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Evaluation of Nootropic Activity of Limonia Acidissima Against Turk J Pharm Sci 2021;18(1):3-9 DOI: 10.4274/tjps.galenos.2019.30316 ORIGINAL ARTICLE Evaluation of Nootropic Activity of Limonia acidissima Against Scopolamine-induced Amnesia in Rats Limonia acidissima’nın Sıçanlarda Skopolamin ile İndüklenen Amneziye Karşı Nootropik Aktivitesinin Değerlendirilmesi Kailas K MALI1*, Guruprasad V SUTAR2, Remeth J DIAS3, Omkar A DEVADE1 1Adarsh College of Pharmacy, Department of Pharmacology, Vita, Maharashtra, India 2Annasaheb Dange College of B-Pharmacy, Department of Pharmacology, Astha, Maharashtra, India 3Government College of Pharmacy, Karad, Maharashtra, India ABSTRACT Objectives: The present study aimed to evaluate the nootropic activity of Limonia acidissima in rats. Materials and Methods: Methanolic extract of Limonia acidissima was used to evaluate nootropic activity, piracetam (200 mg/kg, i.p.) was used as a standard, and scopolamine (1 mg/kg, i.p.) was used to induce amnesia. The effect of drugs on learning and memory in rats was evaluated by using the Y-maze task and elevated plus maze on scopolamine-induced amnesia models. Locomotor activity was performed using an actophotometer. Also, levels of acetylcholinestrease, including histopathological examination of rat brains, were assessed. Results: Methanolic extract of Limonia acidissima showed increased alteration of the behavior response and percentage spontaneous alteration with the Y-maze task. In the elevated plus maze scopolamine-induced amnesia model, methanolic extract of Limonia acidissima showed a decrease in transfer latency, which is indicative of cognition improvement. Methanolic extract increased locomotor activity in rats and decreased the levels of acetylcholinestrease enzyme significantly. A histopathological study with both low and high doses of extract showed effective regenerative scores as compared to normal control, negative control and standard treatment. Conclusion: The results suggested that the administration of methanolic extract of Limonia acidissima enhances learning and memory in different experimental models. The histopathological study revealed the neuroprotective property of the extract. The study indicates that the extract may be used in the treatment of Alzheimer’s disease. Key words: Nootropic activity, Limonia acidissima, Alzheimer’s disease, piracetam, scopolamine ÖZ Amaç: Bu çalışmada, sıçanlarda Limonia acidissima’nın nootropik aktivitesinin değerlendirilmesi amaçlanmıştır. Gereç ve Yöntemler: Nootropik aktiviteyi değerlendirmek için Limonia acidissima’nın metanol ekstresi, standart olarak pirasetam (200 mg/kg, i.p.) ve amneziyi indüklemek için skopolamin (1 mg/kg, i.p.) kullanıldı. İlaçların sıçanlarda öğrenme ve hafıza üzerindeki etkisi, skopolamin ile indüklenen amnezi modelinde Y-labirent testi ve yükseltilmiş artı labirent testi kullanılarak değerlendirildi. Lokomotor aktivite, bir aktofotometre kullanılarak gerçekleştirildi. Ayrıca, sıçan beyinlerinin asetilkolinestreaz aktivitesinin değerlendirilmesi de dahil olmak üzere histopatolojik incelemesi yapıldı. Bulgular: Y-labirent testi ile Limonia acidissima’nın metanol ekstresinin davranış tepkisinde ve yüzde spontan değişikliklerde artışa neden olduğu gösterildi. Skopolamin ile indüklenen amnezi modelinin kullanıldığı yükseltilmiş artı labirent testinde, Limonia acidissima’nın metanol ekstresinin, biliş gelişiminin göstergesi olan transfer gecikmesinde bir azalmaya yol açtığı gösterildi. Metanol ekstresi, sıçanlarda lokomotor aktiviteyi artırdı ve asetilkolinestreaz enzim aktivitesini önemli ölçüde düşürdü. Hem düşük hem de yüksek dozda ekstreyle yapılan histopatolojik çalışmada, normal kontrol, negatif kontrol ve standart tedaviye kıyasla efektif rejeneratif skorlar elde edildi. Sonuç: Sonuçlar, Limonia acidissima’nın metanol ekstresinin uygulanmasının farklı deneysel modellerde öğrenmeyi ve hafızayı geliştirdiğini göstermiştir. Histopatolojik çalışma, ekstrenin nöroprotektif özellikte olduğunu ve ekstrenin Alzheimer hastalığının tedavisinde kullanılabileceğini göstermiştir. Anahtar kelimeler: Nootropik aktivite, Limonia acidissima, Alzheimer hastalığı, pirasetam, skopolamin *Correspondence: [email protected], Phone: +09552527353 ORCID-ID: orcid.org/0000-0002-1789-3592 Received: 13.12.2018, Accepted: 21.03.2019 ©Turk J Pharm Sci, Published by Galenos Publishing House. 3 4 MALI et al. Nootropic Activity of Limonia acidissima INTRODUCTION Drugs and chemicals According to the World Health Organization, approximately 450 Piracetam (Dr. Reddy’s, India), scopolamine hydrobromide (APP million people suffer from a mental or behavioral disorder.1 Pharmaceuticals, India), and normal saline were used for the Dementia (age-related mental disorder) is a characteristic study. All other reagents and chemicals were of analytical symptom of Alzheimer’s disease (AD).2-4 AD is a progressive, grade and procured from Loba Chemie, Mumbai, India. neurodegenerative, and cerebrovascular disease.2,5 It destroys Plant material and preparation of extracts cells in the brain, causing problems with memory, unusual Plant material was collected from Phaltan, identified, and behavior6, difficulty thinking, personality changes,7 and authenticated by the Department of Botany, Yashwantrao ultimately death.8-10 AD is characterized by the loss of neuronal Chavan Institute of Science, Satara, Maharashtra, India. A cells and is primarily linked to neurofibrillary tangles and voucher specimen (no: 57) of plant material was kept in the neuritic plaques.7,11 The cholinergic system in the brain plays Department of Pharmacognosy, Yashoda Technical Campus, an important role in learning and memory,12 which involves Satara. The fully ripe fruits and leaves of the plant were dried acetylcholine (Ach).13 Dementia is produced due to reduction in the shade for 1 week. Then, fruits were cracked open, and of Ach in the brains of patients with AD.7 In rodents and human the rind, seed, and pulp were separated and shade dried with beings, drugs like scopolamine impair learning and memory.4,13 the leaves at room temperature for more than 2 weeks. After Memory loss, amnesia, dementia, anxiety, schizophrenia, and drying, the rind, pulp, and seeds were ground separately into AD may be produced due to certain conditions like age, stress, a coarse powder, and the leaves was also separately ground and emotion.14,15 into a coarse powder for further extraction. The coarsely powdered material was extracted with methanol by using the There are a few nootropic medicines used in the treatment of AD, Soxhlet extraction method for 4 days at 45°C, at a ratio of 1:1:1 called nootropic drugs, belonging to the class of psychotropic (seeds, pulp, and leaves). The obtained extract was filtered and agents.16 The term nootropic was coined by Giurgea in 1972, from concentrated (yield 8.8%). the Greek noon (mind) and tropos (turn).17 Nootropics are also referred to as smart drugs, as they improve mental functions Preliminary phytochemical investigation such as memory, increase blood circulation to the brain, and The methanolic extract was subjected to phytochemical tests improve the oxygen supply to the brain.18 Synthetic medicines for alkaloids, flavonoids, glycosides, saponins, carbohydrates, like tacrine, dopnezil,19 aniracetam, piracetam, and rivastigmine and tannins.26 are used for the treatment of cognitive dysfunction and memory Acute toxicity study loss associated with AD.20 However, these drugs pose some An acute toxicity study was performed according to OECD adverse effects and bioavailability issues.21 To overcome these guideline 423. Six swiss albino female mice, weighing in the problems, researchers are seeking herbal formulations that can range of 20-25 g, were administered the test solution at a dose overcome the adverse effects of synthetic drugs. of 2.000 mg/kg. After administration of the test formulation at Many herbs have been studied extensively and reported to have intervals of 30 min, 1 h, 2 h, 4 h, 24 h, 48 h, and 72 h up to a memory-enhancing properties.22 The plant Limonia acidissima period of 14 days, mice were observed for clinical signs, gross is a herbal drug used in Ayurvedic systems of medicine.23 It behavioral changes, and mortality.27 is also used in a variety of conditions for its antimicrobial, Behavioral study hepatoprotective, antidiarrhoeal, anticancer, diuretic, hepatoprotective, antispermatogenic, antioxidant, antidiabetic, Rats were trained for the behavioral study by conducting 1 week of training during which they did not receive any plant extract or and wound-healing activities.24,25 drug. The completely trained rats were selected for the study. The present study seeks to determine whether the methanolic An experiment was carried out in the light period between 8.00 extract of leaves and fruit pulp of Limonia acidissima shows am and 03:00 pm in a sound- proof room. nootropic activity in an animal model. Y-maze task MATERIALS AND METHODS Five groups of animals were formed, each group comprising six animals as follows. Group 1: Control group (normal saline), Experimental animals group 2: Negative group (scopolamine 1 mg/kg i.p), group 3: Wistar rats of either sex weighing between 150 and 200 g were Standard treatment (piracetam 200 mg/kg i.p. + scopolamine used for the present study. They were housed under standard 1 mg/kg i.p.), group 4: Low dose of extract + scopolamine laboratory conditions (temperature 25°C±1°C), relative humidity (200 mg/kg p.o + 1 mg/kg i.p.), group 5: High dose of extract + 55%±5% and 12.00:12.00 h dark: light cycle) with a standard scopolamine
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