Hepatoprotective, Antihyperglycemic and Cytotoxic Activities Of
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Arom & at al ic in P l ic a n d t e s M Mostafa et al., Med Aromat Plants (Los Angles) 2017, 6:4 Medicinal & Aromatic Plants DOI: 10.4172/2167-0412.1000297 ISSN: 2167-0412 Research Article Open Access Hepatoprotective, Antihyperglycemic and Cytotoxic Activities of Jacaranda acutifolia Leaf Extract Nada M Mostafa1, Omayma A Eldahshan1, Hesham A El-Beshbishy2 and Abdel Nasser B Singab1* 1Department of Pharmacognosy, Ain Shams University, Abbassia, Cairo, Egypt 2Department of Medical Laboratories Technology, Faculty of Applied Medical Sciences, Taibah University, Al-Madinah Al-Munwarah, Saudi Arabia *Corresponding author: Abdel Nasser B Singab, Department of Pharmacognosy, Ain Shams University, Cairo, Egypt, Tel: 01005036231; E-mail: [email protected] Received date: June 30, 2017; Accepted date: July 13, 2017; Published date: July 20, 2017 Copyright: © 2017 Mostafa NM, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract Objective: Leaves methanol extract of Jacaranda acutifolia Humb. and Bonpl. (JA) family Bignoniaceae was subjected to phytochemical investigation as well as antioxidant, hepatoprotective, cytotoxic and antihyperglycemic activities evaluation. Key findings: Eight compounds were identified: luteolin-7-O-β-D-glucuronide, luteolin-7-O-β-D-glucoside, aesculetin, luteolin, verbascoside, luteolin-7-O-β-D-glucuronide methyl ester, apigenin-7-O-β-D-glucuronide methyl ester and apigenin. JA revealed a potent antioxidant activity in vitro superior to vitamin E (DPPH assay; EC50 of 0.43 mg/mL). A potential cytotoxic activity was produced against hepatocellular (HepG2) and cervical (HeLa) carcinoma cells with IC50 of 6.05 and 16.7 µg, respectively. Treatment with JA extract inhibited the rise in alanine aminotransferase and aspartate aminotransferase by 33.6% and 36.8% respectively, reduced thiobarbituric acid by 35.7% and decreased the tamoxifen-induced elevation in tumor necrosis factor alpha (TNF-α) level by 42.86%. JA extract elicited a significant decrease in fasting blood glucose by -59.26%. Conclusions: Jacaranda acutifolia could be a natural source for antioxidant, hepatoprotective supplements and could provide a basis for a potential cytotoxic agent. The compounds isolated are responsible at least in part for the observed effects. Keywords: Jacaranda acutifolia; Hepatoprotective; but with other compounds as homogentisic acid p-benzoquinone, Antihyperglycemic; Cytotoxic phenyl acetic acid and resorcinol. The oil showed activity against Staphylococcus aureus, Escherichia coli, Candida albicans, Salmonella Introduction typhimurium and Shigella flexneri where n-hexane extract showed moderate activities against many microorganisms [8]. The total Genus Jacaranda distributes around the world and includes 49 phenolic contents of Jacaranda acutifolia were calculated as 17.20 mg/g species. The main identified constituents are flavonoids, triterpenes, gallic acid equivalents. The ethyl acetate fraction possessed the highest quinones, and acetosides [1]. Members of this genus are well known in antioxidant activities, estimated by DPPH radical scavenging traditional ethnobotany for their promising pharmacological activities (EC50=0.049 mg/mL) and ferric ion reducing activities (EC50=0.125 [2]. mg/mL) [9]. Jacaranda species were used, traditionally, to treat rheumatism, The present study was undertaken to evaluate antioxidant, cytotoxic, leishmaniasis as well as venereal infections and gastrointestinal hepatoprotective and antihyperglycemic activities Jacaranda acutifolia disorders. Jacaranda acutifolia have shown diuretic and astringent leaves and to identify compounds responsible for the observed effects. activities [3]. Leaves and barks could be applied directly to the wounds or in the form of decoction or infusion, as they are considered as Materials and Methods disinfectant [4]. The bark could be used as astringent and diuretic and in the treatment dermatitis, syphilis and diseases related to urinary tract diseases [5]. Plant materials Jacaranda acutifolia leaves contain verbascoside, jacaranone, phenyl Jacaranda acutifolia (Humb. and Bonpl.) leaves (JA), Bignoniaceae acetic-β glucoside, scutellarein-7-glucoronide and hydroquinone [6]. were collected from Merryland Botanical Garden, Cairo, Egypt, and Flowers contain anthocyanins which are responsible for their violet air-dried. They were authenticated by Prof. Abd El Salam Mohamed color [5]. A novel biflavonoid [kaempferol (6→8″) apigenin] was Al-Nowiahi, Professor of Taxonomy, Faculty of Science, Ain Shams isolated from the leaves [7]. It exerted a promising cytotoxic activity University. Voucher specimens of JA (PHG-P-JA-201) was deposited at against breast carcinoma cell line (MCF-7). The main components of Department of Pharmacognosy, Faculty of Pharmacy, Ain Shams leaf volatile oil of Jacaranda were methyl linolenate (26.7%), 1-octen-3- University, Cairo, Egypt. ol (10.8%), methyl phenyl acetate (9.9%), beta-linalool (5.5%) and palmitic acid (4.7%). n-Hexane extract revealed similar composition, Med Aromat Plants (Los Angles), an open access journal Volume 6 • Issue 4 • 1000297 ISSN: 2167-0412 Citation: Mostafa NM, Eldahshan OA, El-Beshbishy HA, Singab AN (2017) Hepatoprotective, Antihyperglycemic and Cytotoxic Activities of Jacaranda acutifolia Leaf Extract. Med Aromat Plants (Los Angles) 6: 297. doi:10.4172/2167-0412.1000297 Page 2 of 8 Processing of Jacaranda acutifolia leaves saline for seven successive days. Group II: Rats were injected daily (i.p.) with TAM for seven successive days at a dose of 45 mg/kg/day in The intact air-dried plant materials (2 Kgs) were boiled in distilled normal saline. Group III: Rats were treated daily i.p. with JA extract at water for 2 hours then filtered while hot. The water extract was a dose of 20 mg/kg/day in normal saline for 7 successive days then i.p. lyophilized, and then extracted with methanol. The methanol portions injection of TAM at a dose of 45 mg/kg/day in normal saline for 4 were distilled off in rotary evaporator at 55˚C till dryness. The extract successive days starting at day 4-7. was concentrated until constant weight (87 g) then kept in vacuum desiccators over anhydrous CaCl2. Dosing volume was kept at 10 mL/kg. After the last treatment by 24 h, animals were subjected to light ether anesthesia and blood samples Experimental animals were collected by orbital puncture in serum-separating tubes. The blood was centrifuged at 3000 rpm for 10 min to separate the serum. Female Sprague-Dawley rats (120-140 g) were used. Animals were Sera were used for measurement of ALT and AST activities. maintained under standard conditions of temperature (24 ± 5˚C) and relative humidity (55 ± 5%), with a regular 12 h light: 12 h dark cycle, Anaesthetized animals were scarified by cervical dislocation. The and allowed free access to standard laboratory food and water 7 days liver was rapidly dissected out, washed in ice-cold isotonic saline and before starting the experiment and during the whole period of the blotted between two filter papers. Part of each liver was used to prepare experiment. 10% (w/v) liver homogenates in ice-cold 0.1 M potassium phosphate buffer, pH 7.5, and stored at -70 ˚C for subsequent analysis. All animals were fed with common pellet diets and water ad libitum. All stressful conditions were avoided. All animals were treated Antihyperglycemic activity humanely in accordance with the guidelines for animal’s care set by the World Health Organization. Experimental protocol was approved by Induction of diabetes mellitus: Diabetes was induced in the rats by a the local committee for animal care, Taibah University, Al-Madinah single (i.p.) injection of freshly prepared STZ (60 mg/kg b.w.) in Al-Munwarah, Saudi Arabia. normal saline. Two days after STZ administration, blood samples were obtained from the tips of the rat’s tail and the fasting blood glucose Fraction II (24.8 g) was re fractionated over a cellulose column and (FBG) levels determined using OneTouch® Ultra® glucometer (Life eluted with distilled water followed by 50% methanol in water, then Scan, USA) to confirm diabetes. The diabetic rats exhibiting blood neat methanol. Sub-fraction A yielded compounds 1, 2 and 3, while glucose levels above 190 mg% were included in this study [12]. The sub-fraction B yielded compound 4. Compounds were further purified biochemical effects of the extracts were compared to GLB. by preparative PC. Experimental animals' treatment: In the present study animals were Fraction III (26.9 g) was re-fractionated over a polyamide column. divided into four groups (eight rats in each): Group I: Normal control Elution started with distilled water then gradually decreasing polarity rats received 0.1 mL DMSO and 0.5 mL 5% Tween 80 for 7 days by oral by addition of methanol in water with different proportions, followed lavage. Group II: STZ-diabetic rats received 0.1 mL DMSO and 0.5 mL by neat methanol. Sub-fractions were pooled together to give five main 5% Tween 80 for 7 days by oral lavage. Group III: Diabetic rats orally subfractions: A-E. Sub-fraction B (30% MeOH in water) yielded received GLB (20 mg/kg/day) in 0.1 mL DMSO and 0.5 mL 5% Tween compound 5 by preparative PC on Whatmann No. 3 MM and 6% 80 for 7 days by oral lavage [13]. Group IV: STZ-diabetic rats orally acetic acid as solvent system, then purified on a column packed with received JA extract dissolved in 0.1 mL DMSO and 0.5 mL 5% Tween sephadex LH-20. Sub-fraction C (50% MeOH in water) yielded two 80 at a dose of 20 mg/kg/day for 7 days by oral lavage [14]. On the 8th compounds 6 and 7 which were isolated by preparative PC using BAW day, the fasting rats were subjected to light ether anesthesia and killed and 6% acetic acid as solvent systems individually. by cervical dislocation. Trunk blood was collected into heparinized Fraction IV eluted with 75% methanol in water (8.3 g) was re- chilled tubes containing NaF.