Neuroprotective Effects of Asparagus racemosus Willd and Withania Somnifera Dunal: An In Vitro And In Vivo Study

Authors

  • Durgesh Sharma  Department of Zoology, M.P. Government P.G. College, Chittorgarh, Rajasthan, India

Keywords:

A.racemosus, Antioxidants, Hippocampal cell culture, Neuroprotection, W.somnifera

Abstract

Aims of the study   was to investigate and compare  the  protective effects of  two plants Asparagus racemosus  Willd  (AR) and Withania somnifera Dunal (WS) in  in vitro and in vivo  models of neurodegeneration.  Further, it was also tested that the protective action was due to antioxidant activity of both the plants.  In vitro  study was carried out in hippocampal neuron cells in primary culture  and in  vivo in young adult  Wistar rats.  Root   extracts  of A.racemosus and  W.somnifera  were obtained from a  authorized supplier.  To asses  neuroprotective  action  of  the plants,  cell viability tests ( AO/EB staining, MTT assay, LDH leakage assay), extracellular Free Ca2+,  ROS determination, GSH-GSSG ratio  and  lipid peroxidation were studied in  in vitro.  In  in vivo  study,  ROS determination,  Free Ca2+,  GSH-GSSG ratio, GPx level and lipid peroxidation were determined in hippocampal tissue.  In addition dose response and exposure time response was also studied in in vitro.  Results suggested that in culture, significant decrease in dead cells was observed after   treatment with WS as compared to   AR+WS.  MTT assay also confirmed these observations. A significant decrease in Free Ca2+ ,   ROS and lipid peroxidation and increase in GSH-GSSG ratio was observed.  In  hippocampal tissue as well ,  a significantly decrease in  ROS level,  Free Ca2+,  GPx level and lipid peroxidation  but increase in GSH-GSSG ratio  was observed.  Study thus demonstrates that AR and WS both have potent neuroprotective action and that   this effect is due to their antioxidant properties

References

  1. Halliwel B. Oxidative stress and neurodegeneration. Where are we now? J Neurochem 2006; 97:1634-1658.
  2. Beal MF. Aging, energy and oxidative stress in neurodegenerative diseases, Ann Neurol 2004; 38: 357-366
  3. Liu Q, Xie F, Rolson R, Moreira PI, Nunomura A, Zhu X, Smith MA, Perry G. Prevention and treatment of Alzheimer’s disease and aging: antioxidants, Mini Rev Med Chem 2007;7 : 171-180.
  4. Anaconda TS, Reddy PH. Can herbs provide a new generation of drugs for treating Alzheimer’s disease? Brain Res 2005; 50 : 361-376.
  5. Dos-santos –Neto LL, de Vilhena Toledo MA, Medeiros-souza P, de Souza GA. The use of herbal medicine in Alzheimer’s disease- a systematic review, Evi based Compl Alt Med 2006; 3: 441-445.
  6. Jing B, Jing Bo, Liu HJ, Lai C, Zhang, XL, An Li-J. Protective effects of Catalpol against H2O2 induced oxidative stress in astrocyte primary culture, Neurosci Lett 2008; 442: 224­-227
  7. Limpeanchob N, Jaipan S, Rattanakaruna S, Phompattayarat W, Ingkanian K. Neuroprotective effects of Baccopa monnieri on beta amyloid induced cell death. J Ethanopharmaco 2008; 120: 112-117.
  8. Tateno M, Kai W, Ono T, Saito S, Hashimoto E, Saito T. Neuroprotective effects of Yi-gen-san against beta amyloid -induced toxicity in cortical neurons, Prog Neuropsychopharmacoland Biol Psy 2009: 32: 1704-1707.
  9. Bastianetto S, Ramassamy C, Dore S, Christen Y, Poirier J, Quirion R. The Ginkgo biloba extract (EGb 761) protects hippocampal neurons against cell death induced by beta-amyloid, Eur J Neurosci 2000; 12: 1882–1890.
  10. Bhatnagar M, Shukla SD, Jain S, Mundra A. Cytoprotective effects of Shankpushpi and E.alsenoides preparation on hippocampal cells in mice. Indian Drugs 2000; 37: 280-285
  11. Shukla, SD, Jain S, Sharma K, Bhatnagar M. Stress induced neurodegenration and protective effects on Semecarpus anacardium Linn: and Withania somnifera Dunn inhippocampus of albino mice: an ultrastructural study, Ind J Exp Biol 2000; 38: 1007-1013.
  12. Jain S, Shukla SD, Sharma K, Bhatnagar M. Neuroprotective effects of Withania somnifera Dunal in hippocampal sub regions of female rats, Phytother Res 2001; 15: 1-5.
  13. Sankar SR, Manivassagam T, Surendran S. Ashwagandha leaf extract: a potential agent in treating oxidative damage and physiological abnormalities seen in mouse model of Parkinson’s disease, Neurosci Lett 2009; 45: 411-419.
  14. Weggenn S, Rogers M, Erikson J. NSAIDS ; small molecules for prevention of Alzheimer’s disease or precursors for drug development, Trends Pharm Sci 2007; 28: 536-543.
  15. Soloman, B. Antibody-mediated immunotherapy for Alzhemer’s disease, Curr Opin Investig drugs 2007; 8: 519-524.
  16. Sharma PV, Sodasangahrdayam- essential of Ayurvveda. Motilal Banarsidas Publishers Pvt Ltd Delhi(1993).
  17. Puri HS, Rasayana- ayurvedic herbs for longivity and rejuvenation. Taylor and Francis, London (2003).
  18. Saxena VK, Chourasia S. A new isoflavone from the roots of Asparagus racemosus, Fitotherapia 2001; 72: 307-309.
  19. Wiboonpun N, Phuwapraisirissan P. Tip-pyang S, Identification of antioxidant compound from Asparagus racemosus, Phytother res2004; 18: 771-773.
  1. Parihar MS, Hemnani T. Experimental excitootoxicity provokes oxidative damage in mice brain and attenuation by extract of Asparagus racemosus, J Neurotransm 2004; 111: 1-2
  2. Weiner MA, Weiner J, Ashwagandha ( Indian ginseng) In: Herbs that Heal. Quantum Books. Mill Valey , CA, (1994) pp70-72. 
  3. Gupta GL, Rana AC, Withania somnifera( aswagandha ) : A review. Pharmacog Rev 2007; 1: 129-135.
  4. Bhattacharya S K, Bhattacharya A, Chakrabarhti A, Adaptogenic activity of Siotone, a polyherbal formulation of Ayurvedic rasayanas, Ind J Exp Biol 2000; 38: 119-128.
  5. Bhattacharya SK, Murogannandam AV, Adaptogenic activity of Withania somnifera : An experimental study using a rat model of chronic stress. Pharmacol Biochem Beh 2003; 75: 547-555.
  6. Ahmed M, Saleem S, Ahmed AS, Ansari MA, Yousuf S, Hoda MN, Islam F. Neuroprotective effects of Withania somnifera root on 6-hydroxydopamine induced Parkinsonism in rats, Human. Exp Toxicol 2005; 24: 137-147.
  7. Bhattacharya SK, Satyan KS, Chakrabarti A. Effect of Tarsina . an yurvedic herbal formulation on pancreatic formulation islet superoxide dismutase activity in hyperglycemic rats, Ind J Exp Biol 1997; 35: 297-299.
  8. Schelibs R, Liebmann SK, Bhattacharya SK, Kumar A, Ghosal S, Bigl V. Systemic administration of defind extracts of Withania somnifera ( Indian ginseng ) and shilajit differentially affects cholinergic but not glutamatergic and GABAergic markers in rat brain, Neurochem Int 1997; 302: 181-190.
  9. Bhatnagar M, Sharma D, Salvi M. Neuroprotective Effects of Withania somnifera Dunal.: A Possible Mechanism. Neurochem Res , 2009;
  10. Sunanda Rao BS, Raju TR. Corticosterone attenuates zinc-induced neurotoxicity in primary hippocampal cultures, Brain Res 1988; 791: 295-298.
  11. Spector DL, Goldman RD, Leinwand L A, Cells –A Laboratory Manual;Volume-1: Culture and Biochemical Analysis of Cells. Cold Spring Harbor Laboratory Press , 1999.
  12. Hansen MB, Nielsen SE, Berg K, Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J Imm Meth 1989 ;119: 203-210.
  13. Myhre O, Andersen JA, Aarnes H, Fonnum F. Evaluation of the probes 2’,7’­dichlorofluorescin diacetate , luminol and lucigenin as indicators of reactive species formation. Biochem Pharmacol 2003: 65 9: 1575-1582.
  14. Mattson MP, Tomaselli KJ, Rydel RE. Calcium destabilizing and neurodegenerative effects of aggregated beta-amyloid peptide are attenuated by basic FGF. Brain Res 1993; 621: 35-49.
  15. Koh JY, Choi DW. Quantitative determination of glutamate mediated cortical neuronal injury in cell culture by lactate dehydrogenase efflux assay. J Neurosci Meth 1987; 20: 83-90
  16. Hissin PJ, Hilf RA. Fluorimetric method for determination of oxidized and reduced glutathione in tissue, Anal Biochem 1976; 74: 214-226.
  17. Utley HG, Bergheim F, Hochstein P. Effects of sulfa hydral reagents on peroxidation of microsomes, Arch Biochem Biophy 1976; 118: 29-32.
  18. Bhattacharya A, Ghosal S, Bhattacharya SK, Antioxidant effects of Withania sonmifera glycowithanolides in chronic foot shock stress induced perturbations of oxidative free radical scavenging enzymes and lipid peroxidation in rat frontal cortex and striatum. J Ethanopharmacol 2001; 74: 1-6.
  19. Lowry, OH, Rosenberg NJ, Farr AL, Randall RJ. Protein measurement with Folin-phenol reagent. J Biol Chem 1951; 193: 265-275.
  20. Le Bel CP, Ali SF, Mckee M , Bondy S C. Organometal-induced increases in oxygen reactive species : the potential of 2’7’-dicholorofluorescin diacetate as an index of neurotoxic damage. Toxicol App Pharmacol 1999; 104: 17-24.
  21. Buege JA, Aust SD. The thiobarbituric acid assay. Methods in Enzymology 1978; 52: 306.
  22. Parihar MS, Hemnani T. Phenolic antioxidants attenuate hippocampal neuronal cell damage against kainic acid induced excitotoxicity, J Biosci 2003; 28: 121-128.
  23. Meister A, Selective modification of glutathione metabolism. Science 1983; 220 : 472-477.
  24. Cardozo-Pelaez F, Brooks PJ, Stedeford T, Song S, Sanchez-Ramos J. DNA damage repair and antioxidant system in brain regions: A correlative study. Free Rad Biol Med 2000; 28: 779-785.
  25. Tirmenstein MA, Nichols-Grzemski FA, Zhang JG, Fariss MW. Glutathione depletion and production of reactive oxygen species in isolated hepatocytes suspension. Chem Biol Intern 2000; 127: 201-217.
  26. Olney JW. Brain lesions, obesity and other disturbances in mice treated with monosodium glutamate. Science 1969; 164: 719-721.
  27. Michaels RL, Rothman SM. Glutamate neurotoxicity in in vitro antagonist pharmacology and intracellular calcium concentrations. J Neurosci 1990; 10: 283-292.
  28. Lowy MT, Gault L, Yamamoto BK. Adrenelectomy attenuates stress induced elevation in extracellular glutamate concentration in hippocampus. J Neurochem 1993; 61(1993) 1957-1960.
  29. Calabrese V, Mancuso C, Calvani M, Rizzarelli E, Butterfield DA , Stell AMG. Nitric oxide in the central nervous system: neuroprotection versus neurotoxicity. Nat Rev Neuroscience 2007; 8: 766-775.
  30. Kamat JP, Boloor KK, Devasagayam TP, Vnkatachalam SR, Antioxidant properties of Asparagus racemosus against damage induced by gamma radiation in rat liver mitochondria. J Ethanopharmacol, 2000; 71: 425-435.
  31. Gupt SK, Dua A, Vohra BP. Withania somnifera (Ashwagandha ) attenuates antioxidant defense in aged spinal cord and inhibits copper induced lipid peroxidation and protein oxidative modification. Drug Metabol Intern 2003:19: 211-222.

International Journal of Scientific Research in Science, Engineering and Technology

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Published

2014-04-30

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Volume 1, Issue 1, -2014

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Research Articles

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[1]
Durgesh Sharma, " Neuroprotective Effects of Asparagus racemosus Willd and Withania Somnifera Dunal: An In Vitro And In Vivo Study, International Journal of Scientific Research in Science, Engineering and Technology(IJSRSET), Print ISSN : 2395-1990, Online ISSN : 2394-4099, Volume 1, Issue 1, pp.111-121, -2014.