#5449 CALYCOSIN ATTENUATES RENAL ISCHEMIA/REPERFUSION INJURY BY SUPPRESSING INFLAMMATION VIA PPARγ/EGR1 PATHWAY

Abstract Background and Aims Renal ischemia reperfusion injury (IRI) is a leading common cause of AKI, delayed recovery IRI contributes to chronic kidney disease even end-stage disease. However, there no effective prevention treatment for IRI-AKI. Calycosin (CAL), an isoflavonoid phytoestrogen isolated from Radix astragali, has various pharmacological activities. whether CAL protective effect on renal its mechanism remains elusive. In this study, we aim explore the IRI-AKI through bioinformatics experiments in vitro vivo models, so as provide potential candidate drugs early intervention Method (1) first part, mice were intragastrically given daily 7 d, subjected bilateral artery occlusion 35 min followed by 24 h establish model. HE staining was used evaluate pathological injury, function including serum creatinine (SCr) blood urea nitrogen (BUN), also examined. Meanwhile, inflammatory factors detected. cell experiments, HK-2 cells pretreated with 8,16 32 μM hypoxia/reoxygenation (H/R), H/R induced investigated qRT-PCR ELISA. (2) second differentially expressed gene (DEGs) obtained analysis GSE52004 dataset. Subsequently, hub highest score protein-protein interaction (PPI) analysis, upstream transcription predicted JASPAR database. mice, results verified qRT-PCR, Western Blot immunohistochemistry. cells, regulatory relationship between factor role them process double luciferase reporter siRNA. Furthermore, molecular docking predict factor, target protein protection calycosin Results IRI, dose dependently alleviated SCr BUN levels tubular score. mRNA concentrations cytokines markedly inhibited pretreatment. pre-experiment vitro, toxicity negligible at or less incubation. incubation reduced hypoxia-inducible 1α (HIF-1α) cytokines. (3) 58 DEGs identified mice. PPI showed growth response 1 (EGR1) database that peroxisome proliferator-activated receptor γ (PPARγ) EGR1. (4) after EGR1 up-regulated PPARγ down-regulated, while pretreatment decreased increased dose-dependent manner. (5) expression siRNA, level significantly, suggesting participated promoted cells. addition, dual assay confirmed interact promoter, promoter plasmid transfection but overexpression. (6) promote markedly. According docking, CAL, ligand, could be embedded into orthosteric pocket binding energy -6.46 kcal/mol. incubated considerably. inflammation absent when knockdown, indicated ameliorate inhibit via PPARγ/EGR1. Conclusion significantly modulate targeting PPARγ/EGR1 pathway, contributing alleviation IRI.