NEUROPROTECTIVE AND COGNITIVE ENHANCING EFFECT OF METHANOLIC MORUS ALBA LEAF FRACTION IN U87MG CELL LINES AND EXPERIMENTAL RAT MODEL
Objective: The present study aims to investigate the protective effect of methanol fraction of Morus alba (MEMA) leaves against hydrogen peroxide (H2O2)-induced U87MG cell toxicity and aluminum fluoride (ALF)-induced rat toxicity.
Methods: The study was divided into in vitro and in vivo sections. U87MG cell lines were pre-treated with different fractions of MEMA for 20 h and further tested against 1000 ϻM of H2O2. The best fraction from in vitro studies was used to study the protective effects against ALF-induced neurotoxicity. Rats were divided i nto five different groups, and MEMA (200 and 400 mg/kg p.o) was administered for 14 days to the animals with α-tocopherol as the standard drug treatment. Behavioral studies were assessed using Barnes maze. The major biochemical measurements included catalase, superoxide dismutase and glutathione reductase, lipid peroxidation (LPO), and acetylcholinesterase (AchE) levels.
Results: In vitro studies indicated MEMA as a potential candidate followed by AQMA and ethyl acetate. The MEMA fraction was able to ameliorate ALF-induced neurotoxicity in the behavioral assessment. The higher antioxidant content in the fraction decreased the LPO levels from 250±4.07 to 115±3.22 as well as elevated the levels of most of the endogenous antioxidant enzyme levels. AchE levels were also decreased to 33.89±0.71 from 38.94±0.64.
Conclusion: Although the results obtained indicate that MEMA could significantly suppress oxidative stress-induced central neuronal damage both in vitro and in vivo, further mechanistic studies are required to delineate its neuroprotective pathway.
2. Ghezzi L, Scarpini E, Galimberti D. Disease-modifying drugs in Alzheimer’s disease. Drug Des Devel Ther 2013;7:1471-8.
3. Chiu PY, Tsai CT, Chen PK, Chen WJ, Lai TJ. Neuropsychiatric symptoms in Parkinson’s disease dementia are more similar to Alzheimer’s disease than dementia with lewy bodies: A Case-control study. PLoS One 2016;11:e0153989.
4. Sejvar JJ. Global burden of neurological disease: What’s in a name? Lancet Neurol 2017;16:858-9.
5. Winblad B, Amouyel P, Andrieu S, Ballard C, Brayne C, Brodaty H, et al. Defeating Alzheimer’s disease and other dementias: A priority for European science and society. Lancet Neurol 2016;15:455-532.
6. Purushothuman S. Oxidative stress in neurodegenerative conditions and the protective potential of a natural antioxidant, dietary saffron. Oxid Antioxid Med Sci 2015;4:112-3.
7. Wimo A, Guerchet M, Ali GC, Wu YT, Prina AM, Winblad B, et al. The worldwide costs of dementia 2015 and comparisons with 2010. Alzheimers Dement 2017;13:1-7.
8. Mansour SZ, Moustafa EM, Hassan AA, Thabet NM. Protective role of krill oil against estrogen deficiency induced neurodegeneration in ovariectomized mice. Indian J Exp Biol 2017;55:279-80.
9. Betteridge DJ. What is oxidative stress? Metabolism 2000;49:3-8.
10. Chonpathompikunlert P, Han J, Toh K, Isoda H, Nagasaki Y. TEMPOL protects human neuroblastoma SH-SY5Y cells against ?-amyloid-induced cell toxicity. Eur J Pharmacol 2011;650:544-9.
11. Milton NG. Role of hydrogen peroxide in the aetiology of Alzheimer’s disease: Implications for treatment. Drugs Aging 2004;21:81-100.
12. Singh JC, Alagarsamy V, Parthiban P, Selvakumar P, Reddy YN. Neuroprotective potential of ethanolic extract of Pseudarthria viscida (L) Wight and arn against beta-amyloid (25-35)-induced amnesia in mice. Indian J Biochem Biophys 2011;48:197-201.
13. Mclachlan D. Aluminum and Alzheimer’s disease. Alzheimer Dis Assoc Disord 1987;1:209-10.
14. Imran M, Khan H, Shah M, Khan R, Khan F. Chemical composition and antioxidant activity of certain Morus species. J Zhejiang Univ Sci B 2010;11:973-80.
15. Raj A, Mruthunjaya K, Madhunapantula S, Manjula S. Comparative assessment of the anti-oxidant and anti-clastogenic activity of Morus alba leaves. Free Radic Antioxid 2016;7:123-7.
16. Kumar VS. Studies to Assess the Effect of Morus alba Against Scopolamine Induced Memory Impairment and Amyloid ?-Peptide1-40 Induced Neurotoxicity. Master of Pharmacy Thesis. Mysore: Jagadguru Sri Shivarathreeshwara University; 2014.
17. Thakor V, Poddar M, Dey S, Manjula S, Madhunapantula S, Pawara R, et al. Exploring the anti-breast cancer potential of flavonoid analogs. RSC Adv 2016;6:79166-79.
18. Armstrong JS, Steinauer KK, Hornung B, Irish JM, Lecane P, Birrell GW, et al. Role of glutathione depletion and reactive oxygen species generation in apoptotic signaling in a human B lymphoma cell line. Cell Death Differ 2002;9:252-63.
19. Liu K, Liu PC, Liu R, Wu X. Dual AO/EB staining to detect apoptosis in osteosarcoma cells compared with flow cytometry. Med Sci Monit Basic Res 2015;21:15-20.
20. Darcet F, Mendez-David I, Tritschler L, Gardier AM, Guilloux JP, David DJ, et al. Learning and memory impairments in a neuroendocrine mouse model of anxiety/depression. Front Behav Neurosci 2014;8:136.
21. Khan RA, Khan MR, Sahreen S. Brain antioxidant markers, cognitive performance and acetylcholinesterase activity of rats: Efficiency of Sonchus asper. Behav Brain Funct 2012;8:21.
22. Sinha AK. Colorimetric assay of catalase. Anal Biochem 1972;47:389 94.
23. Kinattingal N, Manjula SN, Mruthunjaya K, Mahalakshmi AM. Cardioprotective effect of Tamarindus indica linn against isoproterenol induced myocardial infarction in rats. Int J Pharm Pharm Sci 2016;8:254-60.
24. Babu SM, Swain S, Renuka K. Neuroprotective activity of fractional flower extracts of Mirabilis jalapa against aluminium hydrochloride induced neurotoxicity in male Wister rats. Int J Pharm Pharm Sci 2017;9:216-21.
25. Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979;95:351-8.
26. Sumithra M, Arunachalam G, Chitra V, Gowri K. Neuroprotective effect of Sargassum ilicifolium Turner Carl Agardh on acetylcholinesterase activity and attenuation of scopolamine-induced amnesia in rodents. Asian J Pharm Clin Res 2016;9:93-6.
27. Li J, Wuliji O, Li W, Jiang ZG, Ghanbari HA. Oxidative stress and neurodegenerative disorders. Int J Mol Sci 2013;14:24438-75.
28. Mushtaq N, Schmatz R, Pereira LB, Ahmad M, Stefanello N, Vieira JM, et al. Rosmarinic acid prevents lipid peroxidation and increase in acetylcholinesterase activity in brain of streptozotocin-induced diabetic rats. Cell Biochem Funct 2014;32:287-93.
29. Garcia YJ, Rodríguez-Malaver AJ, Peñaloza N. Lipid peroxidation measurement by thiobarbituric acid assay in rat cerebellar slices. J Neurosci Methods 2005;144:127-35.
30. Singh B, Pandey S, Verma R, Ansari JA, Mahdi AA. Comparative evaluation of extract of Bacopa monnieri and Mucuna pruriens as neuroprotectant in MPTP model of Parkinson’s disease. Indian J Exp Biol 2016;54:758-66.
31. Rahman K. Studies on free radicals, antioxidants, and co-factors. Clin Interv Aging 2007;2:219-36.
32. Niidome T, Takahashi K, Goto Y, Goh S, Tanaka N, Kamei K, et al. Mulberry leaf extract prevents amyloid beta-peptide fibril formation and neurotoxicity. Neuroreport 2007;18:813-6.
33. Behl C, Davis JB, Lesley R, Schubert D. Hydrogen peroxide mediates amyloid beta protein toxicity. Cell 1994;77:817-27.
34. Whittemore ER, Loo DT, Watt JA, Cotman CW. A detailed analysis of hydrogen peroxide-induced cell death in primary neuronal culture. Neuroscience 1995;67:921-32.
35. Baylock RL. Excitotoxicity: A possible central mechanism in fluoride neurotoxicity. Flouride 2007;37:301-14.
36. Barad M, Bourtchouladze R, Winder DG, Golan H, Kandel E. Rolipram, a type IV-specific phosphodiesterase inhibitor, facilitates the establishment of long-lasting long-term potentiation and improves memory. Proc Natl Acad Sci U S A 1998;95:15020-5.
37. Nade VS, Kawale LA. Targetting serotonergic pathway for anti-amnesic activity by Morus alba L. Int J Pharm Sci Drug Res 2015;7:27-32.A
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