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Activation of AMP-activated protein kinase by metformin protects against global cerebral ischemia in male rats: Interference of AMPK/PGC-1α pathway

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Abstract

Here, we have investigated the effect of metformin pretreatment in the rat models of global cerebral ischemia. Cerebral ischemia which leads to brain dysfunction is one of the main causes of neurodegeneration and death worldwide. Metformin is used in clinical drug therapy protocols of diabetes. It is suggested that metformin protects cells under hypoxia and ischemia in non-neuronal contexts. Protective effects of metformin may be modulated via activating the AMP activated protein kinase (AMPK). Our results showed that induction of 30 min global cerebral I/R injury using 4-vesseles occlusion model led to significant cell death in the rat brain. Metformin pretreatment (200 mg kg/once/day, p.o., 2 weeks) attenuated apoptotic cell death and induced mitochondrial biogenesis proteins in the ischemic rats, analyzed using histological and Western blot assays. Besides, inhibition of AMPK by compound c showed that metformin resulted in apoptosis attenuation via AMPK activation. Interestingly, AMPK activation was also involved in the induction of mitochondrial biogenesis proteins using metformin, inhibition of AMPK by compound c reversed such effect, further supporting the role of AMPK upstream of mitochondrial biogenesis proteins. In summary, Metformin pretreatment is able to modulate mitochondrial biogenesis and apoptotic cell death pathways through AMPK activation in the context of global cerebral ischemia, conducting the outcome towards neuroprotection.

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References

  • Atherton PJ, Babraj J, Smith K, Singh J, Rennie MJ, Wackerhage H (2005) Selective activation of AMPK-PGC-1alpha or PKB-TSC2-mTOR signaling can explain specific adaptive responses to endurance or resistance training-like electrical muscle stimulation. FASEB J 19:786–788

    CAS  PubMed  Google Scholar 

  • Balkaya M, Krober JM, Rex A, Endres M (2012) Assessing post-stroke behavior in mouse models of focal ischemia. J Cereb Blood Flow Metab 33:330–338

    Article  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Capano M, Crompton M (2006) Bax translocates to mitochondria of heart cells during simulated ischaemia: involvement of AMP-activated and p38 mitogen-activated protein kinases. Biochem J 395:57–64

    Article  CAS  PubMed  Google Scholar 

  • Chen SD, Yang DI, Lin TK, Shaw FZ, Liou CW, Chuang YC (2011) Roles of oxidative stress, apoptosis, PGC-1alpha and mitochondrial biogenesis in cerebral ischemia. Int J Mol Sci 12:7199–7215

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Christ-Crain M, Kola B, Lolli F, Fekete C, Seboek D, Wittmann G, Feltrin D, Igreja SC, Ajodha S, Harvey-White J, Kunos G, Muller B, Pralong F, Aubert G, Arnaldi G, Giacchetti G, Boscaro M, Grossman AB, Korbonits M (2008) AMP-activated protein kinase mediates glucocorticoid-induced metabolic changes: a novel mechanism in Cushing’s syndrome. FASEB J 22(6):1672–1683

    Article  CAS  PubMed  Google Scholar 

  • Diler A, Ziylan Y, Uzum G, Lefauconnier J, Seylaz J, Pinard E (2002) Passage of spermidine across the blood–brain barrier in short recirculation periods following global cerebral ischemia: effects of mild hyperthermia. Neurosci Res 43:335–342

    Article  CAS  PubMed  Google Scholar 

  • El-Mir MY, Detaille D, Villanueva GR, Delgado-Esteban M, Guigas B, Attia S, Fontaine E, Almeida A, Leverve X (2008) Neuroprotective role of antidiabetic drug metformin against apoptotic cell death in primary cortical neurons. J Mol Neurosci 34:77–87

    Article  CAS  PubMed  Google Scholar 

  • Gundewar S, Calvert JW, Jha S, Toedt-Pingel I, Ji SY, Nunez D, Ramachandran A, Anaya-Cisneros M, Tian R, Lefer DJ (2009) Activation of AMP-activated protein kinase by metformin improves left ventricular function and survival in heart failure. Circ Res 104:403–411

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Gupta YK, Briyal S, Sharma U, Jagannathan NR, Gulati A (2005) Effect of endothelin antagonist (TAK-044) on cerebral ischemic volume, oxidative stress markers and neurobehavioral parameters in the middle cerebral artery occlusion model of stroke in rats. Life Sci 77:15–27

    Article  CAS  PubMed  Google Scholar 

  • Harada S, Fujita-Hamabe W, Tokuyama S (2010) The importance of regulation of blood glucose levels through activation of peripheral 5′-AMP-activated protein kinase on ischemic neuronal damage. Brain Res 1351:254–263

    Article  CAS  PubMed  Google Scholar 

  • Horman S, Vertommen D, Heath R, Neumann D, Mouton V, Woods A, Schlattner U, Wallimann T, Carling D, Hue L, Rider MH (2006) Insulin antagonizes ischemia-induced Thr172 phosphorylation of AMP-activated protein kinase alpha-subunits in heart via hierarchical phosphorylation of Ser485/491. J Biol Chem 281:5335–5340

    Article  CAS  PubMed  Google Scholar 

  • Irrcher I, Adhihetty PJ, Sheehan T, Joseph AM, Hood DA (2003) PPARgamma coactivator-1alpha expression during thyroid hormone- and contractile activity-induced mitochondrial adaptations. Am J Physiol Cell Physiol 284:C1669–C1677

    Article  CAS  PubMed  Google Scholar 

  • Kirpichnikov D, McFarlane SI, Sowers JR (2002) Metformin: an update. Ann Intern Med 137(1):25–33

    Article  CAS  PubMed  Google Scholar 

  • Kristensen JM, Larsen S, Helge JW, Dela F, Wojtaszewski JF (2013) Two weeks of metformin treatment enhances mitochondrial respiration in skeletal muscle of AMPK kinase dead but not wild type mice. PLoS One 8:e53533

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Lee CK, Choi YJ, Park SY, Kim JY, Won KC, Kim YW (2012) Intracerebroventricular injection of metformin induces anorexia in rats. Diabetes Metab J 36(4):293–299. doi:10.4093/dmj.2012.36.4.293

    Article  PubMed Central  PubMed  Google Scholar 

  • Li XM, Yang JM, Hu DH, Hou FQ, Zhao M, Zhu XH, Wang Y, Li JG, Hu P, Chen L, Qin LN, Gao TM (2007) Contribution of downregulation of L-type calcium currents to delayed neuronal death in rat hippocampus after global cerebral ischemia and reperfusion. J Neurosci 27(19):5249–5259

    Article  CAS  PubMed  Google Scholar 

  • Li J, Benashski SE, Venna VR, McCullough LD (2010) Effects of metformin in experimental stroke. Stroke 41(11):2645–2652

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Li W, Qu Z, Prakash R, Chung C, Ma H, Hoda MN, Fagan SC, Ergul A (2013) Comparative analysis of the neurovascular injury and functional outcomes in experimental stroke models in diabetic Goto-Kakizaki rats. Brain Res 1541:106–114

    Article  CAS  PubMed  Google Scholar 

  • McCullough LD, Zeng Z, Li H, Landree LE, McFadden J, Ronnett GV (2005) Pharmacological inhibition of AMP-activated protein kinase provides neuroprotection in stroke. J Biol Chem 280(21):20493–20502

    Article  CAS  PubMed  Google Scholar 

  • Mielke JG, Taghibiglou C, Wang YT (2006) Endogenous insulin signaling protects cultured neurons from oxygen-glucose deprivation-induced cell death. Neuroscience 143:165–173

    Article  CAS  PubMed  Google Scholar 

  • Mohagheghi F, Khalaj L, Ahmadiani A, Rahmani B (2012) Gemfibrozil pretreatment affecting antioxidant defense system and inflammatory, but not Nrf-2 signaling pathways resulted in female neuroprotection and male neurotoxicity in the rat models of global cerebral ischemia-reperfusion. Neurotox Res 23:225–237

    Article  PubMed  Google Scholar 

  • Mohagheghi F, Ahmadiani A, Rahmani B, Moradi F, Romond N, Khalaj L (2013) Gemfibrozil pretreatment resulted in a sexually dimorphic outcome in the rat models of global cerebral ischemia-reperfusion via modulation of mitochondrial pro-survival and apoptotic cell death factors as well as MAPKs. J Mol Neurosci 50:379–393

    Article  CAS  PubMed  Google Scholar 

  • Mukherjee P, Mulrooney TJ, Marsh J, Blair D, Chiles TC, Seyfried TN (2008) Differential effects of energy stress on AMPK phosphorylation and apoptosis in experimental brain tumor and normal brain. Mol Cancer 7:37

    Article  PubMed Central  PubMed  Google Scholar 

  • Musi N, Hirshman MF, Nygren J, Svanfeldt M, Bavenholm P, Rooyackers O, Zhou G, Williamson JM, Ljunqvist O, Efendic S, Moller DE, Thorell A, Goodyear LJ (2002) Metformin increases AMP-activated protein kinase activity in skeletal muscle of subjects with type 2 diabetes. Diabetes 51:2074–2081

    Article  CAS  PubMed  Google Scholar 

  • Neurath KM, Keough MP, Mikkelsen T, Claffey KP (2006) AMP-dependent protein kinase alpha 2 isoform promotes hypoxia-induced VEGF expression in human glioblastoma. Glia 53:733–743

    Article  PubMed  Google Scholar 

  • Niimura M, Takagi N, Takagi K, Mizutani R, Ishihara N, Matsumoto K, Funakoshi H, Nakamura T, Takeo S (2006) Prevention of apoptosis-inducing factor translocation is a possible mechanism for protective effects of hepatocyte growth factor against neuronal cell death in the hippocampus after transient forebrain ischemia. J Cereb Blood Flow Metab 26:1354–1365

    Article  CAS  PubMed  Google Scholar 

  • Nita DA, Nita V, Spulber S, Moldovan M, Popa DP, Zagrean AM, Zagrean L (2001) Oxidative damage following cerebral ischemia depends on reperfusion - a biochemical study in rat. J Cell Mol Med 5:163–170

    Article  CAS  PubMed  Google Scholar 

  • Petersen JS, Andersen D, Muntzel MS, Diemer NH, Holstein-Rathlou NH (2001) Intracerebroventricular metformin attenuates salt-induced hypertension in spontaneously hypertensive rats. Am J Hypertens 14(11 Pt 1):1116–1122

    Article  CAS  PubMed  Google Scholar 

  • Petito CK, Torres-Munoz J, Roberts B, Olarte JP, Nowak TS Jr, Pulsinelli WA (1997) DNA fragmentation follows delayed neuronal death in CA1 neurons exposed to transient global ischemia in the rat. J Cereb Blood Flow Metab 17:967–976

    Article  CAS  PubMed  Google Scholar 

  • Pintana H, Apaijai N, Pratchayasakul W, Chattipakorn N, Chattipakorn SC (2012) Effects of metformin on learning and memory behaviors and brain mitochondrial functions in high fat diet induced insulin resistant rats. Life Sci 91:409–414

    Article  CAS  PubMed  Google Scholar 

  • Potter WB, O’Riordan KJ, Barnett D, Osting SM, Wagoner M, Burger C, Roopra A (2010) Metabolic regulation of neuronal plasticity by the energy sensor AMPK. PLoS One 5:e8996

    Article  PubMed Central  PubMed  Google Scholar 

  • Pulsinelli WA, Brierley JB (1979) A new model of bilateral hemispheric ischemia in the unanesthetized rat. Stroke 10:267

    Article  CAS  PubMed  Google Scholar 

  • Seo-Mayer PW, Thulin G, Zhang L, Alves DS, Ardito T, Kashgarian M, Caplan MJ (2011) Preactivation of AMPK by metformin may ameliorate the epithelial cell damage caused by renal ischemia. Am J Physiol Ren Physiol 301:F1346–F1357

    Article  CAS  Google Scholar 

  • Soraya H, Khorrami A, Garjani A, Maleki-Dizaji N (2013) Acute treatment with metformin improves cardiac function following isoproterenol induced myocardial infarction in rats. Pharmacol Rep 64:1476–1484

    Google Scholar 

  • Spasic MR, Callaerts P, Norga KK (2009) AMP-activated protein kinase (AMPK) molecular crossroad for metabolic control and survival of neurons. Neuroscientist 15:309–316

    Article  CAS  PubMed  Google Scholar 

  • Suwa M, Nakano H, Radak Z, Kumagai S (2010) Short-term adenosine monophosphate-activated protein kinase activator 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside treatment increases the sirtuin 1 protein expression in skeletal muscle. Metabolism 60:394–403

    Article  PubMed  Google Scholar 

  • Ullah I, Ullah N, Naseer MI, Lee HY, Kim MO (2012) Neuroprotection with metformin and thymoquinone against ethanol-induced apoptotic neurodegeneration in prenatal rat cortical neurons. BMC Neurosci 13:11

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Venna VR, Li J, Benashski SE, Tarabishy S, McCullough LD (2012) Preconditioning induces sustained neuroprotection by downregulation of adenosine 5′-monophosphate-activated protein kinase. Neuroscience 201:280–287

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Viollet B, Foretz M (2013) Revisiting the mechanisms of metformin action in the liver. Ann Endocrinol (Paris) 74:123–129

    Article  CAS  Google Scholar 

  • Vytla VS, Ochs RS (2013) Metformin increases mitochondrial energy formation in L6 muscle cell cultures. J Biol Chem 288:20369–20377

    Article  CAS  PubMed  Google Scholar 

  • Wang Q, Sun AY, Simonyi A, Kalogeris TJ, Miller DK, Sun GY, Korthuis RJ (2007) Ethanol preconditioning protects against ischemia/reperfusion-induced brain damage: role of NADPH oxidase-derived ROS. Free Radic Biol Med 43:1048–1060

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Yan W, Zhang H, Liu P, Wang H, Liu J, Gao C, Liu Y, Lian K, Yang L, Sun L, Guo Y, Zhang L, Dong L, Lau WB, Gao E, Gao F, Xiong L, Qu Y, Tao L (2013) Impaired mitochondrial biogenesis due to dysfunctional adiponectin-AMPK-PGC-1alpha signaling contributing to increased vulnerability in diabetic heart. Basic Res Cardiol 108(3):329

    Article  PubMed  Google Scholar 

  • Yeh CH, Chen TP, Wang YC, Lin YM, Fang SW (2011) AMP-activated protein kinase activation during cardioplegia-induced hypoxia/reoxygenation injury attenuates cardiomyocytic apoptosis via reduction of endoplasmic reticulum stress. Mediat Inflamm 2010:130636

    Google Scholar 

  • Zhang Q, Wu Y, Zhang P, Sha H, Jia J, Hu Y, Zhu J (2012) Exercise induces mitochondrial biogenesis after brain ischemia in rats. Neuroscience 205:10–17

    Article  CAS  PubMed  Google Scholar 

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Acknowledgment

We thank the research council of Alborz medical university for the funding of this project. We are also grateful to Neuroscience Research Center of Shahid Beheshti University for providing facilities of current study. Authors wish to thank Osvah Pharmaceutical Co. as well for providing us metformin as a generous gift. The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Department of Physiology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

    Ghorbangol Ashabi

  2. NeuroBiology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Fariba Khodagholi & Masoumeh Nasiri

  3. Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Fariba Khodagholi & Masoumeh Nasiri

  4. Medical School, Alborz University of Medical Sciences, Alborz, Iran

    Leila Khalaj & Mahdi Goudarzvand

  5. Physiology-Pharmacology Department, Medical School, Alborz University of Medical Sciences, P.O. Box: 3148/561 Eshteraki Ave., Baghestan Blvd., Karaj, Iran

    Leila Khalaj

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  1. Ghorbangol Ashabi
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  2. Fariba Khodagholi
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Correspondence to Leila Khalaj.

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Ashabi, G., Khodagholi, F., Khalaj, L. et al. Activation of AMP-activated protein kinase by metformin protects against global cerebral ischemia in male rats: Interference of AMPK/PGC-1α pathway. Metab Brain Dis 29, 47–58 (2014). https://doi.org/10.1007/s11011-013-9475-2

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