Esculetin and Phloretin Combination Mitigates Acute Kidney Injury‐Diabetes Comorbidity via Regulating Mitophagy and Inflammation: A Dual‐Pronged Approach

ABSTRACT Induction of PINK1/Parkin‐mediated mitophagy and reducing inflammation via targeting the TLR4/NF‐κB axis simultaneously could be a promising therapy for the complex pathophysiology of AKI‐diabetes comorbidity. Earlier, esculetin by mitophagy activation and phloretin by inhibiting inflammation have shown promising renoprotection. Therefore, we aimed to evaluate the synergistic renoprotective ability of esculetin and phloretin combination against AKI‐diabetes comorbidity. AKI‐diabetes comorbidity was mimicked in vivo by bilateral ischemia/reperfusion injury (IRI) in diabetic rats and in vitro by sodium azide‐induced hypoxia/reperfusion injury (HRI) under hyperglycemic conditions. The cells were pretreated with esculetin (50 μM) and phloretin (50 μM) for 24 h. Similarly, the diabetic AKI rats received esculetin (50 mg/kg/day, p.o.) and phloretin (50 mg/kg/day, p.o.) pretreatment for 4 days and 1 h before surgery. Further, the obtained samples were utilized for different experiments. Esculetin and phloretin in diabetic AKI rats preserved kidney function and prevented kidney injury, indicated by reduced plasma creatinine, blood urea nitrogen, and kidney injury molecule 1. Esculetin improved mitophagy, indicated by increased mitophagosome formation, increased PINK1, Parkin, LC3B, and decreased p62 expression. Similarly, phloretin suppressed the diabetic AKI‐related increased expression of inflammatory mediators including NF‐κB, TLR4, TNF‐α, and MCP‐1. Moreover, combination therapy showed a more pronounced effect via synergistically improving mitophagy, maintaining ΔΨm, preventing mitochondrial dysfunction, reducing inflammation, and apoptosis. Esculetin and phloretin combination ameliorated AKI‐diabetes comorbidity more effectively than their monotherapies. Esculetin upregulated the PINK1/Parkin‐mediated mitophagy, and phloretin reduced inflammation by inhibiting the TLR4/NF‐κB axis, thereby synergistically preventing kidney dysfunction.