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PHARMASPIRE - Volume 10, Issue 3, July - September, 2018

Pages: 132-140

Date of Publication: 14-Jun-2022

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Possible mechanism of sustained 5-adenosine monophosphate-activated protein kinase activator associated with neuronal nitric oxide synthase inhibition in global cerebral ischemia-induced neurodegeneration in rats

Author: Himanshi Khera, Sidharth Mehan, Rajesh Dudi

Category: Pharmaceutics


Global cerebral ischemia (GCI) is a clinical condition that causes a deprivation of blood supply and energy in the brain due to blockade of carotid arteries. The decreased level of oxygen and glucose causes various cellular changes leading to excitotoxicity and oxidative damage. Cerebral ischemia leads to cell death in CA1 region of the hippocampus, which occurs 3–4 days after an initial ischemic insult and neurodegeneration is evident by 3 days of reperfusion, and neuronal death culminates by 6 days. GCI was induced by bilateral carotid artery occlusion for 10 min. 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) (50 µg and 100 µg) and 3-Br-7-nitroindazole (10 µg, 20 µg, and 50 µg) were administered once a daily for 4 days as a treatment. Cerebral ischemia leads to 5-adenosine monophosphate-activated protein kinase (AMPK) activation, and further administration of AICAR leads to sustained activation of AMPK. Literature has suggested that AMPK activation causes stimulation of mitochondrial biogenesis resulting in an increase in free radical accumulation. In the present study, cerebral ischemia has been noted to cause: Increased malondialdehyde levels, decreased glutathione levels, increased lactate dehydrogenase levels, and decreased acetylcholinesterase levels, and increased nitrate levels in brain homogenate. AMPK activation through AICAR administration was observed to damage cortex and hippocampal regions of brain as are evident from photomicrographs. Thus, it may be concluded from the present study that AMPK activation in ischemic animals leads to neurodegeneration.

Keywords: Global cerebral ischemia, 5-adenosine monophosphate-activated protein kinase, 5-aminoimidazole-4-carboxamide ribonucleotide, excitotoxicity, hippocampus, neurodegeneration


1. Sarkar PK. Degeneration and death of neurons in adult neurodegenerative diseases. Curr Sci 2005;10:764-73.

2. Sharma SS, Gupta S. Neuroprotective effect of mnTMPyP, a superoxide dismutase/catalase mimetic in global cerebral ischemia is mediated through reduction of oxidative stress and DNA fragmentation. Eur J Pharmacol 2007;561:72-9.

3. Smith WS. Pathophysiology of focal cerebral ischemia: A therapeutic perspective. JVasc Interv Radiol 2004;15:S3-12.

4. Graham SH, Chen J. Programmed cell death in cerebral ischemia. J Cereb Blood Flow Metab 2001;21:99-109.

5. Ginsberg MD, Busto R. Rodent models of cerebral ischemia. Stroke 1989;20:1627-42.

6. Derouesné C, Cambon H, Yelnik A, Duyckaerts C, Hauw JJ. Infarcts in the middle cerebral artery territory. Pathological study of the mechanisms of death. Acta Neurol Scand 1993;87:361-6.

7. Lipton P. Ischemic cell death in brain neurons. Physiol Rev 1999;79:1431-568.

8. Muralikrishna Adibhatla R, Hatcher JF. Phospholipase A2, reactive oxygen species, and lipid peroxidation in cerebral ischemia. Free Radic Biol Med 2006;40:376-87.

9. Gupta YK, Briyal S. Animal models of cerebral ischemia for evaluation of drugs. Indian J Physiol Pharmacol 2004;48:379-94.

10. Kirino T, Sano K. Fine strucStural nature of delayed neuronal death following ischemia in the gerbil hippocampus. Acta Neuropathol 1984;62:209-18.

11. Corbett D, Nurse S. The problem of assessing effective neuroprotection in experimental cerebral ischemia. Prog Neurobiol 1998;54:531-48.

12. Wang P, Wang WP, Sun-Zhang, Wang HX, Yan-Lou, Fan YH. Impaired spatial learning related with decreased expression of calcium/calmodulin dependent protein kinase-II alpha and cAMP-response element binding protein in the pentylenetetrazol-kindled rats. Brain Res 2008;1238:108-17.

13. Ramamurthy S, Ronnett GV. Developing a head for energy sensing: AMPactivated protein kinase as a multifunctional metabolic sensor in the brain. J Physiol 2006;574:85-93.

14. Hardie DG. AMP-activated protein kinase as a drug target. Annu Rev Pharmacol Toxicol 2007;47:185-210.

15. Almeida A, Moncada S, Bolaños JP. Nitric oxide switches on glycolysis through the AMP protein kinase and 6-phosphofructo-2-kinase pathway. Nat Cell Biol 2004;6:45-51.

16. Hardie DG, Frenguelli BG. A neural protection racket: AMPK and the GABA (B) receptor. Neuron 2007;53:159-62.

17. Hawley SA, Davison M, Woods A, Davies SP, Beri RK, Carling D, et al. Characterization of the AMP-activated protein kinase kinase from rat liver and identification of threonine 172 as the major site at which it phosphorylates AMP-activated protein kinase. J Biol Chem 1996;271:27879-87.

18. Bloomgarden ZT. Fat metabolism and diabetes: 2003 American diabetes association postgraduate course. Diabetes Care 2003;26:2198-203.

19. Henin N, Vincent MF, Gruber HE, Van den Berghe G. Inhibition of fatty acid and cholesterol synthesis by stimulation of AMP-activated protein kinase. FASEB J 1995;9:541-6.

20. Kurth-Kraczek EJ, Hirshman MF, Goodyear LJ, Winder WW. 5’ AMP-activated protein kinase activation causes GLUT4 translocation in skeletal muscle. Diabetes 1999;48:1667-71.

21. Culmsee C, Monnig J, Kemp BE, Mattson MP. AMP-activated protein kinase is highly expressed in neurons in the developing rat brain and promotes neuronal survival following glucose deprivation. J Mol Neurosci 2001;17:45-58.

22. Tzatsos A, Tsichlis PN. Energy depletion inhibits phosphatidylinositol 3-kinase/ Akt signaling and induces apoptosis via AMP-activated protein kinase-dependent phosphorylation of IRS-1 at ser-794. J Biol Chem 2007;282:18069-82.

23. Petito CK, Feldmann E, Pulsinelli WA, Plum F. Delayed hippocampal damage in humans following cardiorespiratory arrest. Neurology 1987;37:1281-6.

24. Smith ML, Auer RN, Siesjö BK. The density and distribution of ischemic brain injury in the rat following 2-10 min of forebrain ischemia. Acta Neuropathol 1984;64:319-32.

25. Smith ML, Bendek G, Dahlgren N, Rosén I, Wieloch T, Siesjö BK, et al. Models for studying long-term recovery following forebrain ischemia in the rat 2. A 2-vessel occlusion model. Acta Neurol Scand 1984;69:385-401.

26. Kumar A, Naidu PS, Seghal N, Padi SS. Neuroprotective effects of resveratrol against intracerebroventricular colchicine-induced cognitive impairment and oxidative stress in rats. Pharmacology 2007;79:17-26.

27. von Euler M, Bendel O, Bueters T, Sandin J, von Euler G. Profound but transient deficits in learning and memory after global ischemia using a novel water maze test. Behav Brain Res 2006;166:204-10.

28. Ramos-Zúñiga R, Gómez PU, Navarro Ruiz A, Luquín de AS, García-Estrada J. Locomotor activity is a predictive test after global ischemia-reperfusion in mongolian gerbils. Minim Invasive Neurosurg 2008;51:87-90.

29. Fu X, Wan S, Lyu YL, Liu LF, Qi H. Etoposide induces ATM-dependent mitochondrial biogenesis through AMPK activation. PLoS One 2008;3:e2009.

30. Gao G, Widmer J, Stapleton D, Teh T, Cox T, Kemp BE, et al. Catalytic subunits of the porcine and rat 5’-AMP-activated protein kinase are members of the SNF1 protein kinase family. Biochim Biophys Acta 1995;1266:73-82.

31. Turnley AM, Stapleton D, Mann RJ, Witters LA, Kemp BE, Bartlett PF, et al. Cellular distribution and developmental expression of AMP-activated protein kinase isoforms in mouse central nervous system. J Neurochem 1999;72:1707-16.

32. Kuan CY, Burke RE. Targeting the JNK signaling pathway for stroke and Parkinson’s diseases therapy. Curr Drug Targets CNS Neurol Disord 2005;4:63-7.

33. Okuno S, Saito A, Hayashi T, Chan PH. The c-jun N-terminal protein kinase signaling pathway mediates bax activation and subsequent neuronal apoptosis through interaction with bim after transient focal cerebral ischemia. J Neurosci 2004;24:7879-87