Antibacterial, Antioxidant and Cell Proliferative Properties of Coccinia Grandis Fruits Prashant Sakharkar1, Balwantsinh Chauhan2,*

Original Research Article Antibacterial, antioxidant and cell proliferative properties of Coccinia grandis fruits Prashant Sakharkar1, Balwantsinh Chauhan2,* 1Department of Clinical & Administrative Sciences, Roosevelt University College of Pharmacy, 1400 N. Roosevelt Blvd., Schaumburg, IL, 60173, USA 2Department of Bipoharmaceutical Sciences, Roosevelt University College of Pharmacy, 1400 N. Roosevelt Blvd., Schaumburg, IL, 60173, USA Article history: Abstract Received: Jun22, 2016 Objective: Little knowledge is available on the antimicrobial and Received in revised form: Jan 16, 2017 antioxidant properties of Coccina grandis fruits and no study has Accepted: Jan 22, 2017 reported on its cell proliferative property. The aim of this study Vol. 7, No. 4, Jul-Aug 2017, was to examine the antimicrobial, antioxidant and cell proliferative 295-307. property of fruits of C. grandis. Materials and Methods: Fruits of C. grandis were extracted * Corresponding Author: using water; ethanol and acetone by cold and hot Soxhlet Tel: (+1) 847-330-4532 extraction. The antibacterial activities of the extracts were tested Fax:(+1) 847-330-4525 against Staphylococcus aureus, Enterococcus faecalis, Escherichia ‎[email protected] coli and Pseudomonas aeruginosa using the modified Kirby-Bauer diffusion method and compared against erythromycin. The Keywords: Coccinia grandis antioxidant property was determined using Cayman's antioxidant Antibacterial assay; whereas cell proliferation/cytotoxic properties were Antioxidant evaluated using the Cell Titer 96 Aqueous One Solution Cell MTS Cell proliferation assay with MDA-MB 321 breast cancer cells. Data were analyzed Ivy gourd for correlation and differences using unpaired student's t-test and one-way ANOVA. A p value of <0.05 was considered statistically significant. Results: Both cold and hot ethanol and acetone extracts of C. grandis fruits showed some degree of bacterial growth inhibition. Acetone extracts exhibited higher antibacterial activity. Both ethanol extracts showed antioxidant property when compared with standard Trolox. In contrary to cytotoxicity, all four extracts showed cell proliferation compared to controls at different concentrations. However, acetone extracts exhibited greater cell proliferation compared to ethanol extracts and cold extracts performed better than the hot extracts. Conclusion: C. grandis fruits exhibited some degree of antimicrobial, antioxidant and cell proliferative properties. Further investigation is warranted to isolate, confirm and characterize phytochemicals that are responsible for the medicinal properties observed. Please cite this paper as: Sakharkar P, Chauhan B. Antibacterial, antioxidant and cell proliferative properties of Coccinia grandis fruits. Avicenna J Phytomed, 2017; 7 (4): 295-307. AJP, Vol. 7, No. 4, Jul-Aug 2017 295 Sakharkar et al. Introduction plant has been considered of having some The concept of combining dietary medicinal value (Yadav et al., 2010; constituents to manage various illnesses is Nagare et al., 2015). Plant preparations historically part of most of the cultures. from C. grandis are indigenously used for Currently, there is wide spread use of various skin diseases, bronchitis, anorexia, complementary and alternative medicine cough, asthma, catarrh, and epilepsy. (CAM) globally. Of five categories of Moreover, in Unani systems of medicine it CAM identified, the herbal products are has been used for ringworm, psoriasis, the most abundantly used form of the small pox and scabies (Kirtikar and Basu, therapy. The search for new herbal 1994; Philcox, 1997; Nagare et al., 2015). products/drugs for different human Plant preparations are also used for itchy ailments is increasing, as they are believed skin eruptions, wound healing, leprosy, to be less or non-toxic in nature. It is well gonorrhea, pyelitis, cystitis, snakebite, recorded that various plants belonging to malarial infection, infective hepatitis, and Cucurbitaceae family are used as herbal jaundice; Also, it has been given as a medicine in most of the culture. Coccinia hepatoprotective remedy and for treating plants (Ivy) belonging to Cucurbitaceae renal calculi (Kirtikar and Basu, 1994; family have their own importance in Vadivu et al., 2008; Shaheen et al., 2009; traditional medicines, including Ayurveda, Deshpande et al., 2011a; Dnyaneshwar and practiced in India, Chinese herbal Patil, 2011; Ramakrishnan et al., 2011; medicines practiced in China and Unani Sood et al., 2012). C. grandis is also system of medicine or Greco-Arab known for its anti-diabetic, anti-obesity, medicine practiced in Iran (Khan et al., antimicrobial, antifungal, antileishmanic, 1979). antioxidant, antihypertensive, antitussive, Ivy gourd is known in India by various antiulcer, analgesic, antipyretic, vernacular names like tondi in Marathi, antianaphylactic, and anti-cancer Tindora, Tinda and Kundu in hindi, properties (Kirtikar and Basu,1994; Yadav dondakaya in telugu, tomdekayi in et al., 2010; Tamilselvan et al., 2011; kannada, etc. (Ali et al., 2005). In other Pekamwar et al., 2013; Gill et al., 2014; parts of the world, it is known as Hong gua Nagare et al., 2015). However, its in Chinese, Bat in Vietnam, Pepasan in therapeutic efficacy is yet not conclusive Malay, Yasai, Karasuuri in Japanese, due to the lack of carefully controlled Gourds ecarlate de l'Inde in France, pepino scientific investigations. Although, most of Cimarron in Spanish and Skariagenagurk. the studies have either used extract of Ivy gourd (Coccinia grandis) is found in stem, root and most often leaf alone or in tropical Asia (India, Pakistan, Bangladesh, combination, only few studies tested Sri Lanka, Indonesia, Malaysia, the medicinal properties of C. grandis fruits Philippines, and Thailand), and Africa (Vadivu et al., 2008; Shaheen et al., 2009; (Cooke, 1903). Coccinia indica Wright Deshpande et al., 2011a; Dnyaneshwar and and Arn., and Coccinia cordifolia (L.) Patil, 2011; Ramakrishnan et al., 2011; Cogn., Cephalandra indica, Naud., and Sood et al., 2012). To our knowledge, Bryonia cordifolia (L.) Voigt.( Kirtikar there are hardly any in-vitro studies testing and Basu, 1994; Philcox,1997; Nagare et cell proliferative property of C. grandis al., 2015) are the other names of C. fruit. The goal of this study was to conduct grandis which is a climber, trailer, preliminary phytochemical screening and dioecious, and perennial plant. Young, to evaluate the potential antimicrobial, tender and long slender stem tops, leaves, antioxidant and cell proliferative property and tuberous roots of C. grandis are of C. grandis fruit. cooked or used as a seasoning and young fruits are used in salads. Every part of this AJP, Vol. 7, No. 4, Jul-Aug 2017 296 Cell proliferative properties of Coccinia grandis Materials and Methods (potassium mercuric iodide). Formation of Plant extract yellow creamy precipitates indicated the Un-ripened fresh fruits of C. grandis presence of alkaloids. were collected in June 2014 from a local c. Wagner’s reagent: Filtrate was Indian grocery store in Schaumburg, IL treated with 4-6 drops of Wagner’s reagent (USA) and authenticated at Department of (iodine in potassium iodide). Formation of Biology, Chicago State University in brown/reddish brown precipitation or Chicago, IL. Fresh fruits were washed with coloration indicated the presence of distilled water to clean any debris. Fruits alkaloids. were cut into small pieces and dried at room temperature away from direct Carbohydrates (Molisch’s test) sunlight. Dried material was weighed and Few drops of Molisch’s reagent were subjected to cold extraction using ethanol added to 2 mL of extract. This was (94-96 %, BDH) and acetone at 4oC, for 48 followed by gradual addition of 2 mL of hr with occasional stirring. Similarly, hot concentrated sulfuric acid down the side of extraction was carried out using a Soxhlet the test tube. The mixture was then apparatus (58oC; 7 cycles) with alcohol allowed to stand for two to three minutes. and acetone. Residues obtained on Formation of red or dull violet color at the extractions were evaporated to dryness interface of the two layers indicated the using ‘Rotavap’ and dried. All extracts presence of carbohydrates. were stored at 4oC until further use. Flavonoids Phytochemical screening: One mL of 10% lead acetate solution Dried fruit powder was extracted (at was added to 1 mL of the aqueous extract. 4oC) for 48 hr in MilliQ water and The formation of a yellow precipitate obtained residue was dissolved in small indicated the presence of flavonoids. volume of MilliQ water and labeled as “aqueous extract”. This aqueous extract Cardiac glycosides (Keller-Killiani test) along with ethanol and acetone extracts Crude extract (2-5 ml) was mixed with were subjected to preliminary 2 ml of glacial acid containing 1-2 drops of phytochemical screening using following 2% solution of ferric chloride. The mixture tests (Egwaikhide and Gimba, 2007; was then poured into another test tube Roopashree et al., 2008; Abba et al., 2009; containing 2 ml of concentrated sulfuric Njoku and Obi, 2009; Yadav and acid. A brown ring of a deoxy-sugar, Agarwala, 2011; Sood et al., 2012; characteristic of alcoholic cardenolides at Soloman et al., 2013). the interface indicated the presence of cardiac glycosides. Alkaloids Nine drops of 1% diluted hydrochloric Tannins (Braymer’s Test) acid was added to 6mL of extracts, mixed To 5ml of the extract, few drops of well and the mixture was left for some 0.1% ferric chloride were added. A time and filtered.

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