In vivo and in vitro evaluation of antioxidant, antihemolytic, acute toxicity effects and high-performance liquid chromatography analysis of an aqueous extract of Moroccan Punica granatum L.

Introduction: The use of pomegranates in Moroccan pharmacopeia is due to their healing and nutritional properties because of their richness in secondary molecules. The following study analyses the composition of the aqueous extract of Punica granatum peel and evaluates in vivo and in vitro antioxidant effects, hemolytic protection, and acute toxicity. Methods: Quantification of the plant extract was realized by high-performance liquid chromatography (HPLC). The hemolytic assay was used for erythrocyte protection, while the in vitro antioxidant effect was evaluated by ۲, ۲۰-azinobis-(۳-ethylbenzothiazoneline-۶-sulphonic acid) (ABTS) and reducing ferric power (RFP) assays. The in-vivo antioxidant activity was tested by measuring levels of lipid peroxidation (LPO) in serum. The toxicological study was tested by oral administration of the extract to four groups of mice for ۲۱ days, followed by a histopathological examination of the spleen. Results: HPLC analysis showed the presence of some phenolic compounds such as coumarin, caffeic, gallic and syringic acids. The IC۵۰ of the antioxidant assays were ۲۵۴.۴۹ ± ۶۲.۱۷ μg/mL and ۴۰.۲۶۵ ± ۲.۹ μg/mL for ABTS and reducing power, respectively. Furthermore, the thiobarbituric acid reactive substances (TBARS) assay showed the lowest levels at ۱۵۰ mg/mL concentration. All of the concentrations used for hemolytic protection did not exceed ۱۵% of hemolysis. Moreover, the toxicity test showed no sign of mortality, signs of weakness, or weight loss; also the histopathological examination of the spleen tissues showed the absence of any damage. Conclusion: The peel extract of P. granatum showed good potential and could be exploited as a natural antioxidant and antihemolytic remedy, leading to the development of new drugs.

Keywords:

Pomegranate peel , Herbal medicine , Lipid peroxidation , Polyphenols , Animal experiments