5th International Seminar on Pharmaceutical Science and Technology (ISPST)-3rd International Seminar and Expo on Jamu-13th Annual ISCC 2022. | 127 NEUROPROTECTIVE AND ANTIOXIDANT ACTIVITIES OF AQUEOUS EXTRACT MORINGA OLEIFERA LEAVES Original Article EMNI PURWONINGSIH 1,2* , WAWAIMULI AROZAL 3 , HEE J. LEE 4 , ABDUL MUNIM 5 1 Doctoral Program in Biomedical Science Faculty of Medicine University of Indonesia. Jl. Salemba Raya no. 6, Kenari, Kec. Senen, Jakarta Pusat, DKI Jakarta, Indonesia 10430, 2 Faculty of Medicine, Universitas Muhammadiyah Sumatera Utara. Jl. Gedung Arca no.53, North Sumatera 20217, 3 Department of Pharmacology and Therapeutics, Faculty of Medicine, University of Indonesia, Jl. Salemba Raya no. 6, Kenari, Kec. Senen, Jakarta Pusat, DKI Jakarta, Indonesia 10430, 4 Kangwon National University, South Korea, 5 Faculty of Pharmacy, University of Indonesia, Jl. Prof. Dr. Mahar Mardjono, Pondok Cina, Beji, Depok City, West Java, Indonesia 16424 * Email: emni.purwoningsih@umsu.ac.id; heejaelee@kangwon.ac.kr Received: 29 Sep 2022, Revised and Accepted: 12 Nov 2022 ABSTRACT Objective: To investigate the neuroprotective and antioxidant effects of leaves aqueous extract Moringa oleifera (MW) in chronic stress mouse models. Methods: Water immersion and stress restraint for 16 d to obtain a chronic stress model animal. Moringa extract flour dissolved in Aquades, dose 800 mg/kg for 23 d, for chronic Stress+MOW group. Fluoxetine in aquades at a dose of 18 mg/kg BW for 23 d for chronic stress group+Fluoxetine. Aquades were given to normal mice (group N), and mice under chronic stress conditions (chronic stress control group). Furthermore, measure behavioral abnormalities by testing depressive behavior and oxidative stress parameters such as anxiety, Brain-derived neurotrophic factors (BDNF). Results: Moringa oleifera water extract administration can improve behavioral disorders caused by stress by decreasing immobility time on the Force swim test, increasing time in the middle area, and increasing the number of returns to center areas on the Open field test. When chronically stressed mice were given fluoxetine and MOW, their MDA levels (p=0.008 and 0.041, respectively) and SOD activity (p=0.001 and 0.004) decreased significantly compared to the chronic stress control group. In contrast, Catalase activity increased significantly in chronically stressed mice given fluoxetine and MOW compared to the chronic stress control group (p=0.010 and 0.013). Administration of fluoxetine and MOW may increase the expression of mRNA BDNF compared to the chronic stress control group (p=0.000 and 0.013). Conclusion: The study found that MOW can improve behavioral abnormalities, namely anxiety and depression behavior caused by chronic stress exposure, through antioxidant pathways and oxidant systems, and also BDNF Keywords: Moringa oleifera, Anxiety, Oxidative stress, Antioxidant, Chronic stress © 2022 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/) DOI: https://dx.doi.org/10.22159/ijap.2022.v14s5.26 Journal homepage: https://innovareacademics.in/journals/index.php/ijap INTRODUCTION The most widely distributed neurotrophin, brain-derived neurotrophic factor (BDNF), and oxidative stress (OS) may be important in several pathological manifestations of neurodegenerative disorders [1]. The brain-derived neurotrophic factor (BDNF) plays a significant role in various stress-related mood disorders [2]. Stress-related responses have also been altered in animal studies of BDNF deficit in vivo, and BDNF is a common downstream intermediary for various exposures that potentiate anxiety-and depressive-like behavior [3]. Chronic stress disrupts prooxidant-antioxidant stability, favoring oxidative reactions. In response to cellular metabolites, ROS causes the creation of peculiar free radical oxidation products [4]. The brain is thought to be particularly vulnerable to oxidative stress or redox imbalances due to its high oxygen consumption and an environment rich in lipids. Thus, it should be no surprise that oxidative stress is linked to a number of brain disorders, including neurodegenerative diseases, psychiatric conditions, and anxiety [5]. Previous literature has demonstrated M. oleifera positive benefits, including neuroprotective and antioxidant properties [6]. Moringa oleifera leaves are enriched in phytochemicals such as tannins, sterols, saponins, terpenoids, phenolics, alkaloids, and flavonoids [7]. Previous rodent studies found that extracts of Moringa oleifera leaves have anxiolytic and antiepileptic properties [8, 9], antidepressant [10], and improves memory [11]. MATERIALS AND METHODS Materials The Moringa leaves powder was obtained from PT Javaplant (Solo, Indonesia); it was made by an aqueous extraction method and filled in with maltodextrin. Superoxide Dismutase (SOD) Colorimetric Activity Kit (ab65354, colorimetry, Abcam, Cambridge, UK), catalase activity assay kit (colorimetric, ab83464, Abcam, Cambridge, UK), and Malondialdehyde Lipid Peroxidation (MDA) Assay Kit (MyBiosource, Inc. San Diego, USA, 822354). Animal preparation We purchased 24 male BALB/c mice (25-30 g) from Biopharma Laboratory and Animal Breeding, Bandung-Indonesia. The mice were kept in cages in a closed system at 24±2 °C and a 12 h light/dark cycle and allowed access to food and drink ad libitum. The rats were randomly divided into four groups of five. Group Normal: normal mice received aquadest. Group stress chronic control: mice with induction of water immersion and restraint stress. Group stress chronic+fluoxetine: mice models chronic stress+administration of fluoxetine (18 mg/kg BW/d/oral) Group stress chronic+MOW: stress chronic mice models+administration of Moringa water extract 800 mg/kg BW/d/oral (MOW). Administration of fluoxetine and Moringa for 23 d of the experiment. Stress protocol The combination of water immersion and restraint stress for 16 d was used in this study to create a stress animal model. The combination of restraint stress and water immersion stress, combining psychological and physical stressors, can further cause depressive symptoms [12]. Chronic stress uses the method described by Yasugaki et al., with some modifications. Mice are inserted into a 50 ml conical polypropylene centrifuge tube with several holes for air circulation [10]. Moreover, the tube is immersed in water (the temperature at 22 °C) in a vertical position up to the xiphoid limit. International Journal of Applied Pharmaceutics ISSN- 0975-7058 Vol 14, Special Issue 5, 2022