Phytoconstituents and Bioactivities of the Bark of Pleiogynium Timorense (DC.) Leenh (Anacardiaceae)
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J Herbmed Pharmacol. 2020; 9(1): 20-27. http://www.herbmedpharmacol.com doi: 10.15171/jhp.2020.03 Journal of Herbmed Pharmacology Phytoconstituents and bioactivities of the bark of Pleiogynium timorense (DC.) Leenh (Anacardiaceae) Gehan Fawzy Abdel Raoof 1* ID , Ataa Abdelhaleem Said1, Khaled Younes Mohamed2, Hesham A. Gomaa3,4 1Pharmacognosy Department, National Research Centre, Dokki, Giza, Egypt 2Internal Medicine Department, Medical Division, National Research Centre, Giza, Egypt 3Biochemistry Department,Faculty of pharmacy, Nahda university ,Beni-Suef, Egypt 4Pharmacology Department, College of Pharmacy, Jouf University, Sakakah, Saudi Arabia A R T I C L E I N F O A B S T R A C T Article Type: Introduction: The purpose of this study was to evaluate the phytoconstituents and various Original Article bioactivities of Pleiogynium timorense bark as a step towards the production of a new drug from natural origin to overcome the complications of the synthetic drugs. Article History: Methods: The phenolic compounds were isolated and identified by chromatographic and Received: 13 February 2019 spectroscopic methods as ultra violet (UV) and nuclear magnetic resonance (NMR) spectra. Accepted: 2 June 2019 The isolated compounds, as well as 70% methanol extract of P. timorense bark were tested for cytotoxicity against human colon carcinoma (HCT 116), human hepatocellular liver carcinoma Keywords: (HepG2), normal melanocytes (HFB-4) and human breast carcinoma (MCF-7) cell lines. In Antihyperglycaemic agents addition, the methanol extract was evaluated for renal protective, hepatoprotective, antioxidant Antioxidant and antihyperglycaemic activities. Cytotoxic activity Results: Seven phenolic compounds were isolated from the bark of the plant for the first time Hepatorenal toxicity which were identified as; pyrogallol, catechin, gallic acid, kaempferol, quercetin, rutin and Phenolic compound quercetrin. Moreover, the methanol extract of the bark showed a promising cytotoxic effect Pleiogynium against HepG2 cell line more than that of the isolated compounds comparing with doxorubicin (a positive control), where catechin and gallic acid showed moderate effects. In addition, the methanol extract showed potent antioxidant, hepatorenal protective and antihyperglycaemic effects. Conclusion: Pleiogynium timorense extract possesses a potent cytotoxic effect against HepG2 cell line and significant antioxidant, hepatorenal protective and antihyperglycaemic effects. Implication for health policy/practice/research/medical education: The results of this study suggest that the methanol extract of Pleiogynium timorense bark, as well as catechin and gallic acid can be used as promising cytotoxic agents against HepG2 cell line. Moreover, the methanol extract can be used as a potent antioxidant, hepatorenal protective and antihyperglycaemic agent upon further clinical studies. Please cite this paper as: Abdel Raoof GF, Said AA, Mohamed KY, Gomaa HA. Phytoconstituents and bioactivities of the bark of Pleiogynium timorense (DC.) Leenh (Anacardiaceae). J Herbmed Pharmacol. 2020;9(1):20-27. doi: 10.15171/jhp.2020.03. Introduction against Staphylococcus aureus and Bacillus subtilis (5). The production of drugs from plant origin aims to avoid Another study showed that the alcoholic extract of the leaves the complications of synthetic origin agents. Pleiogynium had antioxidant, anti-inflammatory and hypoglycemic timorense (DC.) Leenh. (Anacardiaceae) is native to effects. Moreover, myricetin-3-O-α-L-rhamnopyranosid, Australia, with the common name of Gambozia and is kaempferol, kaempferol-3-O-β-D-glucopyranoside, described as an ornamental tree (1,2). Anacardiaceae quercetin -3-O-β-D-glucopyranoside 1,3,4,6-tetra-O- contains many plants with edible seeds and fruits (e.g., galloyl-β-D-glucopyranose, 1,4,6-tri-O-galloyl-β-D- Cashew nuts, Pistachio nuts and mango (3). The fruits of glucopyranose, 3,5-di-O-galloylquinic acid,quercetin- P. timorense (DC.) Leenh. can be eaten and used in making 3-O-β-D-galactopyranoside, gallic acid, kaempferol-3- jams and jellies (4). Rutin, myricetin, hyperin, quercitrin, O-β-D-6”-methyl glucuronopyranoside, kaempferol- quercetin, β-sitosterol and lupeol have been isolated from 3-O-β-D-galactopyranoside and kaempferol-3-O-β-D- the leaves which possessed potent antimicrobial effects glucuronopyranoside were isolated and identified from *Corresponding author: Gehan Fawzy Abdel Raoof, Email: [email protected] Phytoconstituents and bioactivities of Pleiogynium timorense Bark the leaves of the plant (6). Cyanidin-3-glucoside was described (13,14). isolated from Pleiogynium fruits which showed a potent antioxidant activity (7). In our previous work, rutin, HPLC determination of phenolics and flavonoids catechin, quercetrin and quercetin were isolated from the The identification and quantification of flavonoids and pericarp of the plant, as well as the methanol extracts of phenolics in 70% methanol extract of P. timorense bark the pericarp and seeds and showed anti-inflammatory, were performed by HPLC according to the previously antioxidant, analgesic, hepatorenal protective effects (8). described methods (15,16). Moreover, GC/MS analysis of the lipoidal matter of the seeds showed that 1-heptene was the major compound in Extraction and isolation unsaponifiable matter, while linoleic acid was the major The air dried powdered bark of P. timorense (1 kg) was fatty acid (9). Furthermore, dichloromethane extract of extracted with petroleum ether (60-80°C) for defatting. the bark showed a significant activity against the A2780 The defatted powder was extracted with 70% methanol human ovarian cancer cell line due to the presence of three by percolation, then the concentrated extract (70 g) was new trihydroxy alkylcyclohexenones which were isolated phytochemically screened using the standard procedures from this extract (10). In our recent work the constituents which were previously described (13) to identify its of P. timorense pericarp and seeds were identified by constituents. The extract (40 g) was subjected to polyamide using high-performance liquid chromatography (HPLC) CC and eluted with gradient water: methanol (10:0) to with electrospray ionization mass spectrometry (11). (0:10) to give five fractions. Fraction 1, one compound Moreover, the volatile constituents of P. timorense fruits which was eluted with water: methanol (90:10), was were identified which showed a moderate cytotoxic effect purified on Sephadex LH-20 CC and was eluted with on human hepatoma cells and a powerful cytotoxic effect methanol to give compound 1. Fraction 2, two compounds against laryngeal carcinoma and breast adenocarcinoma which were eluted with water: methanol (80:20), was human tumor cell lines (12). The present study aims to the applied on Sephadex LH-20 C using methanol as eluent methanol extract of the bark of this plant as antioxidant, to give compounds 2 and 3. Fraction 3, two compounds antihyperglycaemic, cytotoxic and hepatorenal protective which were eluted with water: methanol (70:30), was agent with the aim of producing a natural drug. applied on Sephadex LH-20 C to give compounds 4 and 5. Fraction 4, one compound which was eluted with water: Materials and Methods methanol (60:40), was purified on Sephadex LH-20 C Equipments, materials and chemicals to give compound 6. Fraction 5, one compound which Solvent mixtures; S1: BAW (n-butanol: acetic acid: water was eluted with water: methanol (50:50), was purified on (4:1:5, v/v/v) upper phase), S2: (water: glacial acetic acid Sephadex LH-20 C to give compound 7. (85:15, v/v)) and S3: (methanol: chloroform (30:70)). Thin layer chromatography (TLC) was done on TLC silica gel Cytotoxicity assay procedures F254 plates (200 mm layer thickness) (Merck; Darmstadt, Human tumor cell lines Germany), column chromatography was done on Silica Human breast carcinoma (MCF-7), normal melanocytes gel (0.063–0.200 mm) and Sephadex LH-20 (Pharmacia (HFB-4), human hepatocellular liver carcinoma (HepG2) Fine Chemicals). Nuclear magnetic resonance (NMR) and human colon carcinoma (HCT116) cell lines were spectra were measured on JEOL EX-500 spectrometer obtained from the American Type Culture Collection and (Tokyo, Japan) with 500 MHz for 1H-NMR and 125 MHz were maintained by serial sub-culturing in the National for 13C-NMR. MS was done using Finnigan MAT SSQ Cancer Institute, Cairo, Egypt. 7000, 70 eV. Ultraviolet spectra were recorded by using UV/VIS: Shimadzu UV, chromatograms were visualized Culture media under visible recording spectrophotometer model-UV The cells were suspended in RPMI 1640 medium (SIGMA 240 (NRC, Egypt). ALORICH) supplemented with 10% fetal calf serum (SIGMA, USA) in presence to 1% antibiotic antimycotic Plant identification and collection mixture (10.000 U/mL K-penicillin, 10.000 μg/mL The bark of P. timorense was prepared from Zoo garden, streptomycin sulphate and 25 μg/mL amphotericin B) and Giza, Egypt. The plant was identified by Dr. M. El-Gebaly, 1% L-glutamine (all purchased from Lonza, Belgium). the taxonomist at the Department of Botany, National Research Centre (NRC), Giza, Egypt. A voucher specimen Assay method for cytotoxic activity (possessing number 2001) was kept in NRC. The cytotoxicity against HCT116, Hep-G2, HFB4 and MCF-7 cells were tested in the National Cancer Institute, Phytochemical screening according to the SRB (Sulforhodamine B) assay using MTT The constituents of the methanolic extract of the bark (3-(4,5-dimethylthiazol2-yl)- 2,5-diphenyltetrazolium were identified