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Evaluation of the Erythrocyte Fragility, Haematological Parameters And Alinteri J. of Agr. Sci. (2020) 35(1): 22-28 http://dergipark.gov.tr/alinterizbd e-ISSN: 2587-2249 http://www.alinteridergisi.com/ [email protected] DOI: 10.28955/alinterizbd.740369 RESEARCH ARTICLE Evaluation of the Erythrocyte Fragility, Haematological Parameters and Antioxidant Properties of Platanus orientalis Leaf Infusion Against Ethanol Toxicity in Rats Abdulahad Doğan1* • Fatih Dönmez1 • Özgür Ozan Anuk2 1Van Yüzüncü Yıl University, Faculty of Pharmacy, Department of Biochemistry, Van/Turkey 2Van Yüzüncü Yıl University, Institute of Health Sciences, Department of Biochemistry, Van/Turkey ARTICLE INFO ABSTRACT Article History: The aim of this study was to evaluate the protective effects of the leaf infusion of chinar (Platanus Received: 13.11.2019 orientalis L.) on erythrocyte fragility, haematological parameters and antioxidant status against Accepted: 01.03.2020 ethanol-induced oxidative stress in rats. Thirty male rats were divided into five groups: Control, Available Online: 20.05.2020 Ethanol, Ethanol+Silymarin (10 mg kg-1), Ethanol+PO-20 mg mL-1 infusion, and Ethanol+PO-60 mg mL- 1 Keywords: infusion. According to the results, in the Ethanol group, erythrocyte counts, red cells distribution, plateletcrit, platelet and lymphocyte levels significantly decreased compared to the Control group, Platanus orientalis while PO-60 dose-fed group showed a significant increase in haematocrit and haemoglobin values Haematological Parameters compared to the Ethanol group. There were significant changes in erythrocyte fragility of Ethanol Erythrocyte Fragility and Ethanol-treatment groups at different NaCl concentrations of 0.3 and 0.6 according to Control Antioxidant group. It was observed that PO Leaf infusion reduced the hemolysis caused by ethanol at a Ethanol concentration of 0.3% NaCl, thus reducing the values to the control values. In addition, PO leaf Rat infusion caused a significant increase in total antioxidant status against ethanol toxicity and a significant decrease in total oxidative status and oxidative stress index. It was concluded that PO leaf infusion may have antihematotoxic effect, reducing erythrocyte fragility and increase antioxidant capacity against ethanol toxicity. Please cite this paper as follows: Doğan, A., Dönmez, F. and Anuk, Ö. O. (2020). Evaluation of the Erythrocyte Fragility, Haematological Parameters and Antioxidant Properties of Platanus orientalis Leaf Infusion Against Ethanol Toxicity in Rats. Alinteri Journal of Agriculture Sciences, 35(1): 22- 28. doi: 10.28955/alinterizbd.740369 Introduction dehydrogenase, aldehyde dehydrogenase and catalase (Zakhari, 2006; Pari and Suresh, 2008). The activation of O2 by Reactive oxygen species (ROS) or free radicals are .- cytochromes P450 generates superoxide (O2 ) and hydrogen substances generated by oxygen (O2) metabolism which is peroxide (H2O2), which contributes to oxidative tissue damages balanced by the proportion of oxidant formation and the (Lu and Cederbaum, 2008). Free radicals are among the by- proportion of oxidant elimination (Sinha and Dabla, 2015). products of EtOH metabolism and are known to cause cellular Oxidative stress (OS) results in the formation of free radicals and tissue damage, unless the body can use antioxidants to or ROS due to insufficient antioxidant defence systems. There clean them up (Varga et al., 2017). Ethanol-induced oxidative are many endogenous and exogenous factors causing OS. stress increases the production of ROS which cause damage to Ethanol (EtOH) is one of the exogenous factors causing OS. The the RBC membrane via lipid peroxidation (Wrońska‐Noferδ et majority of EtOH is metabolised in the liver by alcohol al., 1991). Additionally, it causes a deteriorating effect on the * Corresponding author E-mail address: [email protected] 22 Doğan, Dönmez and Anuk (2020). Alınteri Journal of Agriculture Sciences 35(1): 22-28 membrame integrity by increasing fluidity as well as the in the dark until dryness. The powdered samples were kept in deactivation of membrane-bound receptors and enzymes boiling water (100 °C) for about 2 min. Then, the heating was (Arihan et al., 2016). stopped and the ground leaves were allowed to remain in the water for about 15 min. Subsequently, the liquid in the To the best of our knowledge, the effects of PO leaf container was first filtered through a gauze cloth (rough-hew) infusion on hematological parameters have not been and then through a 0.45-µm hydrophilic filter (Millipore) using invesitgated so far. It was one of our main goals to uncover the a disposable injector. possible effects of PO leaf infusion consumption in eliminating ethanol-induced hematological problems. Since, Chemicals haematological parameters, erythrocytes, leukocytes and Ethanol, silymarin, adenosine, hydrogen peroxide solution platelets, are commonly used to diagnose various diseases such (H O , %30), ammonium sulphate, phenol, o-dianisidine, as anemia, inflammatory, autoimmune, allergic, infectious, 2 2 ethylenediaminetetraacetic acid (EDTA), sodium hypochlorite cancer, neutrophil and lymphocyte-related parameters as well solution, potassium dihydrogen phosphate (KH PO ), sodium as some further common disorders (Gao et al., 2019). The 2 4 chloride (NaCl), and sodium dihydrogen citrate anhydrous osmotic fragility test is one of the additional screening tests (C H NaO ) were supplied by Sigma Chemical Co. (St. Louis, for evaluates the red blood cells ability to associate hypotonic 6 7 7 MO, USA). TAS and TOS kits were supplied by Rel Assay solutions (King et al., 2015). The excess of hemolysis means Diagnostics Laboratories Ltd. that the structure of the cell membrane is destroyed. Antioxidant mechanisms protect cells and tissues against Animals free radicals. The total oxidative status (TOS), total anti- The male rats (Wistar albino) of ~ 2 months of age and an oxidative status (TAS) ratio and oxidative stress index (OSI) are average weight of 200 g were provided by the Experimental indicative parameters of the degree of OS (Tamura et al., Animal Research Centre, Van Yuzuncu Yil University (Van, 2016). TAS and TOS are well known indicator parameters for Turkey). The animals were housed at 25 ± 2 °C at a daily the analysis of the oxdative stress in many diseases such as light/dark photoperiod of 10:14. All of the animals received cancer, diabetes, cardiovascular and organ damage. water and a wheat-soybean-based diet ad libitum in stainless Platanus orientalis (Plantanaceae), also known as Chinar or steel cages, and received humane care according to the Oriental plane, is commonly used in folk medicine against criteria outlined in the ‘Guide for the Care and Use of tooth and knee pain, wounds, inflammation, liver, kidney and Laboratory Animals’ prepared by the National Academy of stomach diseases (Dogan and Anuk, 2019). In previous studies Science and published by the National Institute of Health. The it has been reported that P. orientalis contains important ethic regulations were followed in accordance with national compounds such as kaempferol and kaempferol derivatives, and institutional guidelines for the protection of animal caffeic acid, platanoside, tiliroside, flavonol and welfare during experiments. This study was approved by the proanthocyanidin glycosides, phytol derivatives, Ethics Committee of Van Yuzuncu Yil University (Protocol benzaldehyde, palmitic acid, 2,4-ditert-butylphenol, stearic number: 27552122-604.01.02-E.70881). acid, octadecanoic acid, linoleic acid, and linolenic acid Experimental Design (Dogan and Anuk, 2019; Khan, 2017). After toxicity test, the rats were randomly divided into 5 The purpose of this study was to determine the possible groups, with each containing 6 rats and the study was effects of PO-leaf infusion on the erythrocyte fragility, continued for 28 days. haematological parameters and antioxidant/oxidant capacity in erythrocyte of rats with ethanol-induced oxidative stress. Control group: The rats were allowed to receive tap water and a standard pellet diet ad libitum. Materials and Methods Ethanol group: The rats were allowed to receive 20% Plant Material and Preparation of the Infusion ethanol and a standard pellet diet ad libitum. The dose of ethanol was selected on the basis of a 20% concentration that The Platanus orientalis leaves were collected by Abdulahad was administered orally, which caused OS (Dogan and Anuk, Dogan in Haci Hamza hamlet, district of Dargeçit, city of 2019). Mardin, in the south-eastern Anatolian region of Turkey (GPS Ethanol + Silymarin: The rats were allowed to receive 20% coordinates: 37°33′19.7″N; 41°47′43.3″E) in August, 2017. The ethanol and silymarin (10 mg kg-1, single dose per day) and identification of the samples was confirmed by Dr. Abdullah were treated orally during the experimental period. Dalar at the Department of Pharmaceutical Botany, Faculty of Pharmacy, Van Yuzuncu Yil University, Turkey, and a voucher Ethanol + PO-20 group: The rats were allowed to receive specimen was deposited in the university’s herbarium 20% ethanol and P. orientalis (20 mg mL-1) leaf infusion during (Herbarium code: 340 and Collector No: A.D-761, Van Yuzuncu the experimental period. Yil University Faculty of Pharmacy Herbarium). Ethanol + PO-60 group: The rats were allowed to receive The infusion of PO-leaf was prepared according to the study 20% ethanol and P. orientalis (60 mg mL-1) leaf infusion during by Dogan and Anuk (2019). Briefly, the fresh PO- leaf samples the experimental period. were washed under tap water and dried
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