Renal protective efficiency of Isatin-linked Pyrazole (3E) derivative which mitigates gentamicin-induced nephrotoxicity and its anti-fibrotic mechanisms in an in-vivo zebrafish model

Gentamicin (Gen), a commonly used aminoglycoside antibiotic, is associated with significant nephrotoxicity driven by oxidative stress, inflammation, and fibrosis, leading to renal impairment. This study investigates the therapeutic potential of the Isatin-linked Pyrazole (3E) derivative (or compound) in mitigating Gen-induced nephrotoxicity using in-vivo zebrafish as a model organism. The anti-inflammatory, antioxidant, and anti-fibrotic properties of the 3E derivative were assessed through biochemical, molecular, and histopathological analyses. The 3E compound demonstrated concentration-dependent anti-inflammatory activity, significantly reducing nitric oxide production, proteinase activity, and hemolysis and maintaining the safest dosages in zebrafish embryos. Treatment with 3E effectively restored antioxidant enzyme levels (SOD, CAT, and GST) and reduced oxidative damage markers, including LDH, compared to the Gen-treated group. Additionally, 3E ameliorated Gen-induced glomerular filtration damage and collagen deposition in kidney tissues, as evidenced by reduced hydroxyproline levels and creatinine and urea excretion rate. Molecular analyses revealed that 3E significantly downregulated pro-inflammatory (il-1β, tnfα) and pro-fibrotic (tgf-β1a, mmp9) genes, correlating with histological improvements in glomerular and tubular architecture. Furthermore, the model limitations in translating results to human physiology, the findings highlight 3E's therapeutic potential for nephroprotection by targeting key mechanisms underlying renal injury. Future studies are warranted to elucidate the molecular pathways and validate the efficacy of 3E in mammalian models.