Phytochemical Screening, Evaluation of Antioxidant and Anti-Sickling Activities of Two Polar Extracts of Combretum Glutinosum Leaves
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British Journal of Applied Science & Technology 19(3): 1-11, 2017; Article no.BJAST.31563 ISSN: 2231-0843, NLM ID: 101664541 SCIENCEDOMAIN international www.sciencedomain.org Phytochemical Screening, Evaluation of Antioxidant and Anti-sickling Activities of Two Polar Extracts of Combretum glutinosum Leaves. Perr. ex DC Cheikh Sall 1*, Samba Fama Ndoye 2, Mbaye Diaw Dioum 2, Insa Seck 2, Rokhaya Sylla Gueye 2, Babacar Faye 3, Cheikhou Omar Thiam 2, Matar Seck 2, Papa Madièye Gueye 4, Djibril Fall 2, Mamadou Fall 5 3 and Tandakha Ndiaye Dieye 1Laboratory of Chemistry, Training and Research Unit of Health, Thies University, PB 967 Thies, Sénégal. 2Laboratory of Organic and Therapeutic Chemistry, Faculty of Medicine, Pharmacy and Odontology (F.M.P.O.), Cheikh Anta Diop University (U.C.A.D.), PB 5005 Dakar-Fann, Sénégal. 3Laboratory of Immunology, Pharmacy and Odontology (F.M.P.O.), Cheikh Anta Diop University (U.C.A.D.), Sénégal. 4Laboratory of Pharmaceutical Biochemistry, Pharmacy and Odontology (F.M.P.O.), Cheikh Anta Diop University (U.C.A.D.), Sénégal. 5Laboratory of Toxicology and Hydrology, Pharmacy and Odontology (F.M.P.O.), Cheikh Anta Diop University (U.C.A.D.), Sénégal. Authors’ contributions This work was carried out in collaboration between all authors. Author MS designed the study, performed the statistical analysis. Authors MS and CS wrote the protocol and wrote the first draft of the manuscript. Authors SFN, MDD, IS and RSG managed the analyses of the study. Authors BF, COT, PMG, DF, MF and TND managed the literature searches. All authors read and approved the final manuscript. Article Information DOI: 10.9734/BJAST/2017/31563 Editor(s): (1) Jesus F. Arteaga, Department of Chemical Engineering, Physical Chemistry and Organic Chemistry, University of Huelva, Spain. Reviewers: (1) Bouyahya Abdelhakim, Mohamed V University, Rabat, Morocco. (2) Viduranga Waisundara, Rajarata University of Sri Lanka, Mihintale, Sri Lanka. (3) Lai How Yee, Taylor’s University, Malaysia. Complete Peer review History: http://www.sciencedomain.org/review-history/18093 Received 13 th January 2017 Accepted 27 th February 2017 Original Research Article th Published 7 March 2017 _____________________________________________________________________________________________________ *Corresponding author: E-mail: [email protected], [email protected]; Sall et al.; BJAST, 19(3): 1-11, 2017; Article no.BJAST.31563 ABSTRACT Sickle cell disease is a major public health problem in Africa and many other areas across the world. In Senegal, a lot of plants are proposed by traditional healers to manage the sickle cell disease, among them Combretum glutinosum . Antisickling activity of Combretum glutinosum leaves extracts was studied in this work. Methanolic and ethyl acetate extracts of Combretum glutinosum leaves were evaluated on SS sickles type to determine their anti-sickling potential. Antiradical properties of methanolic extract were evaluated using the DPPH radical as oxidant. Total phenolic content of the methanol extract was determined. Phytochemical screening of the crude extract of methanol revealed the presence of tannins, saponins, phenols and flavonoids. Results show a good antisickling effect of extracts with a maximum antisickling revers of 81% and 69% respectively, of the methanol and ethyl acetate extracts at 10 mg/mL versus 67% for arginine at 120 min of incubation. The measured IC 50 were 0.65 and 0.163 for respectively the methanol extract and ascorbic acid. Antiradical powers 0.155 and 0.62 respectively for methanol extract and ascorbic acid were calculated from the effective concentrations. The results of this study confirm the traditional use of Combretum glutinosum leaves in the management of sickle cell disease. Keywords: Sickle cell disease; oxidative stress; medicinal plant; Combretum glutinosum; antisickling activity; antioxidant activity. 1. INTRODUCTION Indeed Sickle cell disease is a form of haemoglobinopathy that is characterized by Medicinal plants are used in the whole world and chronic hemolytic anemia. SCD is a genetic have a growing economic importance. The use of disease in which the SS individual possesses plant parts to treat human disease is as old as an abnormal ß-globin gene. A single base disease itself [1]. According to the World Health substitution in the gene encoding the human ß- Organisation (WHO), as many as 80% of world globin subunit results in the replacement of ß6 population, particularly in developing countries, glutamic acid by valine, which leads to the have used at least one time in their lives, devastating clinical manifestations of SCD. This traditional medicines for their primary health care substitution is followed by a remarkable need today and 25% of drugs are based on inflammatory response and oxidative stress plants and their derivatives [2]. Combretum within the blood circulation [7]. It also causes a glutinosum is a very large genus comprising drastic reduction in the solubility of sickle cell of about 250 species found in both temperate hemoglobin when deoxygenated, frequent chest and tropical regions in the world [3]. It is pain and vaso-occlusive complication [8]. SCD largely distributed in Africa throughout the occurs world-wide and the African continent Sahel belt from Senegal to Cameroon and bears the highest burden. Hispanic, Arabic, eastwards to the Sudan. It is a bushy shrub or Mediterranean and some Asian populations small tree growing up to 12 m and deciduous are also affected. According to Tim [9], species sprouting in the middle of the approximately 250,000 children are born each dry season. The plant parts are used in year with sick cell disease and over 70,000 traditional medicine in Africa to treat many individuals in the U.S. suffer from the disease. In human diseases. In Nigeria, the roots, the stem, Africa, SCD is a major public health problem with the bark and the leaves of Combretum a prevalence of SCD trait between 10 and 40% glutinosum are used in the treatment of scrotal [10]. In Senegal it is between 8 and 10% in the elephantiasis, dysentery, ring worms, bacterial general population [11]. Due to the high mortality diseases, syphilis, typhoid fever etc [4]. rate of sickle cell patients, especially in children, Antidiarrheal activity of aqueous young and since chemotherapy has its adverse effects, leaves extract of Combretum glutinosum there is need for rational drug development that was demonstrated [5]. Anti-malarial and anti- must embrace not only synthetic drugs but also schistosomal activities of Combretum glutinosum natural products. It is acknowledged world-wide were respectively described by Yahaya and that traditional medicine can be explored and Ouattara [2,6]. Therapeutic potential of different exploited for enhanced health management. The parts of Combretum glutinosum remains to be ever increasing demand for safer and cheaper explored on several other diseases such as herbal recipes in developing country, has led to sickle cell disease (SCD). the extraction and development of several drugs 2 Sall et al.; BJAST, 19(3): 1-11, 2017; Article no.BJAST.31563 and chemotherapeutic agent from plants, as well The age of patients ranged from 18 to 35 years as from traditional herbal remedies [5] in both sexes. Sampling is made at the elbow crease of the patients and the blood was In recent years, the interest in natural collected in EDTA tubes. Chemical and antioxidants, in relation to their therapeutic biochemical products used in this work were properties, has increased dramatically. Scientific obtained from different suppliers (Prolabo, research in various specialties has been Scharlau, Aldrich or Carlo Erba). All solvents developed for extraction, identification and were freshly distilled before use. The human quantification of these compounds from several ethics approve was obtained from Cheikh Anta natural substances namely, medicinal plants and Diop University of Dakar under the number food products [12,13]. The antioxidant activity of 0228/2017/CER/UCAD on February 03 2017. a compound corresponds to its ability to resist oxidation. The best known antioxidants are the β- 2.1.2 Plant material carotene (provitamin A), ascorbic acid (vitamin C), tocopherol (vitamin E) as well as phenolic The leaves of Combretum glutinosum used in compounds. Indeed, most synthetic or naturally this work were purchased from traditional healers occurring antioxidants have phenolic hydroxyl in Tilene market in Dakar. The identification was groups in their structures; and antioxidant carried out at the Laboratory of Pharmacognosy properties are attributed in part to the ability of of the Medicine, Pharmacy and Odontology these natural compounds to scavenge free Faculty at Cheikh Anta Diop University of Dakar. radicals such as hydroxyl radicals (OH •) and The dried leaves were crushed and powdered superoxide (O 2• ) [14,15]. Several methods are using a Brabender mill. used to evaluate, in vitro and in vivo antioxidant activity by scavenging different radicals such as 2.2 Methods • peroxyl radical (ROO ) by ORAC and TRAP methods [16]; ferric ions by FRAP [17]; or the 2.2.1 Extraction of the Combretum ABTS • radical [18] and the method using free glutinosum leaves powder radical DPPH• [19]. Because it takes place at room temperature and can eliminate the risk of Leaves powder 60 g of Combretum glutinosum thermal degradation of thermolabile molecules, were extracted with 300 mL of ethyl acetate. The the DPPH• radical test is the most recommended mixture was stirred