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Punica Granatum Rind Extract Promotes Mouse Liver Mitochondrial Function and Attenuates Paraquat Toxicity in a Caenorhabditis Elegans Model

Punica granatum (pomegranate) has drawn the interest of the scientific community due to its rich array of phenolic compounds and antioxidant capacity. Evidence supports the fruit's role in preventing free radical damage in humans. Although various studies have concluded the beneficial effects of pomegranate juice, limited studies have characterized the antioxidant effects of its rind (outer covering/peel; PRE). The goal of our study was to characterize the effects of PRE on oxidative stress. We utilized mouse liver mitochondria and paraquat (PQ) toxicity in a Caenorhabditis elegans (C. elegans) model to determine the beneficial effects of PRE. Mitochondria were isolated from C57 male mice liver tissue (6 weeks of age). Phosphatidylcholine/cholesterol liposomes (Lp) were used to induce oxidative stress. Samples were then incubated at 37□ for 2 hours with or without the addition of PRE at two levels (PRE1 and PRE2 represent 60 and 120 μM Trolox equivalent/mL, respectively). Oxygen consumption rate (OCR; nmol O2 consumed/min per mg mitochondria) was used to express the activity of NADH‐CoQ oxidoreductase, succinate dehydrogenase, and cytochrome c oxidase (complex I, II, and IV of electron transport chain, respectively). Antioxidant level in PRE was determined by DPPH‐, ABTS‐ and Folin‐Ciocalteu assays. The experiment was repeated four times. Pre‐incubation of mitochondria with Lp decreased (p < 0.05) OCR by 50% compared with control mitochondria without Lp. However, incubation of Lp with PRE2 increased (p < 0.05) OCR for complex II and IV. No significant changes (p > 0.05) in OCR were observed for complex I with incubation of Lp and PRE 1 or PRE2. Using C. elegans, an oxidative stress resistance assay was performed to explore PREs effects on paraquat toxicity. Wild‐type adult/L4 hermaphrodites were placed in 96‐well plates. PQ was dissolved to a final concentration of 50 μM and half concentration of PRE1 was added for experimental groups. Resistance to oxidative stress was measured through percentage survival at hour time intervals. After 5 hours, mean survival for PQ alone was 61.91±6.56%; PQ groups supplemented with PRE had a mean survival of 88.29±6.18%. The current study suggests that PRE may play a role in preserving mitochondrial function during oxidative stress and attenuates PQ toxicity.