AMPK activation protects astrocytes from hypoxia‑induced cell death

Barialai,Leli; Strecker,Maja   I.; Luger,Anna‑Luisa; Jäger,Manuel; Bruns,Ines; Sittig,Alina  C.M.; Mildenberger,Iris   C.; Heller,Sonja  M.; Delaidelli,Alberto; Lorenz,Nadja   I.; Voss,Martin; Ronellenfitsch,Michael   W.; Steinbach,Joachim   P.; Burger,Michael   C.

doi:10.3892/ijmm.2020.4528

AMPK activation protects astrocytes from hypoxia‑induced cell death

Authors:
- Leli Barialai
- Maja I. Strecker
- Anna‑Luisa Luger
- Manuel Jäger
- Ines Bruns
- Alina C.M. Sittig
- Iris C. Mildenberger
- Sonja M. Heller
- Alberto Delaidelli
- Nadja I. Lorenz
- Martin Voss
- Michael W. Ronellenfitsch
- Joachim P. Steinbach
- Michael C. Burger
View Affiliations

Affiliations: Dr. Senckenberg Institute of Neurooncology, University Hospital Frankfurt, Goethe University, D-60528 Frankfurt am Main, Germany, Department of Dermatology, Venerology and Allergology, University Hospital Frankfurt, Goethe University, D-60590 Frankfurt am Main
Published online on: March 5, 2020 https://doi.org/10.3892/ijmm.2020.4528
Pages: 1385-1396
Copyright: © Barialai et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Adenosine monophosphate (AMP)‑activated protein kinase (AMPK) is a major cellular energy sensor that is activated by an increase in the AMP/adenosine triphosphate (ATP) ratio. This causes the initiation of adaptive cellular programs, leading to the inhibition of anabolic pathways and increasing ATP synthesis. AMPK indirectly inhibits mammalian target of rapamycin (mTOR) complex 1 (mTORC1), a serine/threonine kinase and central regulator of cell growth and metabolism, which integrates various growth inhibitory signals, such as the depletion of glucose, amino acids, ATP and oxygen. While neuroprotective approaches by definition focus on neurons, that are more sensitive under cell stress conditions, astrocytes play an important role in the cerebral energy homeostasis during ischemia. Therefore, the protection of astrocytic cells or other glial cells may contribute to the preservation of neuronal integrity and function. In the present study, it was thus hypothesized that a preventive induction of energy deprivation‑activated signaling pathways via AMPK may protect astrocytes from hypoxia and glucose deprivation. Hypoxia‑induced cell death was measured in a paradigm of hypoxia and partial glucose deprivation in vitro in the immortalized human astrocytic cell line SVG. Both the glycolysis inhibitor 2‑deoxy‑d‑glucose (2DG) and the AMPK activator A‑769662 induced the phosphorylation of AMPK, resulting in mTORC1 inhibition, as evidenced by a decrease in the phosphorylation of the target ribosomal protein S6 (RPS6). Treatment with both 2DG and A‑769662 also decreased glucose consumption and lactate production. Furthermore, A‑769662, but not 2DG induced an increase in oxygen consumption, possibly indicating a more efficient glucose utilization through oxidative phosphorylation. Hypoxia‑induced cell death was profoundly reduced by treatment with 2DG or A‑769662. On the whole, the findings of the present study demonstrate, that AMPK activation via 2DG or A‑769662 protects astrocytes under hypoxic and glucose‑depleted conditions.

View Figures

View References

1	Hardie DG, Ross FA and Hawley SA: AMPK: A nutrient and energy sensor that maintains energy homeostasis. Nat Rev Mol Cell Biol. 13:251–262. 2012. View Article : Google Scholar : PubMed/NCBI
2	Mitsudomi T and Yatabe Y: Epidermal growth factor receptor in relation to tumor development: EGFR gene and cancer. FEBS J. 277:301–308. 2010. View Article : Google Scholar
3	Wullschleger S, Loewith R and Hall MN: TOR signaling in growth and metabolism. Cell. 124:471–484. 2006. View Article : Google Scholar : PubMed/NCBI
4	Hardie DG and Pan DA: Regulation of fatty acid synthesis and oxidation by the AMP-activated protein kinase. Biochem Soc Trans. 30:1064–1070. 2002. View Article : Google Scholar : PubMed/NCBI
5	Goransson O, McBride A, Hawley SA, Ross FA, Shpiro N, Foretz M, Viollet B, Hardie DG and Sakamoto K: Mechanism of action of A-769662, a valuable tool for activation of AMP-activated protein kinase. J Biol Chem. 282:32549–32560. 2007. View Article : Google Scholar : PubMed/NCBI
6	Hartel I, Ronellenfitsch M, Wanka C, Wolking S, Steinbach JP and Rieger J: Activation of AMP-activated kinase modulates sensitivity of glioma cells against epidermal growth factor receptor inhibition. Int J Oncol. 49:173–180. 2016. View Article : Google Scholar : PubMed/NCBI
7	Strickland M and Stoll EA: Metabolic reprogramming in glioma. Front Cell Dev Biol. 5:432017. View Article : Google Scholar : PubMed/NCBI
8	Woodward GE and Hudson MT: The effect of 2-desoxy-D-glucose on glycolysis and respiration of tumor and normal tissues. Cancer Res. 14:599–605. 1954.PubMed/NCBI
9	Wick AN, Drury DR, Nakada HI and Wolfe JB: Localization of the primary metabolic block produced by 2-deoxyglucose. J Biol Chem. 224:963–969. 1957.PubMed/NCBI
10	Yang C, Ko B, Hensley CT, Jiang L, Wasti AT, Kim J, Sudderth J, Calvaruso MA, Lumata L, Mitsche M, et al: Glutamine oxidation maintains the TCA cycle and cell survival during impaired mitochondrial pyruvate transport. Mol Cell. 56:414–424. 2014. View Article : Google Scholar : PubMed/NCBI
11	Sellers K, Fox MP, Bousamra M II, Slone SP, Higashi RM, Miller DM, Wang Y, Yan J, Yuneva MO, Deshpande R, et al: Pyruvate carboxylase is critical for non-small-cell lung cancer proliferation. J Clin Invest. 125:687–698. 2015. View Article : Google Scholar : PubMed/NCBI
12	Csibi A, Fendt SM, Li C, Poulogiannis G, Choo AY, Chapski DJ, Jeong SM, Dempsey JM, Parkhitko A, Morrison T, et al: The mTORC1 pathway stimulates glutamine metabolism and cell proliferation by repressing SIRT4. Cell. 153:840–854. 2013. View Article : Google Scholar : PubMed/NCBI
13	Becerra-Calixto A and Cardona-Gomez GP: The role of astrocytes in neuroprotection after brain stroke: Potential in cell therapy. Front Mol Neurosci. 10:882017. View Article : Google Scholar : PubMed/NCBI
14	Heiss WD: The concept of the penumbra: Can it be translated to stroke management? Int J Stroke. 5:290–295. 2010. View Article : Google Scholar : PubMed/NCBI
15	Candelario-Jalil E: Injury and repair mechanisms in ischemic stroke: Considerations for the development of novel neurotherapeutics. Curr Opin Investig Drugs. 10:644–654. 2009.PubMed/NCBI
16	Khaladj N, Peterss S, Oetjen P, von Wasielewski R, Hauschild G, Karck M, Haverich A and Hagl C: Hypothermic circulatory arrest with moderate, deep or profound hypothermic selective antegrade cerebral perfusion: Which temperature provides best brain protection? Eur J Cardiothorac Surg. 30:492–498. 2006. View Article : Google Scholar : PubMed/NCBI
17	Pacini D, Leone A, Di Marco L, Marsilli D, Sobaih F, Turci S, Masieri V and Di Bartolomeo R: Antegrade selective cerebral perfusion in thoracic aorta surgery: Safety of moderate hypothermia. Eur J Cardiothorac Surg. 31:618–622. 2007. View Article : Google Scholar : PubMed/NCBI
18	Abbott NJ, Ronnback L and Hansson E: Astrocyte-endothelial interactions at the blood-brain barrier. Nat Rev Neurosci. 7:41–53. 2006. View Article : Google Scholar
19	Newman EA: Glial cell regulation of neuronal activity and blood flow in the retina by release of gliotransmitters. Philos Trans R Soc Lond B Biol Sci. 370:2015. View Article : Google Scholar : PubMed/NCBI
20	Kitchen P, Day RE, Taylor LH, Salman MM, Bill RM, Conner MT and Conner AC: Identification and molecular mechanisms of the rapid tonicity-induced relocalization of the aquaporin 4 channel. J Biol Chem. 290:16873–16881. 2015. View Article : Google Scholar : PubMed/NCBI
21	Kaczor P, Rakus D and Mozrzymas JW: Neuron-astrocyte interaction enhance GABAergic synaptic transmission in a manner dependent on key metabolic enzymes. Front Cell Neurosci. 9:1202015. View Article : Google Scholar : PubMed/NCBI
22	Xie Z, Ding SQ and Shen YF: Silibinin activates AMP-activated protein kinase to protect neuronal cells from oxygen and glucose deprivation-re-oxygenation. Biochem Biophys Res Commun. 454:313–319. 2014. View Article : Google Scholar : PubMed/NCBI
23	Major EO, Miller AE, Mourrain P, Traub RG, de Widt E and Sever J: Establishment of a line of human fetal glial cells that supports JC virus multiplication. Proc Natl Acad Sci USA. 82:1257–1261. 1985. View Article : Google Scholar : PubMed/NCBI
24	Bachis A, Major EO and Mocchetti I: Brain-derived neurotrophic factor inhibits human immunodeficiency virus-1/gp120-mediated cerebellar granule cell death by preventing gp120 internalization. J Neurosci. 23:5715–5722. 2003. View Article : Google Scholar : PubMed/NCBI
25	Steinbach JP, Wolburg H, Klumpp A, Probst H and Weller M: Hypoxia-induced cell death in human malignant glioma cells: Energy deprivation promotes decoupling of mitochondrial cytochrome c release from caspase processing and necrotic cell death. Cell Death Differ. 10:823–832. 2003. View Article : Google Scholar : PubMed/NCBI
26	Steinbach JP, Klumpp A, Wolburg H and Weller M: Inhibition of epidermal growth factor receptor signaling protects human malignant glioma cells from hypoxia-induced cell death. Cancer Res. 64:1575–1578. 2004. View Article : Google Scholar : PubMed/NCBI
27	Thiepold AL, Lorenz NI, Foltyn M, Engel AL, Divé I, Urban H, Heller S, Bruns I, Hofmann U, Dröse S, et al: Mammalian target of rapamycin complex 1 activation sensitizes human glioma cells to hypoxia-induced cell death. Brain. 140:2623–2638. 2017. View Article : Google Scholar : PubMed/NCBI
28	Ronellenfitsch MW, Brucker DP, Burger MC, Wolking S, Tritschler F, Rieger J, Wick W, Weller M and Steinbach JP: Antagonism of the mammalian target of rapamycin selectively mediates metabolic effects of epidermal growth factor receptor inhibition and protects human malignant glioma cells from hypoxia-induced cell death. Brain. 132:1509–1522. 2009. View Article : Google Scholar : PubMed/NCBI
29	Wise DR, Ward PS, Shay JE, Cross JR, Gruber JJ, Sachdeva UM, Platt JM, DeMatteo RG, Simon MC and Thompson CB: Hypoxia promotes isocitrate dehydrogenase-dependent carboxylation of α-ketoglutarate to citrate to support cell growth and viability. Proc Natl Acad Sci USA. 108:19611–19616. 2011. View Article : Google Scholar
30	Gabryel B, Kost A, Kasprowska D, Liber S, Machnik G, Wiaderkiewicz R and Łabuzek K: AMP-activated protein kinase is involved in induction of protective autophagy in astrocytes exposed to oxygen-glucose deprivation. Cell Biol Int. 38:1086–1097. 2014. View Article : Google Scholar : PubMed/NCBI
31	Gabryel B and Liber S: Metformin limits apoptosis in primary rat cortical astrocytes subjected to oxygen and glucose deprivation. Folia Neuropathol. 56:328–336. 2018. View Article : Google Scholar
32	Guo X, Jiang Q, Tuccitto A, Chan D, Alqawlaq S, Won GJ and Sivak JM: The AMPK-PGC-1α signaling axis regulates the astrocyte glutathione system to protect against oxidative and metabolic injury. Neurobiol Dis. 113:59–69. 2018. View Article : Google Scholar : PubMed/NCBI
33	Viollet B, Guigas B, Leclerc J, Hébrard S, Lantier L, Mounier R, Andreelli F and Foretz M: AMP-activated protein kinase in the regulation of hepatic energy metabolism: From physiology to therapeutic perspectives. Acta Physiol (Oxf). 196:81–98. 2009. View Article : Google Scholar
34	Dasgupta B and Seibel W: Compound c/dorsomorphin: Its use and misuse as an AMPK inhibitor. Methods Mol Biol. 1732:195–202. 2018. View Article : Google Scholar : PubMed/NCBI
35	Shin SY, Kim TH, Wu H, Choi YH and Kim SG: SIRT1 activation by methylene blue, a repurposed drug, leads to AMPK-mediated inhibition of steatosis and steatohepatitis. Eur J Pharmacol. 727:115–124. 2014. View Article : Google Scholar : PubMed/NCBI
36	Wu L, Zhang L, Li B, Jiang H, Duan Y, Xie Z, Shuai L and Li J and Li J: AMP-activated protein kinase (AMPK) regulates energy metabolism through modulating thermogenesis in adipose tissue. Front Physiol. 9:1222018. View Article : Google Scholar : PubMed/NCBI
37	Vlachaki Walker JM, Robb JL, Cruz AM, Malhi A, Weightman Potter PG, Ashford MLJ, McCrimmon RJ, Ellacott KLJ and Beall C: AMP-activated protein kinase (AMPK) activator A-769662 increases intracellular calcium and ATP release from astrocytes in an AMPK-independent manner. Diabetes Obes Metab. 19:997–1005. 2017. View Article : Google Scholar : PubMed/NCBI
38	Zhao X, Zhou KS, Li ZH, Nan W, Wang J, Xia YY and Zhang HH: Knockdown of Ski decreased the reactive astrocytes proliferation in vitro induced by oxygen-glucose deprivation/reoxygenation. J Cell Biochem. 119:4548–4558. 2018. View Article : Google Scholar
39	He M, Shi X, Yang M, Yang T, Li T and Chen J: Mesenchymal stem cells-derived IL-6 activates AMPK/mTOR signaling to inhibit the proliferation of reactive astrocytes induced by hypoxic-ischemic brain damage. Exp Neurol. 311:15–32. 2019. View Article : Google Scholar
40	Lin L, Bivard A and Parsons MW: Perfusion patterns of ischemic stroke on computed tomography perfusion. J Stroke. 15:164–173. 2013. View Article : Google Scholar
41	Lee DH, Kang DW, Ahn JS, Choi CG, Kim SJ and Suh DC: Imaging of the ischemic penumbra in acute stroke. Korean J Radiol. 6:64–74. 2005. View Article : Google Scholar : PubMed/NCBI