AAD  Vol.5 No.2 , June 2016
Neuroprotective Effect of Phyllanthus acidus L. on Learning and Memory Impairment in Scopolamine-Induced Animal Model of Dementia and Oxidative Stress: Natural Wonder for Regulating the Development and Progression of Alzheimer’s Disease
Abstract: Nature is the best source of complementary and alternative medicine. The plant Phyllanthus acidus (PA) L. has been used traditionally in pain, inflammatory and oxidative stress related disorders. In this consequence, methanolic extract of PA (MEPA) was selected to explore the ability of this plant to enhance cognitive function, brain antioxidant enzymes and anti-acetylcholinesterase activity which can be used for the treatment of oxidative stress related disorders like Alzheimer’s disease (AD). The purpose of this study was to investigate the neuroprotective effect of MEPA on learning and memory impairment in scopolamine-induced rats of dementia and oxidative stress. Treatment with MEPA (i.e., 100 and 200 mg/kg b.w.) was investigated in scopolamine-treated Swiss albino male rats for 14 days and its neuroprotective effects were examined using Elevated Plus Maze (EPM) test, Passive Avoidance (PA) test, Novel Object Recognition (NOR) test, Morris Water Maze (MWM) test as well as level of antioxidant enzymes such as catalase (CAT), super oxide dismutase (SOD), glutathione reductase (GSR), glutathione-S-transferase (GST), reduced glutathione (GSH), glutathione peroxidase (GSH-Px), lipid peroxidation (TBARS) contents and acetylcholinesterase (AChE) activity in rat brain tissue homogenates. Administration of MEPA significantly (P < 0.05, P < 0.01; P < 0.01) decreased RTL (retention transfer latency) in rats on 7th and 14th day compared to the disease control and control group in the EPM test. In PA test the doses of MEPA suggestively (P < 0.05, P < 0.001; P < 0.05, P < 0.01) increased STL (step-through latency) in rats on 7th and 14th day with respect to disease control and control group. For NOR test administration of MEPA considerably (P < 0.01, P < 0.001; P < 0.01) increased the DI (discrimination index) in rats with respect to that of disease control and control group. The doses of MEPA markedly (P < 0.05, P < 0.01; P < 0.01) decreased EL (escape latency) and significantly (P < 0.01, P < 0.001; P < 0.05, P < 0.01) increased TSTQ (time spent in the target quadrant) on successive days as compared to that of disease control and control group in the acquisition trial of MWM test. In case of probe trial of MWM test MEPA administration considerably (P < 0.01; P < 0.05, P < 0.01) increased TSTQ and significantly (P < 0.05, P < 0.01; P < 0.05, P < 0.01) increased TSA (time spent in the annuli) in rats on successive days as compared to that of disease control and control group. MEPA administration significantly (P < 0.05, P < 0.01, P < 0.001; P < 0.05, P < 0.01) increased the level of CAT, SOD, GSR, GST GSH, GSH-Px and markedly (P < 0.01; P < 0.01, P < 0.001) decreased TBARS level through inhibiting lipid peroxidation as well as significantly (P < 0.01, P < 0.001; P < 0.05, P < 0.01, P < 0.001) decreasing AChE activity in rats brain compared to the disease control and control group. The present study demonstrates that MEPA showed the neuroprotective effect by improving cognitive functions and reduces oxidative stress by increasing the level of brain antioxidant enzymes as well as decreasing lipid peroxidation and acetylcholinesterase activity. Therefore, this plant extract can be used for enhancing learning, memory, antioxidant potentiality and anti-acetylcholinesterase activity in neurodegenerative disorders like AD.
Cite this paper: Uddin, M. , Mamun, A. , Hossain, M. , Ashaduzzaman, M. , Noor, M. , Hossain, M. , Uddin, M. , Sarker, J. and Asaduzzaman, M. (2016) Neuroprotective Effect of Phyllanthus acidus L. on Learning and Memory Impairment in Scopolamine-Induced Animal Model of Dementia and Oxidative Stress: Natural Wonder for Regulating the Development and Progression of Alzheimer’s Disease. Advances in Alzheimer's Disease, 5, 53-72. doi: 10.4236/aad.2016.52005.

[1]   Sternberg, R.J. and Pretz, J.E. (2005) Cognition and Intelligence: Identifying the Mechanisms of the Mind. Cambridge University Press, New York.

[2]   Selnes, O.A. and Vinters, H.V. (2006) Vascular Cognitive Impairment. Nature Clinical Practice Neurology, 2, 538- 547.

[3]   Sosa, A., Albanese, E., Stephan, B.C.M., Dewey, M., Acosta, D., Ferri, C.P., et al. (2012) Prevalence, Distribution, and Impact of Mild Cognitive Impairment in Latin America, China, and India: A 10/66 Population-Based Study. PLoS Medicine, 9, 1-11.

[4]   Querfurth, H.W. and LaFerla, F.M. (2010) Alzheimer’s Disease. New England Journal of Medicine, 362, 329-344.

[5]   Wimo, A., Winblad, B., Aguero-Torres, H. and von Strauss, E. (2003) The Magnitude of Dementia Occurrence in the World. Alzheimer Disease & Associated Disorders, 17, 63-67.

[6]   Asaduzzaman, M., Uddin, M.J., Kader, M.A., Alam, A.H.M.K., Rahman, A.A. and Rashid, M. (2014) In Vitro Acetylcholinesterase Inhibitory Activity and the Antioxidant Properties of Aegle marmelos Leaf Extract: Implications for the Treatment of Alzheimer’s Disease. Psychogeriatrics, 14, 1-10.

[7]   Chang, Y.T., Chang, W.N., Tsai, N.W., Huang, C.C., Kung, C.T. and Su, Y.J. (2014) The Roles of Biomarkers of Oxidative Stress and Antioxidant in Alzheimer’s Disease: A Systematic Review. BioMed Research International, 2014, 1-11.

[8]   Oh, J.H., Choi, B.J., Chang, M.S. and Park, S.K. (2009) Nelumbo nucifera Semen Extract Improves Memory in Rats with Scopolamine-Induced Amnesia through the Induction of Choline Acetyltransferase Expression. Neuroscience Letters, 461, 41-44.

[9]   Fan, Y., Hu, J., Li, J., Yang, Z., Xin, X., Wang, J., et al. (2005) Effect of Acidic Oligosaccharide Sugar Chain on Scopolamine-Induced Memory Impairment in Rats and Its Related Mechanisms. Neuroscience Letters, 374, 222-226.

[10]   El-Sherbiny, D.A., Khalifa, A.E., Attia, A.S. and Eldenshary, E.S. (2003) Hypericum perforatum Extract Demonstrates Antioxidant Properties against Elevated Rat Brain Oxidative Status Induced by Amnestic Dose of Scopolamine. Pharmacology Biochemistry and Behavior, 76, 525-533.

[11]   Jeong, E.J., Lee, K.Y., Kim, S.H., Sung, S.H. and Kim, Y.C. (2008) Cognitive-Enhancing and Antioxidant Activities of Iridoid Glycosides from Scrophularia buergeriana in Scopolamine-Treated Mice. European Journal of Pharmacology, 588, 78-84.

[12]   Sultana, R. and Butterfield, D.A. (2010) Role of Oxidative Stress in the Progression of Alzheimer’s Disease. Journal of Alzheimer’s Disease, 19, 341-353.

[13]   Hossain, M.S., Asaduzzaman, M., Uddin, M.S., Noor, M.A.A., Rahman, M.A. and Munira, M.S. (2015) Investigation of the in Vitro Antioxidant and Cytotoxic Activities of Xanthosoma sagittifolium Leaf. Indo American Journal of Pharmaceutical Research, 5, 3300.

[14]   Kuhla, B., Haase, C., Flach, K., Luth, H.J., Arendt, T. and Munch, G. (2007) Effect of Pseudophosphorylation and Cross-Linking by Lipid Peroxidation and Advanced Glycation End Product Precursors on Tau Aggregation and Filament Formation. Journal of Biological Chemistry, 282, 6984-6991.

[15]   Zimmerman, G. and Soreq, H. (2006) Termination and Beyond: Acetylcholinesterase as a Modulator of Synaptic Transmission. Cell and Tissue Research, 326, 655-669.

[16]   Lane, R.M., Potkin, S.G. and Enz, A. (2006) Targeting Acetylcholinesterase and Butyrylcholinesterase in Dementia. International Journal of Neuropsychopharmacology, 9, 101-124.

[17]   Zawia, N.H., Lahiri, D.K. and Cardozo-Pelaez, F. (2009) Epigenetics, Oxidative Stress, and Alzheimer Disease. Free Radical Biology & Medicine, 46, 1241-1249.

[18]   Jayakumar, T., Thomas, P.A. and Geraldine, P. (2007) Protective Effect of an Extract of the Oyster Mushroom, Pleurotus ostreatus, on Antioxidants of Major Organs of Aged Rats. Experimental Gerontology, 42, 183-191.

[19]   Gilgun-Sherki, V., Melamed, E. and Offen, D. (2001) Oxidative Stress Induced-Neurodegenerative Diseases: The Need for Antioxidants That Penetrate the Blood Brain Barrier. Neuropharmacology, 40, 959-975.

[20]   Uttara, B., Singh, A.V., Zamboni, P. and Mahajan, R.T. (2009) Oxidative Stress and Neurodegenerative Diseases: A Review of Upstream and Downstream Antioxidant Therapeutic Options. Current Neuropharmacology, 7, 65-74.

[21]   Sharma, J., Chawla, R., Kumar, R., Sharma, A., Sharma, R.K. and Arora, R. (2013) Camellia sinensis as a Safe Neuroprotective Radiation Counter Measure Agent. International Journal of Pharmaceutical Science Invention, 2, 26-33.

[22]   Duraipandiyan, V., Ayyanar, M. and Ignacimuthu, S. (2006) Antimicrobial Activity of Some Ethnomedicinal Plants Used by Paliyar Tribe from Tamil Nadu, India. BMC Complementary and Alternative Medicine, 6, 35.

[23]   Jamison, D.T., Breman, J.G. and Meashametal, A.R. (2006) Complementary and Alternative Medicine, in Disease Control Priorities in Developing Countries. World Bank, Washington DC.

[24]   Annonymus (2014) Traditional Medicine.

[25]   Oken, B.S., Storzbach, D.M. and Kaye, J.A. (1995) The Efficacy of Ginkgo biloba on Cognitive Function in Alzheimer Disease. Archives of Neurology, 55, 1409-14015.

[26]   Goswami, S., Saoji, A., Kumar, N., Thawani, V., Tiwari, M. and Thawani, M. (2011) Effect of Bacopa monnierion Cognitive Functions in Alzheimer’s Disease Patients. International Journal of Collaborative Research on Internal Medicine & Public Health, 3, 285-293.

[27]   Skolnick, A.A. (1997) Old Chinese Herbal Medicine Used for Fever Yields Possible New Alzheimer Disease Therapy. Journal of the American Medical Association, 277, 776.

[28]   Habib, M.R., Rahman, M.M., Mannan, A., Zulfiker, A.H.M., Uddin, M.E. and Sayeed, M.A. (2011) Evaluation of Antioxidant, Cytotoxic, Antibacterial Potential and Phytochemical Screening of Chloroform Extract of Phyllanthus acidus. International Journal of Applied Biology and Pharmaceutical Technology, 2, 420-427.

[29]   Padmapriya, N. and Poonguzhali, T.V. (2015) Antibacterial and Antioxidant Potential of the Acetone Extract of the Fruit of Phyllanthus acidus L. International Journal of Current Research, 17, E64-E72.

[30]   Annonymus. Phyllanthus acidus.

[31]   Devi, S.S. and Paul, S.B. (2011) An Overview on Cicca acida (Phyllanthus acidus). Assam University Journal of Science & Technology: Biological and Environmental Sciences, 7, 156-160.

[32]   Catapan, E., Otuki, M.F., Viana, A.M., Yunes, R.A., Bresciani, L.F., Ferreira, J., et al. (2000) Pharmacological Activity and Chemical Composition of Callus Cultures Extracts from Selected Species of Phyllanthus. Pharmazie, 55, 945- 946.

[33]   Shilali, K., Ramachandra, Y.L., Rajesh, K.P. and Swamy, B.E.K. (2014) Assessing the Antioxidant Potential of Phyllanthus acidus Bark Extracts. International Journal of Pharmacy and Pharmaceutical Sciences, 6, 522-531.

[34]   Anjaria, J., Parabia, M., Bhatt, G. and Heals, K.R.N.A. (2002) Glossary of Selected Indigenous Medicinal Plants of India. Sristi Innovations, Ahmedabad.

[35]   Lemmens, R.H., Bunyapraphatsara, M.J. and Padua de, L.S.N. (1999) Plant Resources of South-East Asia, Medicinal and Poisonous Plants. Prosea Foundation, Bogor.

[36]   Unander, D.W., Webster, D.W. and Blumberg, B.S. (1990) Record of Usage or Assays in Phyllanthus (Euphorbiaceae) I. Subgenera Isocladus, Kirganelia, Cicca and Emblica. Journal of Ethnopharmacology, 30, 233-264.

[37]   Moniruzzaman, M., Asaduzzaman, M., Hossain, M.S., Sarker, J., Rahman S.M.A. and Rashid, M. (2015) In Vitro Antioxidant and Cholinesterase Inhibitory Activities of Methanolic Fruit Extract of Phyllanthus acidus. BMC Complementary and Alternative Medicine, 15, 403.

[38]   National Research Council (2011) Guide for the Care and Use of Laboratory Animals. National Academies Press, Washington DC.

[39]   Weon, J.B., Lee, J., Eom, M.R., Jung, Y.S. and Ma, C.J. (2014) The Effects of Loranthus parasiticus on Scopolamine-Induced Memory Impairment in Mice. Evidence-Based Complementary and Alternative Medicine, 2014, Article ID: 860180.

[40]   Jissa, G., Sai-Sailesh, K. and Mukkadan, J.K. (2014) Oral Administration of Nutmeg on Memory Boosting and Regaining in Wistar Albino Rats. Bali Medical Journal, 3, 3-10.

[41]   Saha, S. and Verma, R.J. (2015) Antioxidant Activity of Polyphenolic Extract of Phyllanthus emblica against Lead Acetate Induced Oxidative Stress. Toxicology and Environmental Health Sciences, 7, 82-90.

[42]   Organisation for Economic cooperation and Development (OECD) (2002) OECD Guidelines for the Testing of Chemicals: Acute Oral Toxicity—Acute Toxic Class Method. Paris, OECD.

[43]   Reddy, D.S. and Kulkarni, S.K. (1998) Possible Role of Nitric Oxide in the Nootropic and Antiamnesic Effects of Neurosteroids on Aging- and Dizocilpine-Induced Learning Impairment. Brain Research, 799, 215-229.

[44]   Hlinak, Z. and Krejci, I. (2002) MK-801 Induced Amnesia for the Elevated Plus-Maze in Mice. Behavioural Brain Research, 131, 221-225.

[45]   Kumar, S., Maheshwari, K.K. and Singh, V. (2009) Protective Effects of Punica granatum Seeds Extract against Aging and Scopolamine Induced Cognitive Impairments in Mice. African Journal of Traditional, Complementary and Alternative Medicines, 6, 1.

[46]   Ozkay U.D., Can, O.D., Ozkay, Y. and Ozturk, Y. (2012) Effect of Benzothiazole/Piperazine Derivatives on Intracerebroventricular Streptozotocin-Induced Cognitive Deficits. Pharmacological Reports, 64, 834-847.

[47]   Ogren, S.O., Stone, W.S. and Altman, H.J. (1987) Evidence for a Functional Interaction between Serotonergic and Cholinergic Mechanisms in Memory Retrieval. Behavioral and Neural Biology, 48, 49-62.

[48]   Van der Staay, F.J., Schuurman, T., van Reenen, C.G. and Korte, S.M. (2009) Emotional Reactivity and Cognitive Performance in Aversively Motivated Tasks: A Comparison between Four Rat Strains. Behavioral and Brain Functions, 5, 50.

[49]   Weon, J.B., Yun, B.R., Lee, J., Eom, M.R., Kim, J.S., Lee, H.E., et al. (2013) The Ameliorating Effect of Steamed and Fermented Codonopsis lanceolata on Scopolamine Induced Memory Impairment in Mice. Evidence-Based Comple- mentary and Alternative Medicine, 2013, 2-3.

[50]   Morris, R. (1984) Developments of a Water-Maze Procedure for Studying Spatial Learning in the Rat. Journal of Neuroscience Methods, 11, 47-60.

[51]   Antunes, M. and Biala, G. (2012) The Novel Objects Recognition Memory: Neurobiology, Test Procedure, and Its Modifications. Cognitive Processing, 13, 93-110.

[52]   Ennaceur, A., Neave, N. and Aggleton, J.P. (1997) Spontaneous Object Recognition and Object Location Memory in Rats: The Effects of Lesions in the Cingulate Cortices, the Medial Prefrontal Cortex, the Cingulum Bundle and the Fornix. Experimental Brain Research, 113, 509-519.

[53]   Dhingra, D. and Kumar, V. (2012) Memory-Enhancing Activity of Palmatine in Mice Using Elevated Plus Maze and Morris Water Maze. Advances in Pharmacological Sciences, 2012, Article ID: 357368.

[54]   Taati, M., Alirezaei, M., Moshkatalsadat, M.H., Rasoulian, B., Moghadasi, M. and Sheikhzadeh, F. (2011) Protective Effects of Ziziphus jujube Fruit Extract against Ethanol-Induced Hippocampal Oxidative Stress and Spatial Memory Impairment in Rats. Journal of Medicinal Plants Research, 5, 915-921.

[55]   Blokland, A., Geraerts, E. and Been, M.A. (2004) Detailed Analysis of Rat’s Spatial Memory in a Probe Trial of a Morris Task. Behavioural Brain Research, 154, 71-75.

[56]   Das, A., Dikshit, M. and Nath, C. (2005) Role of Molecular Isoforms of Acetylcholinesterase in Learning and Memory Functions. Pharmacology Biochemistry & Behavior, 81, 89-99.

[57]   Chance, B. and Maehly, A.C. (1955) Assay of Catalase and Peroxidases. Methods in Enzymology, 11, 764-775.

[58]   Kakkar, P., Das, B. and Viswanathan, P.N.A. (1984) Modified Spectrophotometric Assay of Superoxide Dismutase. Indian Journal of Biochemistry and Biophysics, 21, 130-132.

[59]   Carlberg, I. and Mannervik, E.B. (1975) Glutathione Level in Rat Brain. Journal of Biological Chemistry, 250, 4475- 4480.

[60]   Habig, W.H., Pabst, M.J. and Jakoby, W.B. (1974) Glutathione-S-Transferases: The First Enzymatic Step in Mercapturic Acid Formation. Journal of Biological Chemistry, 249, 7130-7139.

[61]   Jollow, D.J., Mitchell, J.R., Zampaglione, N. and Gillete, J.R. (1974) Bromobenzene Induced Liver Necrosis, Protective Role of Glutathione and Evidence for 3, 4-Bromobenzene Oxide as a Hepatotoxic Metabolite. Pharmacology, 11, 151-169.

[62]   Mohandas, J., Marshal, J.J., Duggin, G.G., Horvath, J.S. and Tiller, D.J. (1984) Differential Distribution of Glutathione and Glutathione-Related Enzymes in Rabbit Kidney: Possible Implications in Analgesic Nephropathy. Biochemical Pharmacology, 33, 1801-1807.

[63]   Iqbal, M., Sharma, M.D., Zadeh, H.R., Hasan, N., Abdulla, M. and Athar, M. (1996) Glutathione Metabolizing Enzymes and Oxidative Stress in Ferric Nitrilotriacetate (Fe-NTA) Mediated Hepatic Injury. Redox Report, 2, 385-391.

[64]   Ellman, G.L., Courtney, K.D., Andres, V. and Featherstone, R.M. (1996) A New and Rapid Colorimetric Determination of Acetylcholinesterase Activity. Biochemical Pharmacology, 7, 88-95.

[65]   Kumar, G.P. and Khanum F. (2012) Neuroprotective Potential of Phytochemicals. Pharmacognosy Reviews, 6, 81-90.

[66]   Sattayasai, J., Chaonapan, P., Arkaravichie, T., Soi-ampornkul, R., Junnu, S., Charoensilp, P., et al. (2013) Protective Effects of Mangosteen Extract on H2O2-Induced Cytotoxicity in SK-N-SH Cells And Scopolamine-Induced Memory Impairment in Mice. PLoS ONE, 8, e85053.

[67]   Swaroop, T.V.S.S., Banerjee, S. and Handral, M. (2014) Neuroprotective Evaluation of Leaf Extract of Dalbergia sissoo in 3-Nitropropionic Acid Induced Neurotoxicity in Rats. International Journal of Pharmaceutical Sciences and Drug Research, 6, 41-47.

[68]   Sahoo, H.B., Mandal, P.K., Bhattamisra, S.K., Bhaiji, A. and Sagar, R. (2014) A New Weapon for Memory Power: Elephantopus scaber (Linn.). International Journal of Nutrition, Pharmacology, Neurological Diseases, 4, 64-67.

[69]   Wang, J., Wang, X., Lv, B., Yuan, W., Feng, Z. and Weidong, M.I. (2014) Effects of Fructus akebiae on Learning and Memory Impairment in a Scopolamine-Induced Animal Model of Dementia. Experimental and Therapeutic Medicine, 8, 671-675.

[70]   Celik, V.K., Ersan, E., Ersan, S., Bakir, S. and Dogan, O. (2013) Plasma Catalase, Glutathione-S-Transferase and Total Antioxidant Activity Levels of Children with Attention Deficit and Hyperactivity Disorder. Advances in Bioscience and Biotechnology, 4, 183-187.

[71]   Ansari, M.A. and Scheff, S.W. (2010) Oxidative Stress in the Progression of Alzheimer Disease in the Frontal Cortex. Journal of Neuropathology & Experimental Neurology, 69, 155-167.

[72]   Fang, Y.Z., Yang, S. and Wu, G. (2000) Free Radicals, Antioxidants, and Nutrition. Nutrition. 18, 872-879.

[73]   Hayes, J.D., Flanagan, J.U. and Jowsey, I.R. (2005) Glutathione Transferases. Annual Review of Pharmacology and Toxicology, 45, 51-88.

[74]   Yadav, P., Sarkar, S. and Bhatnagar, D. (1997) Action of Capparis deciduas against Alloxan-Induced Oxidative Stress and Diabetes in Rat Tissues. Pharmacological Research, 36, 221-228.

[75]   Maritim, A.C., Sanders, R.A. and Watkins, J.B. (2003) Effects of α-Lipoic Acid on Biomarkers of Oxidative Stress in Streptozotocin-Induced Diabetic Rats. Journal of Nutritional Biochemistry, 14, 288-294.

[76]   Sultana, R., Perluigi, M. and Butterfield, D.A. (2013) Lipid Peroxidation Triggers Neurodegeneration: A Redox Proteomics View into the Alzheimer Disease Brain. Free Radical Biology and Medicine, 62, 157-169.

[77]   Lauderback, C.M., Hackett, J.M., Huang, F.F., Keller, J.N., Szweda, L.I., Markesbery, W.R., et al. (2001) The Glial Glutamate Transporter, GLT-1, Is Oxidatively Modified by 4-Hydroxy-2-Nonenal in the Alzheimer’s Disease Brain: The Role of Abeta1-42. Journal of Neurochemistry, 78, 413-416.

[78]   Khan, R.A. (2012) Effects of Launaea procumbens on Brain Antioxidant Enzymes and Cognitive Performance of Rat. BMC Complementary and Alternative Medicine, 12, 219.

[79]   Mughal, A. (2010) Acetylcholine and SK Channels Involved in Learning and Memory.

[80]   Uddin, M.S., Nasrullah, M., Hossain, M.S., Rahman, M.M., Sarwar, M.S., Amran, M.S., et al. (2016) Evaluation of Nootropic Activity of Persicaria flaccida on Cognitive Performance, Brain Antioxidant Markers and Acetylcholinesterase Activity in Rats: Implication for the Management of Alzheimer’s Disease. American Journal of Psychiatry and Neuroscience, 4, 26-37.