Characterization of a pomegranate (Punica granatum) extract towards development of phenolic acids based on a non-opioid substitution therapy: in vitro and in vivo studies

Mohd Adzim Khalili, Rohin and Atif Amin, Baig and Mimie Noratiqah, Jumlia and Norhaslinda, Ridwana (2018) Characterization of a pomegranate (Punica granatum) extract towards development of phenolic acids based on a non-opioid substitution therapy: in vitro and in vivo studies. Health and the Environment Journal, 9 (1). b14-b27. ISSN 2180-1126

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Abstract

Pomegranate is a nutrient dense fruit rich in phytochemical compounds, which phenolic acids are the main compounds that attribute for most of the functional properties of many fruits. The aim of this study was to characterize the pomegranate extract towards development of phenolic acids based on a non-opioid substitution therapy for in vivo and in vitro studies. Pomegranate was extracted using two different solvents with three different methods of extraction. The quantification of phenolic acids was performed using High-Performance Liquid Chromatography (HPLC). The level of Mμ Opioid Receptor (MOR) and cyclic Adenosine Monophosphate (cAMP) was analysed by using an in vitro assay to determine the morphine dependence. Meanwhile, the morphine withdrawal effect was investigated through an in vivo study by analysing the memory impairment of rats using Morris Water Maze (MWM) and by determining the level of Brain Derived Neurotropic Family (BDNF) and cAMP response element binding protein (CREB). The result reveals that the pomegranate extract with water had the highest phenolic acid composition of gallic acid, ferulic acid, and ellagic acid. In the in vitro study, it shows that treatment of morphine with extract significantly reduced the level of cAMP and increased the level of MOR as compared to morphine and methadone treatments. While in the in vivo study, rats were divided into normal, morphine, morphine + methadone, and morphine + pomegranate groups. Data obtained from MWM show memory impairment of morphine group for time escape from pool was significantly higher than other groups of treatment. Meanwhile, BDNF and CREB levels in serum from morphine + pomegranate group were significantly higher than morphine and morphine + methadone groups. As conclusion, this study suggests that the phenolic acids in the pomegranate extract has a potential to be implemented as a non-substitution therapy for morphine-dependent treatment.

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