, 6-shogaol mitigated cisplatin-induced cellular injury, suppressed the cGAS-STING pathway, and reduced pro-inflammatory cytokine (IL-1β, IL-6, TNF-α) and chemokine (CCL2, CCL5) expression. STING knockout in HK-2 cells abolished cisplatin-induced cytotoxicity and attenuated the protective effects of 6-shogaol. These findings demonstrate that 6-shogaol alleviates cisplatin-induced kidney injury by targeting STING, thereby inhibiting CCL2/CCL5-mediated macrophage infiltration and subsequent inflammation.