Vol 9, Issue 6, 2016 Online - 2455-3891 Print - 0974-2441 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 IN VITRO ANTI-SICKLING ACTIVITY OF ARTEMISIA HERBA-ALBA ASSO METHANOLIC EXTRACT ON SICKLE CELL DISEASE NESSRIN GHAZI ALABDALLAT* Department of Medical Laboratory Sciences, Collage of Applied Medical Sciences, Majmaah University, Majmaah, Saudi Arabia. Email: n.alabdallat@mu.edu.sa Received: 27 April 2016, Revised and Accepted: 10 May 2016 ABSTRACT Background: Sickle cell disease (SCD) is caused by polymerization of abnormal hemoglobin S when oxygen tension decreases. Previous studies have been indicated that some medicinal plants have shown an anti-sickling activity, which indicates a new therapeutic way to a range of people who are affected by this hemoglobinopathy. The current study aimed to assess the in vitro anti-sickling activity of Artemisia herba-alba Asso methanolic extract. Methods: The blood samples used in the evaluation of the anti-sickling activity of the plant extract in this study were taken from patients known to have SCD, attending in the King Khaled Hospital in Majmaah. Emmel test was used to assess anti-sickling activity of this plant. Results: The normal shape of the red blood cells (RBCs) was observed after incubation of RBCs with A. herba-alba Asso extract and 2% sodium metabisulfite as compared to control. A significant increase in the percentage of unsickled RBCs was observed after incubation of RBCs with 2% sodium metabisulfite in the presence of 500 and 1000 µg/ml of A. herba-alba Asso extract. Besides, the difference between the percentage of unsickled RBCs after 30 and 60 minutes incubation time was significant for 500 µg/ml of A. herba-alba Asso extract. Conclusion: Significant in vitro anti-sickling activity of A. herba-alba Asso extract was demonstrated in RBCs pretreated with 2% sodium metabisulfite. The results obtained in this study have shown significant in vitro anti-sickling activity of A. herba-alba Asso extract, and these findings may justify the use of this plant in the management of SCD. Keywords: In vitro, Anti-sickling activity, Artemisia herba-alba Asso, Emmel test, Sodium metabisulfite, Percentage, Unsickled red blood cells, Sickle cell disease. INTRODUCTION Sickle cell disease (SCD) is an inherited genetic disorder that affects the hemoglobin within the red blood cells (RBCs). The recurrent pain and complications caused by the disease can interfere with many aspects of the patient’s life including education, employment, and psychosocial development. The sickle cell trait is now known to be widespread, reaching its highest prevalence in parts of Africa as well as among people in the Mediterranean basin and Saudi Arabia [1]. Artemisia herba-alba Asso (Asteraceae family), commonly known as white wormwood or desert wormwood (Arabic name chih), is a grayish-strongly aromatic dwarf shrub native to the South Western Europe, Northern Africa, Arabian Peninsula, and Western Asia [2]. The sesquiterpene lactones compounds are the main product which can be obtained from the A. herba-alba Asso which give its medical and pharmaceutical important [3]. The pharmacological activities of A. herba-alba Asso extract are mentioned by various researchers such as antidiabetic effect [4-7], antimicrobial activity [8], antifungal activities [8,9], and antioxidant effect [9-12]. In this study, we try to find out the anti-sickling effect of methanolic extracts of A. herba-alba Asso for reducing complicated management and cost-effective treatment of sickle cell patient. METHODS Preparation of methanolic extract of plants In this study, A. herba-alba Asso (aerial parts) was collected from Al-Qassim region in the North central part of Saudi Arabia in June 2014. A voucher sample is stored at the Department of Medical Laboratories, Majmaah University . The dried plant sample was ground in a blender with a particular size to ensure the powder in identical size, and then AQ1 100 g of the powder was soaked for 5-7 days with 1000 ml of 80% methanol at 25°C. After filtration, the filtrate was evaporated with a rotary evaporator to remove the methanol under reduced pressure at 50°C. The dry crude extract of the plant samples was stored in the refrigerator in a dark glass bottle until use. A stock solution 0.1 g/ml from the crude extract was prepared by dissolving 0.1 g of dry crude extract in 1 ml (dimethylsulfoxide [DMSO]) and then diluted in 9 ml normal saline; this stock solution was stored in a refrigerator for 5 days until use. Collection of blood samples The blood samples used in the evaluation of the anti-sickling activity of the plant extract in this study were taken from patients known to have SCD, attending in the King Khaled Hospital in Majmaah. All these patients were confirmed regarding their SS status using hemoglobin electrophoresis test. The blood samples were collected in sodium ethylenediamine tetraacetic acid (EDTA) tubes and stored for maximum a few hours for the experiment. A written informed consent was read and signed by all the patients participating in the study. All research procedures have been approved by the National Ethical Committee, King Abdulaziz for Science and Technology, Kingdom of Saudi Arabia, approval number: MUREC-Jan.06/COM-2015. Anti-sickling activity Washing of RBCs About 4 ml EDTA blood samples obtained from patients were centrifuged at 3000 rpm for 10 minutes to remove the plasma. The resulting packed erythrocytes were washed 3 times with 1 ml sterile normal saline per 5 ml of blood. The samples were then centrifuged each time to remove the supernatant. Washed RBC was then re-suspended in remaining suspension and used for the analysis. Research Article AJPCR_12450_RA