Abstract

 This study was conducted with the aim to extract and purify a polyphenolic compound “ Resveratrol†from the skin of black grapes Vitis vinifera cultivated in Iraq. The purified resveratrol is obtained after ethanolic extraction with 80% v/v solution for fresh grape skin, followed by acid hydrolysis with 10% HCl solution then the aglycon moiety was taken with organic solvent
 ( chloroform). Using silica gel G60 packed glass column chromatography with mobile phase benzene: methanol: acetic acid 20:4:1 a partial purified resveratrol was obtained then preperative thin layer chromatography technique yielded pure crystals identified as resveratrol (mixture of two isomers cis and trans) in relation to resveratrol standard (35 mg resveratrol crystals / 0.5 kg fresh grape skin was obtained as a result of these processes ). The study was also employed an in vitro evaluation the cytotoxic effect of pure resveratrol on some cell line including : the murine mammary adenocarcinoma AMN-3 cell line, the human laryngeal carcinoma (Hep-2) cell line and the Rat embryo fibroblast (Ref) cell line, at different concentrations and different expousure time. The cytotoxic effect of the pure resveratrol was studied in comparison with trans- resveratrol standard in concentrations of (12.5, 25, 50 and 100) µg/ml for both purified resveratrol and the standard, also the comparism included methotrexate drug in concentrations (0.05 , 0.1 , 0.2 and 0.4) µg/ml toward the growth effects of the three types of cell lines and at three exposure times (24, 48 and 72) hours. The cytotoxic inhibition effect for the purified extracted resveratrol revealed that the highest significant effect (P<0.01) was achieved after 24 hr of exposure on both AMN-3 and Ref cell lines. Hep-2 cell line responded to extracted resveratrol in different manners.
 Keywords: Resveratrol , grape skin , polyphenols, cytotoxic study
Highlights
Cancer is a complex set of more than 200 diseases with many causes and multiple stages and histological grades of malignancy [1].result of exogenous environmental, lifestyle and host genetic factor which attributed about 2% of cancer factors [2].Cancer treatments including surgical, radiotherapy, chemotherapy, and biotherapy by immunization and gene therapy, are employed as conventional treatments which may not always be satisfactory to relieve sever symptoms or even cure cancer with untolerated side effects and many get resistance [3]
Three cell lines were studied (AMN-3, Hep-2 and Rat embryo fibroblast (Ref) cell line) at three exposure time 24,48 and 72 hours using two fold dilutions to get concentration ranging from 12.5 μg/ml to 100 μg/ml for both the purified extracted resveratrol and the trans- resveratrol standard, while for methotrexate drug the concentrations included (0.05, 0.1, 0.2 and 0.4) μg/ml according to Al-Shemary study [24]
The cytotoxic effect of the purified extracted resveratrol on Hep-2 cell line was shown in table(2)
Summary
Cancer is a complex set of more than 200 diseases with many causes and multiple stages and histological grades of malignancy [1].result of exogenous environmental, lifestyle and host genetic factor which attributed about 2% of cancer factors [2].Cancer treatments including surgical, radiotherapy , chemotherapy, and biotherapy by immunization and gene therapy, are employed as conventional treatments which may not always be satisfactory to relieve sever symptoms or even cure cancer with untolerated side effects and many get resistance [3]. The search for novel and effective cancer chemopreventive agent has led to the identification of various naturally occurring phytocompounds, one of which is resveratrol (trans- 3,4′ ,5 – trihydroxy stilbene) a phytoalexin drived from grape skin and other fruits [6]. As there is drug development; programmes for pre- clinical screening of the vast numbers of chemicals for specific and non- specific cytotoxicity against many types of cells are involved. Both are important for indicating potential therapeutic target and safety evaluation[13]. The secondary metabolite “ Resveratrol” was synthesized by the plant in response to stress , including disease and ultraviolet light , as phytoalexin was first reported in skins of grapes by Creasy and Coffee[15] ,in 1988 later in 1991 was reported by Siemann and Creasy[16] , from wines
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Schedule a callMore From: Iraqi Journal of Pharmaceutical Sciences ( P-ISSN: 1683 - 3597 , E-ISSN : 2521 - 3512)
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