Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 391, Issue 9, pp 987–1001 | Cite as

Ellagic acid prevents dementia through modulation of PI3-kinase-endothelial nitric oxide synthase signalling in streptozotocin-treated rats

  • Manish Kumar
  • Nitin Bansal
Original Article


Ellagic acid (EGA)-enriched dietary supplements are widely acclaimed, owing to its versatile bioactivities. Previously, we reported that chronic administration of EGA prevented the impairment of cognitive abilities in rats using the intracerebroventricular-administered streptozotocin (STZ-ICV) model of Alzheimer’s disease. Impairment of phosphoinositide 3 (PI3)-kinase-regulated endothelial nitric oxide synthase (eNOS) activity by central administration of STZ in rodents instigates dementia. The aim of the present study was to delineate the role of PI3-kinase-eNOS activity in the prevention of STZ-ICV-induced memory dysfunctions by EGA. The Morris water maze and elevated plus maze tests were conducted, and brain oxidative stress markers (TBARS, GSH, SOD, CAT), nitrite, acetylcholinesterase (AChE), LDH, TNF-α and eNOS were quantified. Administration of EGA (35 mg/k, p.o.) for 4 weeks daily attenuated the STZ-ICV (3 mg/kg)-triggered increase of brain oxidative stress, nitrite and TNF-α levels; AChE and LDH activity; and decline of brain eNOS activity. The memory restoration by EGA in STZ-ICV-treated rats was conspicuously impaired by N(G)-nitro-l-arginine methyl ester (L-NAME) (20 mg/kg, 28 days) and wortmannin (5 μg/rat; ICV) treatments. Wortmannin (PI3-kinase inhibitor) and L-NAME groups manifested elevated brain oxidative stress, TNF-α content and AChE and LDH activity and diminished nitrite content. L-NAME (arginine-based competitive eNOS inhibitor) enhanced the eNOS expression (not activity) whereas wortmannin reduced the brain eNOS levels in EGA- and STZ-ICV-treated rats. However, the L-NAME group exhibited superior cognitive abilities in comparison to the wortmannin group. It can be concluded that EGA averted the memory deficits by precluding the STZ-ICV-induced loss of PI3-kinase-eNOS signalling in the brain of rats.


Ellagic acid PI3-kinase-endothelial nitric oxide synthase (eNOS) Memory Wortmannin Streptozotocin Acetylcholinesterase 



The authors are thankful to the management of ASBASJSM College of Pharmacy, Bela, for providing the necessary research facilities and IKG Punjab Technical University, Kapurthala.

Authors’ contribution

Prof. (Dr.) Nitin Bansal designed this study. Manish Kumar (PhD research scholar) conducted the research and analysed and interpreted the data. Both authors wrote the initial and final draft of the article.

Funding information

This research was assisted by a grant from AICTE, New Delhi (India), under Research Promotion Scheme.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Agrawal R, Tyagi E, Shukla R, Nath C (2011) Insulin receptor signaling in rat hippocampus: a study in STZ (ICV) induced memory deficit model. Eur Neuropsychopharmacol 21:261–273. CrossRefPubMedGoogle Scholar
  2. Austin SA, Santhanam AV, Hinton DJ, Choi DS, Katusic ZS (2013) Endothelial nitric oxide deficiency promotes Alzheimer’s disease pathology. J Neurochem 127:691–700. CrossRefPubMedPubMedCentralGoogle Scholar
  3. Baki L, Shioi J, Wen P, Shao Z, Schwarzman A, Gama-Sosa M, Neve R, Robakis N (2004) PS1 activates PI3K thus inhibiting GSK-3 activity and tau overphosphorylation: effects of FAD mutations. EMBO J 23:2586–2596. CrossRefPubMedPubMedCentralGoogle Scholar
  4. Bansal N, Yadav P, Kumar M (2017) Ellagic acid administration negated the development of streptozotocin-induced memory deficit in rats. Drug Res (Stuttg) 67:425–431. CrossRefGoogle Scholar
  5. Bedse G, Di Domenico F, Serviddio G, Cassano T (2015) Aberrant insulin signaling in Alzheimer’s disease: current knowledge. Front Neurosci 9:204. CrossRefPubMedPubMedCentralGoogle Scholar
  6. Calsolaro V, Edison P (2016) Neuroinflammation in Alzheimer’s disease: current evidence and future directions. Alzheimers Dement 12:719–732. CrossRefPubMedGoogle Scholar
  7. Chen HT, Ruan NY, Chen JC, Lin TY (2012) Dopamine D2 receptor-mediated Akt/PKB signalling: initiation by the D2S receptor and role in quinpirole-induced behavioural activation. ASN Neuro 4:371–382. CrossRefPubMedGoogle Scholar
  8. Claiborne A (1985) Catalase activity. In: Greenwald RA (ed) CRC handbook of methods for oxygen radical research, 3rd edn. CRC, Boca Raton, pp 283–284Google Scholar
  9. Ding Y, Zhang B, Zhou K, Chen M, Wang M, Jia Y, Song Y, Li Y, Wen A (2014) Dietary ellagic acid improves oxidant-induced endothelial dysfunction and atherosclerosis: role of Nrf2 activation. Int J Cardiol 175:508–514. CrossRefPubMedGoogle Scholar
  10. Doens D, Fernandez PL (2014) Microglia receptors and their implications in the response to amyloid β for Alzheimer’s disease pathogenesis. J Neuroinflammation 11:48. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Ellman GL (1959) Tissue sulfhydryl groups. Arch Biochem Biophys 82:70–77CrossRefPubMedGoogle Scholar
  12. Ellman GL, Courtney KD, VJr A, Feather-Stone RM (1961) A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–95CrossRefPubMedGoogle Scholar
  13. Ferreira-Vieira TH, Guimaraes IM, Silva FR, Ribeiro FM (2016) Alzheimer’s disease: targeting the cholinergic system. Curr Neuropharmacol 14:101–115. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Franke TF (2008) PI3K/Akt: getting it right matters. Oncogene 27:6473–6488. CrossRefPubMedGoogle Scholar
  15. Gao C, Liu Y, Jiang Y, Ding J, Li L (2014) Geniposide ameliorates learning memory deficits, reduces tau phosphorylation and decreases apoptosis via GSK3β pathway in streptozotocin-induced Alzheimer rat model. Brain Pathol 24:261–269. CrossRefPubMedGoogle Scholar
  16. Guha M, Mackman N (2002) The phosphatidylinositol 3-kinase-Akt pathway limits lipopolysaccharide activation of signaling pathways and expression of inflammatory mediators in human monocytic cells. J Biol Chem 277:32124–32132. CrossRefPubMedGoogle Scholar
  17. Harry GJ, Sills R, Schlosser MJ, Maier WE (2001) Neurodegeneration and glia response in rat hippocampus following nitro-L-arginine methyl ester (L-NAME). Neurotox Res 3:307–319CrossRefPubMedGoogle Scholar
  18. Horecker BL, Kornberg A (1948) The extinction coefficient of the reduced band of pyridine nucleotides. J Biol Chem 175:385–390PubMedGoogle Scholar
  19. Kiasalari Z, Heydarifard R, Khalili M, Afshin-Majd S, Baluchnejadmojarad T, Zahedi E, Sanaierad A, Roghani M (2017) Ellagic acid ameliorates learning and memory deficits in a rat model of Alzheimer’s disease: an exploration of underlying mechanisms. Psychopharmacology 234:1841–1852. CrossRefPubMedGoogle Scholar
  20. Kong J, Ren G, Jia N, Wang Y, Zhang H, Zhang W, Chen B, Cao Y (2013) Effects of nicorandil in neuroprotective activation of PI3K/AKT pathways in a cellular model of Alzheimer’s disease. Eur Neurol 70:233–241. CrossRefPubMedGoogle Scholar
  21. Landete JM (2011) Ellagitannins, ellagic acid and their derived metabolites: a review about source, metabolism, functions and health. Food Res Int 44:1150–1160. CrossRefGoogle Scholar
  22. Lee HJ, Ryu JM, Jung YH, Lee SJ, Kim JY, Lee SH, Hwang IK, Seong JK, Han HJ (2016) High glucose upregulates BACE1-mediated Aβ production through ROS-dependent HIF-1α and LXRα/ABCA1-regulated lipid raft reorganization in SK-N-MC cells. Sci Rep 6:36746. CrossRefPubMedPubMedCentralGoogle Scholar
  23. Li ST, Pan J, Hua XM, Liu H, Shen S, Liu JF, Li B, Tao BB, Ge XL, Wang XH, Shi JH, Wang XQ (2014) Endothelial nitric oxide synthase protects neurons against ischemic injury through regulation of brain-derived neurotrophic factor expression. CNS Neurosci Ther 20:154–164. CrossRefPubMedGoogle Scholar
  24. Lipinska L, Klewicka E, Sojka M (2014) The structure, occurrence and biological activity of ellagitannins: a general review. Acta Sci Pol Technol Aliment 13:289–299CrossRefPubMedGoogle Scholar
  25. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  26. Mansouri MT, Farbood Y, Naghizadeh B, Shabani S, Mirshekar MA, Sarkaki A (2016) Beneficial effects of ellagic acid against animal models of scopolamine- and diazepam-induced cognitive impairments. Pharm Biol 54:1947–1953. CrossRefPubMedGoogle Scholar
  27. Meng Y, Wang W, Kang J, Wang X, Sun L (2017) Role of the PI3K/AKT signalling pathway in apoptotic cell death in the cerebral cortex of streptozotocin-induced diabetic rats. Exp Ther Med 13:2417–2422. CrossRefPubMedPubMedCentralGoogle Scholar
  28. Mohan S, Wu CC, Shin S, Fung HL (2012) Continuous exposure to L-arginine induces oxidative stress and physiological tolerance in cultured human endothelial cells. Amino Acids 43:1179–1188. CrossRefPubMedGoogle Scholar
  29. Moosavi M, Abbasi L, Zarifkar A, Rastegar K (2014) The role of nitric oxide in spatial memory stages, hippocampal ERK and CaMKII phosphorylation. Pharmacol Biochem Behav 122:164–172. CrossRefPubMedGoogle Scholar
  30. Morris RGM (1984) Development of a water-maze procedure for studying spatial learning in the rats. J Neurosci Methods 11:47–60CrossRefPubMedGoogle Scholar
  31. Murphy MP (1999) Nitric oxide and cell death. Biochim Biophys Acta 1411:401–414CrossRefPubMedGoogle Scholar
  32. Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358CrossRefPubMedGoogle Scholar
  33. Ou HC, Lee WJ, Lee SD, Huang CY, Chiu TH, Tsai KL, Hsu WC, Sheu WH (2010) Ellagic acid protects endothelial cells from oxidized low-density lipoprotein-induced apoptosis by modulating the PI3K/Akt/eNOS pathway. Toxicol Appl Pharmacol 248:134–143. CrossRefPubMedGoogle Scholar
  34. Paul V, Ekambaram P (2011) Involvement of nitric oxide in learning & memory processes. Indian J Med Res 133:471–478PubMedPubMedCentralGoogle Scholar
  35. Paxinos G, Watson CR, Emson PC (1980) AChE-stained horizontal sections of the rat brain in stereotaxic coordinates. J Neurosci Methods 3:129–149CrossRefPubMedGoogle Scholar
  36. Petanceska SS, Gandy S (1999) The phosphatidylinositol 3-kinase inhibitor wortmannin alters the metabolism of the Alzheimer’s amyloid precursor protein. J Neurochem 73:2316–2320CrossRefPubMedGoogle Scholar
  37. Prast H, Fischer H, Werner E, Werner-Felmayer G, Philippu A (1995) Nitric oxide modulates the release of acetylcholine in the ventral striatum of the freely moving rat. Naunyn Schmiedeberg’s Arch Pharmacol 352:67–73CrossRefGoogle Scholar
  38. Provias J, Jeynes B (2008) Neurofibrillary tangles and senile plaques in Alzheimer’s brains are associated with reduced capillary expression of vascular endothelial growth factor and endothelial nitric oxide synthase. Curr Neurovasc Res 5:199–205. CrossRefPubMedGoogle Scholar
  39. Rajasekar N, Nath C, Hanif K, Shukla R (2017) Intranasal insulin improves cerebral blood flow, Nrf-2 expression and BDNF in STZ (ICV)-induced memory impaired rats. Life Sci 173:1–10. CrossRefPubMedGoogle Scholar
  40. Resende RR, Adhikari A (2009) Cholinergic receptor pathways involved in apoptosis, cell proliferation and neuronal differentiation. Cell Commun Signal 7:20. CrossRefPubMedPubMedCentralGoogle Scholar
  41. Rickard NS, Gibbs ME, Ng KT (1999) Inhibition of the endothelial isoform of nitric oxide synthase impairs long-term memory formation in the chick. Learn Mem 6:458–466CrossRefPubMedPubMedCentralGoogle Scholar
  42. Sastry KV, Moudgal RP, Mohan J, Tyagi JS, Rao GS (2002) Spectrophotometric determination of serum nitrite and nitrate by copper-cadmium alloy. Anal Biochem 306:79–82CrossRefPubMedGoogle Scholar
  43. Sharma N, Deshmukh R, Bedi KL (2010) SP600125, a competitive inhibitor of JNK attenuates streptozotocin induced neurocognitive deficit and oxidative stress in rats. Pharmacol Biochem Behav 96:386–394. CrossRefPubMedGoogle Scholar
  44. Son H, Hawkins RD, Martin K, Kiebler M, Huang PL, Fishman MC, Kandel ER (1996) Long-term potentiation is reduced in mice that are doubly mutant in endothelial and neuronal nitric oxide synthase. Cell 87:1015–1023CrossRefPubMedGoogle Scholar
  45. Traystman RJ, Moore LE, Helfaer MA, Davis S, Banasiak K, Williams M, Hurn PD (1995) Nitro-L-arginine analogues. Dose- and time-related nitric oxide synthase inhibition in brain. Stroke 26:864–869CrossRefPubMedGoogle Scholar
  46. Tyagi E, Agrawal R, Nath C, Shukla R (2010) Cholinergic protection via alpha7 nicotinic acetylcholine receptors and PI3K-Akt pathway in LPS-induced neuroinflammation. Neurochem Int 56:135–142. CrossRefPubMedGoogle Scholar
  47. Utkan T, Yazir Y, Karson A, Bayramgurler D (2015) Etanercept improves cognitive performance and increases eNOS and BDNF expression during experimental vascular dementia in streptozotocin-induced diabetes. Curr Neurovasc Res 12:135–146. CrossRefPubMedGoogle Scholar
  48. Vivanco I, Sawyers CL (2002) The phosphatidylinositol 3-kinase AKT pathway in human cancer. Nat Rev Cancer 2:489–501. CrossRefPubMedGoogle Scholar
  49. Vorhees CV, Williams MT (2006) Morris water maze: procedures for assessing spatial and related forms of learning and memory. Nat Protoc 1:848–858CrossRefPubMedPubMedCentralGoogle Scholar
  50. Winterbourn CC, Hawkins RE, Brian M, Carrell RW (1975) The estimation of red cell superoxide dismutase activity. J Lab Clin Med 85:337–341PubMedGoogle Scholar
  51. Yang Y, Ma D, Xu W, Chen F, Du T, Yue W, Shao S, Yuan G (2016) Exendin-4 reduces tau hyperphosphorylation in type 2 diabetic rats via increasing brain insulin level. Mol Cell Neurosci 70:68–75. CrossRefPubMedGoogle Scholar
  52. Yoshida T, Amakura Y, Yoshimura M (2010) Structural features and biological properties of ellagitannins in some plant families of the order Myrtales. Int J Mol Sci 11:79–106. CrossRefPubMedPubMedCentralGoogle Scholar
  53. Zhao Y, Hu X, Liu Y, Dong S, Wen Z, He W, Zhang S, Huang Q, Shi M (2017) ROS signaling under metabolic stress: cross-talk between AMPK and AKT pathway. Mol Cancer 16:79. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.IKG Punjab Technical UniversityKapurthalaIndia
  2. 2.Department of PharmacologyASBASJSM College of PharmacyRoparIndia

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