Ameliorative potential of pomegranate juice against synthetic colorant (E124)–induced biochemical and histopathological changes in the rat brain and testes

The present study investigated the biochemical and molecular interactions between the synthetic azo dye ponceau 4R (E124) and the bioactive constituents of natural pomegranate ( Punica granatum ) juice. We hypothesized that the rich profile of polyphenols and anthocyanins in pomegranate juice could mitigate the oxidative and structural damage induced by Ponceau 4R administration. Rats were divided into four groups: control, ponceau 4R (0.7 mg/kg), pomegranate juice (1 mL/rat), and a combined treatment group. Biochemical analyses revealed that ponceau 4R significantly elevated serum markers of hepatic and renal dysfunction, disrupted lipid metabolism, and induced oxidative stress, as evidenced by increased malondialdehyde (MDA) and depleted antioxidant defenses. RT-qPCR analysis demonstrated a marked upregulation of the pro-apoptotic gene caspase-3 in ponceau 4R–treated rats. Histopathological examination of the brain and testes confirmed pronounced structural alterations consistent with these molecular findings. In contrast, supplementation with pomegranate juice ameliorated these adverse effects by enhancing the antioxidant status, downregulating caspase-3 expression, and preserving tissue architecture. These findings support our hypothesis that the dietary bioactive compounds in pomegranate juice exert significant protective effects against synthetic colorant–induced molecular damage. This study highlights the potential of valorizing natural food sources to enhance food safety and mitigate the risks associated with synthetic additives. • Ponceau 4R at 0.7 mg/kg induces oxidative damage in rat brain and testes • Pomegranate juice restores SOD and CAT while reducing MDA levels. • Molecular analysis shows PJ down-regulates Caspase-3 to inhibit apoptosis. • PJ mitigates E124-induced lesions, recovering organ architecture.