Neuroprotective effects of Withania coagulans root extract on CA1 hippocampus following cerebral ischemia in rats

Document Type: Original Research Article

Authors

1 Department of Anatomy, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran

2 Infectious Diseases and Tropical Medicine Research Center, and Department of histology, School of Medicine, Zahedan University of Medical sciences, Zahedan, Iran

3 Department of Histology, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran

4 Department of Medicinal and Aromatic Plant, High Complex Education of Saravan, Saravan, Iran

Abstract


Objective: Oxygen free radicals may be implicated in the pathogenesis of ischemia reperfusion damage. The beneficial effects of antioxidant nutrients, as well as complex plant extracts, on cerebral ischemia-reperfusion injuries are well known. This study was conducted to determine the effects of the hydro-alcoholic root extract of Withania coagulans on CA1 hippocampus oxidative damages following global cerebral ischemia/reperfusion in rat.

 

Materials and Methods: Male Wistar rats were randomly divided in five groups: control, sham operated, Ischemia/ Reperfiusion (IR), and Withania Coagulans Extract (WCE) 500 and 1000mg/kg + I/R groups.  Ischemia was induced by ligation of bilateral common carotid arteries for 30 min after 30 days of WCE administration. Three days after, the animals were sacrificed, their brains were fixed for histological analysis (NISSL and TUNEL staining) and some samples were prepared for measurement of malondialdehyde (MDA) level and superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activity in hippocampus.
Results: WCE showed neuroprotective activity by significant decrease in MDA level and increase in the SOD, CAT and GPx activity in pretreated groups as compared to I/R groups (p<0.001).  The number of intact neurons was increased while the number of TUNEL positive neurons in CA1 hippocampal region in pretreated groups were decreased as compared to I/R group (p<0.001).
Conclusion: WCE showed potent neuroprotective activity against oxidative stress-induced injuries caused by global cerebral ischemia/ reperfusion in rats probably by radical scavenging and antioxidant activities.

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Main Subjects


Aabdallah DM, Eid NI. 2004. Possible neuroprotective effects of lecithin and α-tocopherol alone or in combination against ischemia/reperfusion insult in rat brain. J Biochem Mol Toxicol, 18:273-278.

 Ahmad A, Khan MM, Raza SS, Javed H, Ashafaq M, Islam F, Safhi MM, Islam F. 2012. Ocimum sanctum attenuates oxidative damage and neurological deficits following focal cerebral ischemia/reperfusion injury in rats. Neurol Sci, 33:1239-1247.

Ajami M, Eghtesadi S, Razaz JM, Kalantari N, Habibey R, Nilforoushzadeh MA, Zarrindast M, Pazoki-Toroudi H. 2011. Expression of Bcl-2 and Bax after hippocampal ischemia in DHA+ EPA treated rats. Neurol Sci, 32:811-818.

Akyol Ö, Herken H, Uz E, Fadıllıoǧlu E, Ünal S, Söǧüt S, Özyurt H, Savaş HA. 2002. The indices of endogenous oxidative and antioxidative processes in plasma from schizophrenic patients: the possible role of oxidant/antioxidant imbalance. Prog Neuro-Psychoph, 26:995-1005.

Al-Omar FA, Nagi MN, Abdulgadir MM, Al Joni KS, Al-Majed AA. 2006. Immediate and delayed treatments with curcumin prevents forebrain ischemia-induced neuronal damage and oxidative insult in the rat hippocampus. Neurochem Res, 31:611-618.

Anderson  MF, Nilsson M, Eriksson PS, Sims NR. 2004. Glutathione monoethyl ester provides neuroprotection in a rat model of stroke. Neurosci lett, 354:163-165.

Awwad M, El-Azim SAA, Marzouk F, El-Ghany E, Barakat M. 2011. Radio-protective chelating agents against DNA oxidative damage. Afr J Biochem Res, 5:106-112.

Baitharu I, Jain V, Deep SN, Shroff S, Sahu JK, Naik PK, Ilavazhagan G. 2014. Withanolide A Prevents Neurodegeneration by Modulating Hippocampal Glutathione Biosynthesis during Hypoxia. PloS one 9.

Bast A, Barr P. 1997. The antioxidant/prooxidant balance in neurodegeneration and neuroprotection. Neuroprotection in CNS Diseases. Marcel Deccer Inc, 147-159.

Beigomi M, Mohammadifar MA, Hashemi M, Ghods rohani M, Senthil K, Valizadeh M. 2014. Biochemical and rheological characterization of a protease from fruits of Withania coagulans with a milk-clotting activity. Food Sci Biotechnol, 23: 1805-1813.

Bémeur C, Qu H, Desjardins P, Butterworth RF. 2010. IL-1 or TNF receptor gene deletion delays onset of encephalopathy and attenuates brain edema in experimental acute liver failure. Neurochem Int, 56:213-215.

Bhatnagar M, Meena P, Barbar S, Joshi C. 2013. Neuroprotective response of the hippocampus region of the brain to Withania somnifera and Asparagus racemosus root extract: An in vitro study.J Med Plants Res, 7:2259-2264.

Bin J, Wang Q, Zhuo Y-Y, Xu J-P, Zhang H-T. 2012. Piperphentonamine (PPTA) attenuated cerebral ischemia-induced memory deficits via neuroprotection associated with anti-apoptotic activity. Metab Brain Dis, 27:495-505.

Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem, 72:248-254.

Budhiraja R, Sudhir S, Garg K. 1984. Antiinflammatory activity of 3 β-Hydroxy-2, 3-dihydro-withanolide F. Planta Medica, 50:134-136.

Candelario-Jalil E, Mhadu NH, Al-Dalain SM, Martı́nez G, León OS. 2001. Time course of oxidative damage in different brain regions following transient cerebral ischemia in gerbils. Neurosci Res, 41:233-241.

Chandrashekhar V, Ranpariya V, Ganapaty S, Parashar A, Muchandi A. 2010. Neuroprotective activity of Matricaria recutita Linn against global model of ischemia in rats. J Ethnopharmacol, 127:645-651.

Chaudhary G, Sharma U, Jagannathan NR, Gupta YK. 2003. Evaluation of Withania somnifera in a middle cerebral artery occlusion model of stroke in rats. Clin Exp Pharmacolo P, 30:399-404.

Colbourne F, Sutherland GR, Auer RN. 1999. Electron microscopic evidence against apoptosis as the mechanism of neuronal death in global ischemia. J neuroscience, 19:4200-4210.

Doyle KP, Simon RP, Stenzel-Poore MP. 2008. Mechanisms of ischemic brain damage. Neuropharmacol, 55:310-318.

Floyd RA. 2002. Oxidative stress in brain aging Implications for therapeutics of neurodegenerative diseases. Neurobiol Aging, 23:795–807.

Goth L. 1991. A simple method for determination of serum catalase activity and revision of reference range. Clinica Chimica Acta, 196:143-151.

Gupta S, Sharma SS. 2006. Neuroprotective effects of trolox in global cerebral ischemia in gerbils. Biol Pharm Bull, 29:957-961.

Hemalatha S, Wahi A, Singh P, Chansouria J. 2004. Hypoglycemic activity of Withania coagulans Dunal in streptozotocin induced diabetic rats. J Ethnopharmacol, 93:261-264.

Jaiswal D, Kumar Rai P, Watal G. 2009. Antidiabetic effects of Withania coagulans in experimental rats.  Indian J Clin Biochem, 24: 88-93.

Jimenez-Jimenez F, Garcia-Ruiz P, de Bustos F. 1996. Calcium, neuronal death and neurological disease. Rev Neurologia, 24:1199-1209.

Jung HW, Mahesh R, Bae HS, Kim YH, Kang JS, Park Y-K. 2011. The antioxidant effects of Joongpoongtang 05 on brain injury after transient focal cerebral ischemia in rats. J Natl Med Assoc, 65:322-329.

Kakkar P, Das B, Viswanathan P. 1984. A modified spectrophotometric assay of superoxide dismutase. Indian J Biochem Biophys, 21:130-132.

Kong X, Kong W, Miao G, Zhao S, Chen M, Zheng X, Bai J. 2014. Pretreatment with scutellaria baicalensis stem-leaf total flavonoid protects against cerebral ischemia/reperfusion injury in hippocampal neurons. Neural Regen Res, 9:2066-2073.

Lalsare S, Chutervedi A. 2010. Anti-inflammatory and anthihyperlipidemic activity of various extract of fruits Withania coagulance. Pharmacology, 1:101-107.

Lee J, Won M. 2014. Neuroprotection of antioxidant enzymes against transient global cerebral ischemia in gerbils. Anat Cell Biol, 47:149-156.

Mathur D, Agrawal R, Shrivastava V. 2011. Phytochemical Screening and Determination of Antioxidant Potential of Fruits Extracts of Withania coagulans. Rec Res SciTech, 3:26-29.

Maurya R. 2010. Chemistry and pharmacology of Withania coagulans: an Ayurvedic remedy. J Pharm Pharmacol, 62:153-160.

Mohagheghi F, Bigdeli MR, Rasoulian B, Hashemi P, Pour MR. 2011. The neuroprotective effect of olive leaf extract is related to improved blood–brain barrier permeability and brain edema in rat with experimental focal cerebral ischemia. Phytomedicine, 18:170-175.

Mohammadi MT, Amini R, Jahanbakhsh Z, Shekarforoush S. 2013. Effects of atorvastatin on the hypertension-induced oxidative stress in the rat brain. Iran Biomed J, 17:152-157.

Mohanty I, Arya DS, Dinda A, Talwar KK, Joshi S, Gupta SK. 2004. Mechanisms of cardioprotective effect of Withania somnifera in experimentally induced myocardial infarction. Basic Clin Pharmacol, 94:184-190.

Oh JH, Kwon TK. 2009. Withaferin A inhibits tumor necrosis factor-α-induced expression of cell adhesion molecules by inactivation of Akt and NF-κB in human pulmonary epithelial cells. Int Immunopharmacol, 9:614-619.

Ohkawa H, Ohishi N, Yagi K. 1979. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem, 95:351-358.

Ozerol E, Bilgic S, Iraz M, Cigli A, Ilhan A, Akyol O. 2009. The protective effect of erdosteine on short-term global brain ischemia/reperfusion injury in rats. Prog Neuro-Psychopharmacoph, 33:20-24.

Paglia DE, Valentine WN. 1967. Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J lab Clin Medicine, 70:158-169.

Pham-Huy LA, He H, Pham-Huy C. 2008. Free radicals, antioxidants in disease and health. Int J Biomed Sci, 4:89-96.

Prasad S, Kumar R, Patel D, Hemalatha S. 2010. Wound healing activity of Withania coagulans in streptozotocin-induced diabetic rats. Pharm Biol, 48:1397-1404.

Ramana KV, Srivastava S, Singhal SS. 2013. Lipid peroxidation products in human health and

 disease. Oxid Med Cell longev. doi.org/10.1155/2013/583438

Rao KVR, Tong X, Alvarez VM, Norenberg MD. 2010. Marked potentiation of cell swelling by cytokines in ammonia-sensitized cultured astrocytes. J Neuroinflamm, 7:66-68.

Rossi DJO, T. Attwell, D. 2000. Glutamate release in severe brain ischemia is mainly by reversed uptake. Nature, 403:316-321.

Salwaan C, Singh A, Mittal A, Singh P. 2012. Investigation of the Pharmacognostical, Phytochemical and Antioxidant Studies of Plant Withania Coagulans Dunal. J Pharmacog Phytochem, 1:36-44.

Sarbishegi M, Mehraein F, Soleimani M. 2014. Antioxidant role of oleuropein on midbrain and dopaminergic neurons of substantia nigra in aged rats. Iran Biomed J, 18:16-22

Shah ZA, Gilani RA, Sharma P, Vohora SB. 2005. Cerebroprotective effect of Korean ginseng tea against global and focal models of ischemia in rats. J Ethnopharmacol, 101:299-307.

Shukla K, Dikshit P, Tyagi MK, Shukla R, Gambhir JK. 2012. Ameliorative effect of Withania coagulans on dyslipidemia and oxidative stress in nicotinamide–streptozotocin induced diabetes mellitus. Food Chem Toxicol, 50:3595-3599.

Thiyagarajan M, Sharma SS. 2004. Neuroprotective effect of curcumin in middle cerebral artery occlusion induced focal cerebral ischemia in rats. Life Sci, 74:969-985.

Vandresen-Filho S, Martins WC, Bertoldo DB, Mancini G, De Bem AF, Tasca CI. 2015. Cerebral cortex, hippocampus, striatum and cerebellum show differential susceptibility to quinolinic acid-induced oxidative stress. J Neurol Sci, DOI 10.1007/s10072-015-2180-7.

Yabuki Y, Fukunaga K. 2013. Oral administration of glutathione improves memory deficits following transient brain ischemia by reducing brain oxidative stress. Neuroscience, 250:394-407.

Yan BC, Park JH, Ahn JH, Kim IH, Park OK, Lee JC, Yoo KY, Choi JH, Lee CH, Hwang IK. 2014. Neuroprotection of posttreatment with risperidone, an atypical antipsychotic drug, in rat and gerbil models of ischemic stroke and the maintenance of antioxidants in a gerbil model of ischemic stroke. J Neurosci Res, 92:795-807.

Yigitkanli K, Pekcec A, Karatas H, Pallast S, Mandeville E, Joshi N, Smirnova N, Gazaryan I, Ratan RR, Witztum JL. 2013. Inhibition of 12/15-lipoxygenase as therapeutic strategy to treat stroke. Ann Neurol, 73:129-135.

Yoo K-Y, Lee CH, Park JH, Hwang IK, Park OK, Kwon S-H, Choi JH, Kim D-j, Kwon Y-G, Kim Y-M. 2011. Antioxidant enzymes are differently changed in experimental ischemic hippocampal CA1 region following repeated restraint stress. J Neurol Sci, 302:33-42.

Yu ZF, Bruce-Keller AJ, Goodman Y, Mattson MP. 1998. Uric acid protects neurons against excitotoxic and metabolic insults in cell culture, and against focal ischemic brain injury in vivo. J Neurosci Res, 53:613-625.