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

Mohd Adzim Khalili, Rohin and Atif Amin, Baig and Norhaslinda, Ridzwana and Mimie Noratiqah, Jumlia (2018) Characterization of a pomegranate (Punica granatum) extract towards development of anthocyanin based on a non-opioid substitution therapy in in vitro and in vivo studies. Health and the Environment Journal, 9 (1). pp. 14-27. ISSN 2180-1126

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Abstract

Pomegranate is one of the oldest tropical and subtropical edible fruits. Keeping in view of its rich in constituents and antioxidants, it has been studied scientifically and its uses have been verified. Currently, some compounds in pomegranate have been proven to induce cognitive improvements, related signaling in managing withdrawal symptoms. The present study investigated the characterization of anthocyanin from the pomegranate extract towards non-opioid substitution therapy in in vitro and in vivo studies. Anthocyanin was analyzed from pomegranate extract using high-performance liquid chromatography. The MORs and cAMP levels in the cells were determined using ELISA Kit from Cusabio Biotech and Cell Biolabs. The memory impairment in male Sprague-Dawley rats was determined using Morris water maze method. Then, the CREB and BDNF levels in serum rats were determined using ELISA Kit Abcam and Cell Signaling Technology. The water extract resulted in the highest contained of anthocyanin. The extract then was used for co-treatment for in vitro and in vivo studies. Co-treatment of morphine and pomegranate extract significantly reduced the production of cAMP level at lower concentration (p < 0.05). In memory impairment test, treatment group significantly reduced the distance and time latency compared to morphine group (p < 0.05). This is in correlation with levels of CREB and BDNF that were also significantly higher than morphine group (p < 0.05). These findings suggest that anthocyanin had potency as non-opioid substitution therapy that could reduce the dependence and withdrawal effects on chronic morphine treatment. As per knowledge, this is the first finding of using pomegranate extract in opioid treatment.

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