Combination of Zonisamide and Dexrazoxane in the Modulation of Doxorubicin-Induced Renal Damage in Rats

Abstract

Kidney injury (KI) is a significant complication of doxorubicin (DOX) chemotherapy, limiting its clinical utility. This study aimed to identify key protein targets involved in DOX-induced KI and to investigate the potential protective effects of zonisamide (ZS) and dexrazoxane (DEX), both individually and in combination, by integrating network pharmacology, molecular docking, and in vivo experimental validation. Network analysis and molecular docking to identify critical hub proteins. An in vivo experiment was subsequently done to validate the findings. The research encompassed five groups of rats: control, DOX, DEX, ZS, and DEX + ZS. Each group comprised eight rats. In vitro cytotoxicity tests, together with biochemical analyses, qPCR, Western blotting, histological evaluations, and immunohistochemistry assessments, were conducted. Network analysis identified four crucial hub proteins (Myeloid differentiation primary response 88 (MYD88), Toll-like receptor 8 (TLR8), Nuclear factor kappa B subunit 1 (NFKB1), and DNA topoisomerase IIα (TOP2A)) implicated in KI pathogenesis. In vitro cell cytotoxicity assay demonstrates a partial recovery of cell viability, particularly with the combination therapy of ZS and DEX. In a rat model of DOX-induced KI, treatment with ZS and DEX, particularly in combination, significantly ameliorated kidney dysfunction, as evidenced by reduced serum creatinine, serum cystatin c, urinary glucose, urinary albumin, and blood urea nitrogen (BUN), and attenuated histopathological damage. Moreover, both DEX and ZS restored oxidative stress markers; however, the combination therapy enhanced the responses, resulting in a further increase in superoxide dismutase (SOD)/nitrite and a greater reduction in malondialdehyde (MDA). Furthermore, ZS and DEX, alone and combined, modulated the expression of these identified hub proteins in kidney tissue, as demonstrated by qPCR, immunohistochemistry, and Western blotting, suggesting their involvement in the observed nephroprotection. The combination of ZS and DEX exhibited a protective effect against DOX-induced KI, likely because of its antioxidant and anti-inflammatory properties. Our findings underscore a potential multi-target treatment strategy for DOX-induced kidney injury, highlighting the efficacy of integrated computational approaches in drug repurposing and development.