Calycosin inhibited MIF-mediated inflammatory chemotaxis of macrophages to ameliorate ischemia reperfusion-induced acute kidney injury

• Published in: Inflammation research 2024-08, Vol.73 (8), p.1267-1282
• Main Authors: Wang, Hong-Lian, Peng, Ze, Li, Yu-Qing, Wang, Yi-Xuan, Li, Jian-Chun, Tan, Rui-Zhi, Su, Hong-Wei, Shen, Hong-Ping, Zhao, Chang-Ying, Liu, Jian, Wang, Li
• Format: Article
• Language: English
• Subjects: Acute Kidney Injury - drug therapy; Acute Kidney Injury - metabolism; Allergology; Animals; Anti-Inflammatory Agents - pharmacology; Anti-Inflammatory Agents - therapeutic use; Attenuation; Biomedical and Life Sciences; Biomedicine; Cell activation; Chemokines; Chemotaxis; Chemotaxis - drug effects; Creatinine; Dermatology; Down-regulation; Immunology; Infiltration; Injury analysis; Intramolecular Oxidoreductases - genetics; Intramolecular Oxidoreductases - metabolism; Ischemia; Isoflavones - pharmacology; Isoflavones - therapeutic use; Kidney - drug effects; Kidney - pathology; Kidney diseases; Kidneys; Leukocyte migration; Lipopolysaccharides; Macrophage migration inhibitory factor; Macrophage Migration-Inhibitory Factors; Macrophages; Macrophages - drug effects; Male; Mice; Mice, Inbred C57BL; Molecular docking; Monocyte chemoattractant protein 1; Neurology; NF-kappa B - metabolism; NF-κB protein; Original Research Paper; Pharmacology; Pharmacology/Toxicology; RAW 264.7 Cells; Reperfusion; Reperfusion Injury - drug therapy; Rheumatology; Ribonucleic acid; RNA; Surface plasmon resonance; Target spot; Tumor necrosis factor-α; Urea
• ISSN: 1023-3830; 1420-908X; 1420-908X
• DOI: 10.1007/s00011-024-01899-0

Background Inflammatory macrophage infiltration plays a critical role in acute kidney disease induced by ischemia-reperfusion (IRI-AKI). Calycosin is a natural flavone with multiple bioactivities. This study aimed to investigate the therapeutic role of calycosin in IRI-AKI and its underlying mechanism. Methods The renoprotective and anti-inflammatory effects of calycosin were analyzed in C57BL/6 mice with IRI-AKI and lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. RNA-seq was used for mechanism investigation. The molecular target of calycosin was screened by in silico methods and validated by surface plasmon resonance (SPR). Macrophage chemotaxis was analyzed using Transwell and agarose gel spot assays. Results Calycosin treatment significantly reduced serum creatinine and urea nitrogen and attenuated tubular destruction in IRI-AKI mice. Additionally, calycosin markedly suppressed NF-κB signaling activation and the expression of inflammatory mediators IL-1β and TNF-α in IRI-AKI kidneys and LPS-stimulated RAW 264.7 cells. Interestingly, RNA-seq revealed calycosin remarkably downregulated chemotaxis-related pathways in RAW 264.7 cells. Among the differentially expressed genes, Ccl2/MCP-1, a critical chemokine mediating macrophage inflammatory chemotaxis, was downregulated in both LPS-stimulated RAW 264.7 cells and IRI-AKI kidneys. Consistently, calycosin treatment attenuated macrophage infiltration in the IRI-AKI kidneys. Importantly, in silico target prediction, molecular docking, and SPR assay demonstrated that calycosin directly binds to macrophage migration inhibitory factor (MIF). Functionally, calycosin abrogated MIF-stimulated NF-κB signaling activation and Ccl2 expression and MIF-mediated chemotaxis in RAW 264.7 cells. Conclusions In summary, calycosin attenuates IRI-AKI by inhibiting MIF-mediated macrophage inflammatory chemotaxis, suggesting it could be a promising therapeutic agent for the treatment of IRI-AKI.