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Neuroprotective effect of chlorogenic acid in global cerebral ischemia-reperfusion rat model

  • Gaurav Kumar
  • Sumedha Mukherjee
  • Pankaj Paliwal
  • Saumitra Sen Singh
  • Hareram Birla
  • Surya Pratap Singh
  • Sairam Krishnamurthy
  • Ranjana PatnaikEmail author
Original Article

Abstract

The ischemic cascade is initiated in the hypoperfused region of the brain that leads to neuronal cell death. Identification of multi-target inhibitor against prominent molecular mediators of ischemic cascade might be a suitable strategy to combat cerebral ischemic stroke. The present study is designed to evaluate the neuroprotective efficacy of chlorogenic acid (CGA) in the global cerebral ischemic rat model. The effective dose of CGA was evaluated on the basis of reduction in cerebral infarction area percentage, Evans blue extravasation, and restoration of brain water content. The expression of tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), and caspase-3 was evaluated by immunohistochemistry and morphological and cellular alterations in the cortex were observed by brain histology. The level of glutamate, calcium, and nitrate in different regions of the brain, as well as cerebrospinal fluid (CSF), was evaluated. The level of calcium and nitrate was compared with ifenprodil—an antagonist of N-methyl-D-aspartate receptor (NMDAR) and 7-nitroindazole—an inhibitor of neuronal nitric oxide synthase (nNOS) respectively. Further, molecular docking was performed to compare the inhibition potential of CGA against NMDAR and nNOS with their inhibitors. Dose optimization results revealed that intranasal administration of CGA (10 mg/kg b.w.) significantly reduced the cerebral infarction area, Evans blue extravasation and restored the brain water content compared with ischemia group. It also significantly reduced the calcium, nitrate, and glutamate levels compared with ischemia group in the cortex, hippocampus cerebellum, and CSF. Immunohistochemical analysis revealed that CGA significantly reduced the expression of TNF-α, iNOS, and caspase-3 as compared with the ischemia group. In molecular docking study, CGA displayed similar binding interaction as that of Ifenprodil and 7-nitroindazole with NMDAR and nNOS respectively. The current findings suggest that the treatment with CGA confers neuroprotection in global ischemic insult by inhibiting and downregulating the different molecular markers of cerebral ischemia.

Keywords

Chlorogenic acid Ischemic stroke Intranasal Immunohistochemistry Molecular docking Neuroprotection 

Notes

Acknowledgments

The authors are thankful to the Coordinator, ISLS, Banaras Hindu University (BHU) for providing the HPLC-UV facility and Chandra Prakash Patel for his help in the fluorescence microscopy. The authors are also thankful to Coordinator, School of Biomedical Engineering, Indian Institute of Technology (BHU), Varanasi for providing necessary facilities to conduct the research work.

Author contributions

GK designed and performed the experiments and co-wrote the manuscript. SM performed sample processing and co-wrote the manuscript. PP and SK performed the histology study. SSS, HB, and SPS performed the immunohistochemistry study. GK and PP statistically analyzed the study. RP conceived and supervised the complete study.

References

  1. Ahn EH, Kim DW, Shin MJ, Kwon SW, Kim YN, Kim DS, Lim SS, Kim J, Park J, Eum WS, Hwang HS, Choi SY (2011) Chlorogenic acid improves neuroprotective effect of PEP-1-ribosomal protein S3 against ischemic insult. Exp Neurobiol 20:169–175.  https://doi.org/10.5607/en.2011.20.4.169 CrossRefGoogle Scholar
  2. Amantea D, Tassorelli C, Russo R, Petrelli F, Morrone LA, Bagetta G, Corasaniti MT (2011) Neuroprotection by leptin in a rat model of permanent cerebral ischemia: effects on STAT3 phosphorylation in discrete cells of the brain. Cell Death Dis 2:e238.  https://doi.org/10.1038/cddis.2011.125 CrossRefGoogle Scholar
  3. Bouayed J, Rammal H, Dicko A, Younos C, Soulimani R (2007) Chlorogenic acid, a polyphenol from Prunus domestica (Mirabelle), with coupled anxiolytic and antioxidant effects. J Neurol Sci 262:77–84.  https://doi.org/10.1016/j.jns.2007.06.028 CrossRefGoogle Scholar
  4. Cardiff RD, Miller CH, Munn RJ (2014) Manual hematoxylin and eosin staining of mouse tissue sections. Cold Spring Harb Protoc 2014.  https://doi.org/10.1101/pdb.prot073411
  5. Cropley V, Croft R, Silber B, Neale C, Scholey A, Stough C, Schmitt J (2012) Does coffee enriched with chlorogenic acids improve mood and cognition after acute administration in healthy elderly? A pilot study. Psychopharmacology 219:737–749.  https://doi.org/10.1007/s00213-011-2395-0 CrossRefGoogle Scholar
  6. Dhawan J, Benveniste H, Luo Z, Nawrocky M, Smith SD, Biegon A (2011) A new look at glutamate and ischemia: NMDA agonist improves long-term functional outcome in a rat model of stroke. Future Neurol 6:823–834.  https://doi.org/10.2217/fnl.11.55 CrossRefGoogle Scholar
  7. Dirnagl U, Iadecola C, Moskowitz MA (1999) Pathobiology of ischaemic stroke: an integrated view. Trends Neurosci 22:391–397CrossRefGoogle Scholar
  8. Eliasson MJ, Huang Z, Ferrante RJ et al (1999) Neuronal nitric oxide synthase activation and peroxynitrite formation in ischemic stroke linked to neural damage. J Neurosci 51:2131–2144.  https://doi.org/10.4319/lo.2006.51.5.2131 Google Scholar
  9. Farbiszewski R, Bielawski K, Bielawska A, Sobaniec W (1995) Spermine protects in vivo the antioxidant enzymes in transiently hypoperfused rat brain. Acta Neurobiol Exp WarsGoogle Scholar
  10. Favie LMA, Cox AR, van den Hoogen A et al (2018) Nitric oxide synthase inhibition as a neuroprotective strategy following hypoxic-ischemic encephalopathy: evidence from animal studies. Front Neurol 9.  https://doi.org/10.3389/fneur.2018.00258
  11. Ferrer I, Friguls B, Dalfót E et al (2003) Caspase-dependent and caspase-independent signalling of apoptosis in the penumbra following middle cerebral artery occlusion in the adult rat. Neuropathol Appl Neurobiol 29:472–481.  https://doi.org/10.1046/j.1365-2990.2003.00485.x CrossRefGoogle Scholar
  12. Gagliardi RJ (2000) Neuroprotection, excitotoxicity and NMDA antagonists. Arq Neuropsiquiatr 58:583–588.  https://doi.org/10.1590/S0004-282X2000000300030 CrossRefGoogle Scholar
  13. Gorbatyuk OS, Li S, Sullivan LF, Chen W, Kondrikova G, Manfredsson FP, Mandel RJ, Muzyczka N (2008) The phosphorylation state of Ser-129 in human alpha-synuclein determines neurodegeneration in a rat model of Parkinson disease. Proc Natl Acad Sci U S A 105:763–768.  https://doi.org/10.1073/pnas.0711053105 CrossRefGoogle Scholar
  14. Graff CL, Pollack GM (2005) Nasal drug administration: potential for targeted central nervous system delivery. J Pharm Sci 94:1187–1195CrossRefGoogle Scholar
  15. Han J, Miyamae Y, Shigemori H, Isoda H (2010) Neuroprotective effect of 3,5-di-O-caffeoylquinic acid on SH-SY5Y cells and senescence-accelerated-prone mice 8 through the up-regulation of phosphoglycerate kinase-1. Neuroscience. 169:1039–1045.  https://doi.org/10.1016/j.neuroscience.2010.05.049 CrossRefGoogle Scholar
  16. Heitman E, Ingram DK (2016) Cognitive and neuroprotective effects of chlorogenic acid. Nutr Neurosci 20:32–39.  https://doi.org/10.1179/1476830514Y.0000000146 CrossRefGoogle Scholar
  17. Hu H, Sun XO, Tian F et al (2016) Neuroprotective effects of isosteviol sodium injection on acute focal cerebral ischemia in rats. Oxidative Med Cell Longev 2016:1–10.  https://doi.org/10.1155/2016/1379162 Google Scholar
  18. Hwang SJ, Kim Y-W, Park Y, Lee HJ, Kim KW (2014) Anti-inflammatory effects of chlorogenic acid in lipopolysaccharide-stimulated RAW 264.7 cells. Inflamm Res 63:81–90.  https://doi.org/10.1007/s00011-013-0674-4 CrossRefGoogle Scholar
  19. Iadecola C, Zhang F, Xu X (1995) Inhibition of inducible nitric oxide synthase ameliorates cerebral ischemic damage. Am J PhysGoogle Scholar
  20. Ichiki H, Kuroiwa T, Taniguchi I, Okeda R (1998) Delayed recovery of cortical auditory evoked potentials and blood flow precede cortical neuronal death after transient cerebral ischemia in gerbils. J Med Dent SciGoogle Scholar
  21. Ito H, Sun X-L, Watanabe M et al (2008) Chlorogenic acid and its metabolite m -coumaric acid evoke neurite outgrowth in hippocampal neuronal cells. Biosci Biotechnol Biochem 72:885–888.  https://doi.org/10.1271/bbb.70670 CrossRefGoogle Scholar
  22. Iwasaki Y, Ito S, Suzuki M, Nagahori T, Yamamoto T, Konno H (1989) Forebrain ischemia induced by temporary bilateral common carotid occlusion in normotensive rats. J Neurol Sci 90:155–165.  https://doi.org/10.1016/0022-510X(89)90098-1 CrossRefGoogle Scholar
  23. Johnston KL, Clifford MN, Morgan LM (2003) Coffee acutely modifies gastrointestinal hormone secretion and glucose tolerance in humans: glycemic effects of chlorogenic acid and caffeine. Am J Clin Nutr 47:1453–1459.  https://doi.org/10.1021/jf980737w Google Scholar
  24. Jovanović MĆ, Djukic M, Vasiljević I et al (2007) Determination of nitrate by the IE-HPLC-UV method in the brain tissues of Wistar rats poisoned with paraquat. J Serbian Chem Soc 72:347–356.  https://doi.org/10.2298/JSC0704347C CrossRefGoogle Scholar
  25. Karakas E, Simorowski N, Furukawa H (2011) Subunit arrangement and Phenylethanolamine binding in GluN1/GluN2B NMDA receptors. Nature 475:249–253.  https://doi.org/10.1038/nature10180 CrossRefGoogle Scholar
  26. Keep RF, Hua Y, Xi G (2012) Brain water content: a misunderstood measurement? Transl Stroke Res 3:263–265.  https://doi.org/10.1007/s12975-012-0152-2 CrossRefGoogle Scholar
  27. Kumar G, Patnaik R (2016) Exploring neuroprotective potential of Withania somnifera phytochemicals by inhibition of GluN2B-containing NMDA receptors: an in silico study. Med Hypotheses 92:35–43.  https://doi.org/10.1016/j.mehy.2016.04.034 CrossRefGoogle Scholar
  28. Kumar G, Patnaik R (2017) Inhibition of gelatinases (MMP-2 and MMP-9) by Withania somnifera phytochemicals confers neuroprotection in stroke: an in silico analysis. Interdiscip Sci Comput Life Sci 10:722–733.  https://doi.org/10.1007/s12539-017-0231-x CrossRefGoogle Scholar
  29. Kumar G, Paliwal P, Patnaik N, Patnaik R (2017) Withania somnifera phytochemicals confer neuroprotection by selective inhibition of nNos: an in silico study to search potent and selective inhibitors for human nNOS. J Theor Comput Chem 16:1750042.  https://doi.org/10.1142/S0219633617500420 CrossRefGoogle Scholar
  30. Kumar G, Paliwal P, Mukherjee S, Patnaik N, Krishnamurthy S, Patnaik R (2018) Pharmacokinetics and brain penetration study of chlorogenic acid in rats. Xenobiotica. 49:339–345.  https://doi.org/10.1080/00498254.2018.1445882 CrossRefGoogle Scholar
  31. Lapchak PA (2007) The phenylpropanoid micronutrient chlorogenic acid improves clinical rating scores in rabbits following multiple infarct ischemic strokes: synergism with tissue plasminogen activator. Exp Neurol 205:407–413.  https://doi.org/10.1016/j.expneurol.2007.02.017 CrossRefGoogle Scholar
  32. Lapchak PA (2011) Emerging therapies: pleiotropic multi-target drugs to treat stroke victims. Transl Stroke Res 2:129–135CrossRefGoogle Scholar
  33. Lee JM, Grabb MC, Zipfel GJ, Choi DW (2000) Brain tissue responses to ischemia. J Clin Invest 106:723–731.  https://doi.org/10.1172/JCI11003 CrossRefGoogle Scholar
  34. Lee K, Lee J-S, Jang H-J, Kim SM, Chang MS, Park SH, Kim KS, Bae J, Park JW, Lee B, Choi HY, Jeong CH, Bu Y (2012) Chlorogenic acid ameliorates brain damage and edema by inhibiting matrix metalloproteinase-2 and 9 in a rat model of focal cerebral ischemia. Eur J Pharmacol 689:89–95.  https://doi.org/10.1016/j.ejphar.2012.05.028 CrossRefGoogle Scholar
  35. Li MH, Inoue K, Si HF, Xiong ZG (2011) Calcium-permeable ion channels involved in glutamate receptor-independent ischemic brain injury. In: Acta Pharmacologica SinicaGoogle Scholar
  36. Liu T, Clark RK, McDonnell PC et al (1994) Tumor necrosis factor-alpha expression in ischemic neurons. Stroke. 25:1481–1488.  https://doi.org/10.1161/01.STR.25.7.1481 CrossRefGoogle Scholar
  37. Mărgăritescu O, Mogoantă L, Pirici I et al (2009) Histopathological changes in acute ischemic stroke. Romanian J Morphol EmbryolGoogle Scholar
  38. Martin Y, Avendaño C, Piedras MJ, Krzyzanowska A (2010) Evaluation of Evans blue extravasation as a measure of peripheral inflammationGoogle Scholar
  39. Mathers CD, Loncar D (2006) Projections of global mortality and burden of disease from 2002 to 2030. PLoS Med 3.  https://doi.org/10.1371/journal.pmed.0030442
  40. Mattila OS, Strbian D, Saksi J, Pikkarainen TO, Rantanen V, Tatlisumak T, Lindsberg PJ (2011) Cerebral mast cells mediate blood-brain barrier disruption in acute experimental ischemic stroke through perivascular gelatinase activation. Stroke. 42:3600–3605.  https://doi.org/10.1161/STROKEAHA.111.632224 CrossRefGoogle Scholar
  41. Meng X-Y, Zhang H-X, Mezei M, Cui M (2011) Molecular docking: a powerful approach for structure-based drug discovery. Curr Comput Aided Drug Des 7:146–157.  https://doi.org/10.1016/j.biotechadv.2011.08.021.Secreted CrossRefGoogle Scholar
  42. Miao M, Cao L, Li R, Fang X, Miao Y (2017) Protective effect of chlorogenic acid on the focal cerebral ischemia reperfusion rat models. Saudi Pharm J 25:556–563.  https://doi.org/10.1016/j.jsps.2017.04.023 CrossRefGoogle Scholar
  43. Mikami Y, Yamazawa T (2015) Chlorogenic acid, a polyphenol in coffee, protects neurons against glutamate neurotoxicity. Life Sci 139:69–74.  https://doi.org/10.1016/j.lfs.2015.08.005 CrossRefGoogle Scholar
  44. Mishra V, Verma R, Singh N, Raghubir R (2011) The neuroprotective effects of NMDAR antagonist, ifenprodil and ASIC1a inhibitor, flurbiprofen on post-ischemic cerebral injury. Brain Res 1389:152–160.  https://doi.org/10.1016/j.brainres.2011.03.011 CrossRefGoogle Scholar
  45. Mohammadi MT (2016) Overproduction of nitric oxide intensifies brain infarction and cerebrovascular damage through reduction of claudin-5 and ZO-1 expression in striatum of ischemic brain. Pathol Res Pract 212:959–964.  https://doi.org/10.1016/j.prp.2015.12.009 CrossRefGoogle Scholar
  46. Mohammadi MT, Shid-Moosavi SM, Dehghani GA (2012) Contribution of nitric oxide synthase (NOS) in blood-brain barrier disruption during acute focal cerebral ischemia in normal rat. Pathophysiology. 19:13–20.  https://doi.org/10.1016/j.pathophys.2011.07.003 CrossRefGoogle Scholar
  47. Moore PK, Wallace P, Gaffen Z, Hart SL, Babbedge RC (1993) Characterization of the novel nitric oxide synthase inhibitor 7-nitro indazole and related indazoles: antinociceptive and cardiovascular effects. Br J Pharmacol 110:219–224.  https://doi.org/10.1111/j.1476-5381.1993.tb13795.x CrossRefGoogle Scholar
  48. Nabavi SF, Tejada S, Setzer WN, Gortzi O, Sureda A, Braidy N, Daglia M, Manayi A, Nabavi SM (2017) Chlorogenic acid and mental diseases: from chemistry to medicine. Curr Neuropharmacol 15:471–479.  https://doi.org/10.2174/1570159X14666160325120625 CrossRefGoogle Scholar
  49. Nagahori T, Nishijima M, Endo S, Takaku A, Iwasaki Y (1994) Ischemic brain damage induced by repeated brief occlusions of bilateral common carotid artery in rats. Tohoku J Exp Med 172:253–262CrossRefGoogle Scholar
  50. Nanri K, Montécot C, Springhetti V et al (1998) The selective inhibitor of neuronal nitric oxide synthase, 7-nitroindazole, reduces the delayed neuronal damage due to forebrain ischemia in rats • editorial comment. Stroke 29:1248–1254.  https://doi.org/10.1161/01.STR.29.6.1248 CrossRefGoogle Scholar
  51. Nirogi R, Kandikere V, Mudigonda K, Bhyrapuneni G, Muddana N, Saralaya R, Benade V (2009) A simple and rapid method to collect the cerebrospinal fluid of rats and its application for the assessment of drug penetration into the central nervous system. J Neurosci Methods 178:116–119.  https://doi.org/10.1016/j.jneumeth.2008.12.001 CrossRefGoogle Scholar
  52. O’Mahony D, Kendall MJ (1999) Nitric oxide in acute ischaemic stroke: a target for neuroprotection. J Neurol Neurosurg Psychiatry 67:1–3CrossRefGoogle Scholar
  53. Ogata J, Fujishima M, Morotomi Y, Omae T (1976) Cerebral infarction following bilateral carotid artery ligation in normotensive and spontaneously hypertensive rats: a pathological study. Stroke. 7:54–60.  https://doi.org/10.1161/01.STR.7.1.54 CrossRefGoogle Scholar
  54. Parmentier S, G a B, Lerouet D et al (1999) Selective inhibition of inducible nitric oxide synthase prevents ischaemic brain injury. Br J Pharmacol 127:546–552.  https://doi.org/10.1038/sj.bjp.0702549 CrossRefGoogle Scholar
  55. Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM, Meng EC, Ferrin TE (2004) UCSF chimera—a visualization system for exploratory research and analysis. J Comput Chem 25:1605–1612.  https://doi.org/10.1002/jcc.20084 CrossRefGoogle Scholar
  56. Rebai O, Belkhir M, Sanchez-Gomez MV, Matute C, Fattouch S, Amri M (2017) Differential molecular targets for neuroprotective effect of chlorogenic acid and its related compounds against glutamate induced excitotoxicity and oxidative stress in rat cortical neurons. Neurochem Res 42:3559–3572.  https://doi.org/10.1007/s11064-017-2403-9 CrossRefGoogle Scholar
  57. Rudnitskaya A, Török B, Török M (2010) Molecular docking of enzyme inhibitors: a computational tool for structure-based drug design. Biochem Mol Biol Educ 38:261–265.  https://doi.org/10.1002/bmb.20392 CrossRefGoogle Scholar
  58. Sadoshima S, Fujishima M, Ogata J, Ibayashi S, Shiokawa O, Omae T (1983) Disruption of blood-brain barrier following bilateral carotid artery occlusion in spontaneously hypertensive rats. A quantitative study. Stroke. 14:876–882.  https://doi.org/10.1161/01.STR.14.6.876 CrossRefGoogle Scholar
  59. Sanner MF (1999) Python: a programming language for software integration and development. J Mol Graph Model 17:55–84.  https://doi.org/10.1016/S1093-3263(99)99999-0 CrossRefGoogle Scholar
  60. Sato Y, Itagaki S, Kurokawa T, Ogura J, Kobayashi M, Hirano T, Sugawara M, Iseki K (2011) In vitro and in vivo antioxidant properties of chlorogenic acid and caffeic acid. Int J Pharm 403:136–138.  https://doi.org/10.1016/j.ijpharm.2010.09.035 CrossRefGoogle Scholar
  61. Schilichting CLR, Lima KCM, Cestari LA et al (2004) Validation of a simple and inexpensive method for the quantitation of infarct in the rat brain. Braz J Med Biol Res 37:511–521.  https://doi.org/10.1590/S0100-879X2004000400008 CrossRefGoogle Scholar
  62. Shen W, Qi R, Zhang J, Wang Z, Wang H, Hu C, Zhao Y, Bie M, Wang Y, Fu Y, Chen M, Lu D (2012) Chlorogenic acid inhibits LPS-induced microglial activation and improves survival of dopaminergic neurons. Brain Res Bull 88:487–494.  https://doi.org/10.1016/j.brainresbull.2012.04.010 CrossRefGoogle Scholar
  63. Simard JM, Kent TA, Chen M, Tarasov KV, Gerzanich V (2007) Brain oedema in focal ischaemia: molecular pathophysiology and theoretical implications. Lancet Neurol 6:258–268CrossRefGoogle Scholar
  64. Singh SS, Rai SN, Birla H, Zahra W, Kumar G, Gedda MR, Tiwari N, Patnaik R, Singh RK, Singh SP (2018) Effect of chlorogenic acid supplementation in MPTP-intoxicated mouse. Front Pharmacol 9:757CrossRefGoogle Scholar
  65. Sun Y, Xu Y, Geng L (2015) Caspase-3 inhibitor prevents the apoptosis of brain tissue in rats with acute cerebral infarction. Exp Ther Med 10:133–138.  https://doi.org/10.3892/etm.2015.2462 CrossRefGoogle Scholar
  66. Suzuki M, Tabuchi M, Ikeda M, Tomita T (2002) Concurrent formation of peroxynitrite with the expression of inducible nitric oxide synthase in the brain during middle cerebral artery occlusion and reperfusion in rats. Brain Res 951:113–120.  https://doi.org/10.1016/S0006-8993(02)03145-1 CrossRefGoogle Scholar
  67. Türeyen K, Vemuganti R, Sailor KA, Dempsey RJ (2004) Infarct volume quantification in mouse focal cerebral ischemia: a comparison of triphenyltetrazolium chloride and cresyl violet staining techniques. J Neurosci Methods 139:203–207.  https://doi.org/10.1016/j.jneumeth.2004.04.029 CrossRefGoogle Scholar
  68. Tuttolomondo A, Pecoraro R, Pinto A (2014) Studies of selective TNF inhibitors in the treatment of brain injury from stroke and trauma: a review of the evidence to date. Drug Des Devel Ther.  https://doi.org/10.2147/DDDT.S67655
  69. Van Den Tweel ERW, Peeters-Scholte CMPCD, Van Bel F et al (2002) Inhibition of nNOS and iNOS following hypoxia-ischaemia improves long-term outcome but does not influence the inflammatory response in the neonatal rat brain. In: Developmental NeuroscienceGoogle Scholar
  70. Varma AK, Patil R, Das S et al (2010) Optimized hydrophobic interactions and hydrogen bonding at the target-ligand interface leads the pathways of drug-designing. PLoS One 5:e12029.  https://doi.org/10.1371/journal.pone.0012029 CrossRefGoogle Scholar
  71. Wang X, Lin F, Gao Y, Lei H (2015) Bilateral common carotid artery occlusion induced brain lesions in rats: a longitudinal diffusion tensor imaging study. Magn Reson Imaging 33:551–558.  https://doi.org/10.1016/j.mri.2015.02.010 CrossRefGoogle Scholar
  72. Wang L, Liu H, Zhang L, Wang G, Zhang M, Yu Y (2017) Neuroprotection of dexmedetomidine against cerebral ischemia-reperfusion injury in rats: involved in inhibition of NF-κB and inflammation response. Biomol Ther 25:383–389.  https://doi.org/10.4062/biomolther.2015.180 CrossRefGoogle Scholar
  73. Wu QJ, Tymianski M (2018) Targeting nmda receptors in stroke: new hope in neuroprotection Tim bliss. Mol BrainGoogle Scholar
  74. Wu X, Wang R, Jiang Q, Wang S, Yao Y, Shao L (2014) Determination of amino acid neurotransmitters in rat hippocampi by HPLC-UV using NBD-F as a derivative. Biomed Chromatogr 28:459–462.  https://doi.org/10.1002/bmc.3062 CrossRefGoogle Scholar
  75. Xing C, Arai K, Lo EH, Hommel M (2012) Pathophysiologic cascades in ischemic stroke. Int J Stroke 7:378–385CrossRefGoogle Scholar
  76. Xu L, Li Y, Fu Q, Ma S (2014) Perillaldehyde attenuates cerebral ischemia-reperfusion injury-triggered overexpression of inflammatory cytokines via modulating Akt/JNK pathway in the rat brain cortex. Biochem Biophys Res Commun 454:65–70.  https://doi.org/10.1016/j.bbrc.2014.10.025 CrossRefGoogle Scholar
  77. Zheng L, Ding J, Wang J, Zhou C, Zhang W (2016) Effects and mechanism of action of inducible nitric oxide synthase on apoptosis in a rat model of cerebral ischemia-reperfusion injury. Anat Rec 299:246–255.  https://doi.org/10.1002/ar.23295 CrossRefGoogle Scholar
  78. Zhu Y, Yang G-Y, Ahlemeyer B, Pang L, Che XM, Culmsee C, Klumpp S, Krieglstein J (2002) Transforming growth factor-β1 increases bad phosphorylation and protects neurons against damage. J Neurosci 22:3898–3909.  https://doi.org/10.1523/JNEUROSCI.22-10-03898.2002 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Electrophysiology Lab, School of Biomedical EngineeringIndian Institute of Technology (Banaras Hindu University)VaranasiIndia
  2. 2.Department of Pharmaceutical Engineering and TechnologyIndian Institute of Technology (Banaras Hindu University)VaranasiIndia
  3. 3.Department of Biochemistry, Institute of ScienceBanaras Hindu UniversityVaranasiIndia

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