Molecular Medicine

, Volume 21, Issue 1, pp 937–950 | Cite as

AMP-Activated Protein Kinase and Glycogen Synthase Kinase 3β Modulate the Severity of Sepsis-induced Lung injury

  • Zhongyu Liu
  • Nathaniel Bone
  • Shaoning Jiang
  • Dae Won Park
  • Jean-Marc Tadie
  • Jessy Deshane
  • Cilina Ann Rodriguez
  • Jean-Francois Pittet
  • Edward Abraham
  • Jaroslaw W. Zmijewski
Research Article


Alterations in metabolic and bioenergetic homeostasis contribute to sepsis-mediated organ injury. However, how AMP-activated protein kinase (AMPK), a major sensor and regulator of energy expenditure and production, affects development of organ injury and loss of innate capacity during polymicrobial sepsis remains unclear. In the present experiments, we found that cross-talk between the AMPK and GSK3β signaling pathways controls chemotaxis and the ability of neutrophils and macrophages to kill bacteria ex vivo. In mice with polymicrobial abdominal sepsis or more severe sepsis induced by the combination of hemorrhage and intraabdominal infection, administration of the AMPK activator metformin or the GSK3β inhibitor SB216763 reduced the severity of acute lung injury (ALI). Improved survival in metformin-treated septic mice was correlated with preservation of mitochondrial complex V (ATP synthase) function and increased amounts of ETC complex III and IV. Although immunosuppression is a consequence of sepsis, metformin effectively increased innate immune capacity to eradicate P. aeruginosa in the lungs of septic mice. We also found that AMPK activation diminished accumulation of the immunosuppressive transcriptional factor HIF-1α as well as the development of endotoxin tolerance in LPS-treated macrophages. Furthermore, AMPK-dependent preservation of mitochondrial membrane potential also prevented LPS-mediated dysfunction of neutrophil chemotaxis. These results indicate that AMPK activation reduces the severity of polymicrobial sepsis-induced lung injury and prevents the development of sepsis-associated immunosuppression.



We thank Ken Inoki from the University of Michigan for the anti-phospho-Thr479-AMPK antibody. Funding was provided by National Institutes of Health Grant HL107585 to JW Zmijewski.

Supplementary material

10020_2015_2101937_MOESM1_ESM.pdf (364 kb)
Supplementary material, approximately 364 KB.


  1. 1.
    Deutschman CS, Tracey KJ. (2014) Sepsis: current dogma and new perspectives. Immunity. 40:463–75.CrossRefGoogle Scholar
  2. 2.
    Angus DC, van der Poll T. (2013) Severe sepsis and septic shock. N. Engl. J. Med. 369:2063.CrossRefGoogle Scholar
  3. 3.
    Hotchkiss RS, Monneret G, Payen D. (2013) Immunosuppression in sepsis: a novel understanding of the disorder and a new therapeutic approach. Lancet Infect. Dis. 13:260–8.CrossRefGoogle Scholar
  4. 4.
    Lord JM, et al. (2014) The systemic immune response to trauma: an overview of pathophysiology and treatment. Lancet. 384:1455–65.CrossRefGoogle Scholar
  5. 5.
    Wheeler AP, Bernard GR. (2007) Acute lung injury and the acute respiratory distress syndrome: a clinical review. Lancet. 369:1553–64.CrossRefGoogle Scholar
  6. 6.
    Force ADT, et al. (2012) Acute respiratory distress syndrome: the Berlin Definition. JAMA. 307:2526–33.Google Scholar
  7. 7.
    Sheu CC, et al. (2010) Clinical characteristics and outcomes of sepsis-related vs non-sepsis-related ARDS. Chest. 138:559–67.CrossRefGoogle Scholar
  8. 8.
    Fink MP, Warren HS. (2014) Strategies to improve drug development for sepsis. Nat. Rev. Drug. Discov. 13:741–58.CrossRefGoogle Scholar
  9. 9.
    Standiford TJ, Ward PA. (2016) Therapeutic targeting of acute lung injury and acute respiratory distress syndrome. Transl. Res. 167:183–91.CrossRefGoogle Scholar
  10. 10.
    Flierl MA, et al. (2008) Adverse functions of IL-17A in experimental sepsis. FASEB J. 22:2198–205.CrossRefGoogle Scholar
  11. 11.
    Bosmann M, Ward PA. (2012) Therapeutic potential of targeting IL-17 and IL-23 in sepsis. Clin. Transl. Med. 1:4.CrossRefGoogle Scholar
  12. 12.
    Mariathasan S, et al. (2004) Differential activation of the inflammasome by caspase-1 adaptors ASC and Ipaf. Nature. 430:213–8.CrossRefGoogle Scholar
  13. 13.
    Brown KA, et al. (2006) Neutrophils in development of multiple organ failure in sepsis. Lancet. 368:157–69.CrossRefGoogle Scholar
  14. 14.
    Kolaczkowska E, Kubes P. (2013) Neutrophil recruitment and function in health and inflammation. Nat. Rev. Immunol. 13:159–75.CrossRefGoogle Scholar
  15. 15.
    Abraham E. (2003) Neutrophils and acute lung injury. Crit. Care Med. 31:S195–9.CrossRefGoogle Scholar
  16. 16.
    Hotchkiss RS, Monneret G, Payen D. (2013) Sepsis-induced immunosuppression: from cellular dysfunctions to immunotherapy. Nat. Rev. Immunol. 13:862–74.CrossRefGoogle Scholar
  17. 17.
    Gentile LF, et al. (2012) Persistent inflammation and immunosuppression: a common syndrome and new horizon for surgical intensive care. J. Trauma Acute Care Surg. 72:1491–501.CrossRefGoogle Scholar
  18. 18.
    Singer M. (2014) The role of mitochondrial dysfunction in sepsis-induced multi-organ failure. Virulence. 5:66–72.CrossRefGoogle Scholar
  19. 19.
    Brealey D, et al. (2002) Association between mitochondrial dysfunction and severity and outcome of septic shock. Lancet. 360:219–23.CrossRefGoogle Scholar
  20. 20.
    Singer M. (2007) Mitochondrial function in sepsis: acute phase versus multiple organ failure. Crit. Care Med. 35:S441–8.CrossRefGoogle Scholar
  21. 21.
    Srivastava A, et al. (2015) MKK3 deletion improves mitochondrial quality. Free Radic. Biol. Med. 87:373–84.CrossRefGoogle Scholar
  22. 22.
    Matthay MA. (2015) Therapeutic potential of mesenchymal stromal cells for acute respiratory distress syndrome. Ann. Am. Thorac. Soc. 12 Suppl 1:S54–7.CrossRefGoogle Scholar
  23. 23.
    Rocha M, Herance R, Rovira S, Hernandez-Mijares A, Victor VM. (2012) Mitochondrial dysfunction and antioxidant therapy in sepsis. Infect. Disord. Drug Targets. 12:161–78.CrossRefGoogle Scholar
  24. 24.
    Hardie DG, Ross FA, Hawley SA. (2012) AMPK: a nutrient and energy sensor that maintains energy homeostasis. Nat. Rev. Mol. Cell. Biol. 13:251–62.CrossRefGoogle Scholar
  25. 25.
    Eid AA, Lee DY, Roman LJ, Khazim K, Gorin Y. (2013) Sestrin 2 and AMPK connect hyperglycemia to Nox4-dependent endothelial nitric oxide synthase uncoupling and matrix protein expression. Mol. Cell. Biol. 33:3439–60.CrossRefGoogle Scholar
  26. 26.
    Colombo SL, Moncada S. (2009) AMPKalpha1 regulates the antioxidant status of vascular endothelial cells. Biochem. J. 421:163–9.CrossRefGoogle Scholar
  27. 27.
    Chen L, et al. (2009) Structural insight into the autoinhibition mechanism of AMP-activated protein kinase. Nature. 459:1146–9.CrossRefGoogle Scholar
  28. 28.
    Woods A, et al. (2003) Identification of phosphorylation sites in AMP-activated protein kinase (AMPK) for upstream AMPK kinases and study of their roles by site-directed mutagenesis. J. Biol. Chem. 278:28434–42.CrossRefGoogle Scholar
  29. 29.
    Martin-Montalvo A, et al. (2013) Metformin improves healthspan and lifespan in mice. Nat. Commun. 4:2192.CrossRefGoogle Scholar
  30. 30.
    Bannister CA, et al. (2014) Can people with type 2 diabetes live longer than those without? A comparison of mortality in people initiated with metformin or sulphonylurea monotherapy and matched, non-diabetic controls. Diabetes Obes. Metab. 16:1165–73.CrossRefGoogle Scholar
  31. 31.
    Zhao X, et al. (2008) Activation of AMPK attenuates neutrophil proinflammatory activity and decreases the severity of acute lung injury. Am. J. Physiol. Lung Cell. Mol. Physiol. 295:L497–504.CrossRefGoogle Scholar
  32. 32.
    Zmijewski JW, et al. (2008) Mitochondrial respiratory complex I regulates neutrophil activation and severity of lung injury. Am. J. Respir. Crit. Care Med. 178:168–79.CrossRefGoogle Scholar
  33. 33.
    Jian MY, Alexeyev MF, Wolkowicz PE, Zmijewski JW, Creighton JR. (2013) Metformin-stimulated AMPK-α1 promotes microvascular repair in acute lung injury. Am. J. Physiol. Lung Cell. Mol. Physiol. 305:L844–55.CrossRefGoogle Scholar
  34. 34.
    Jiang S, et al. (2014) Human resistin promotes neu-trophil proinflammatory activation and neutrophil extracellular trap formation and increases severity of acute lung injury. J. Immunol. 192:4795–803.CrossRefGoogle Scholar
  35. 35.
    Tadie JM, et al. (2012) Toll-like receptor 4 engagement inhibits adenosine 5′-monophosphate-activated protein kinase activation through a high mobility group box 1 protein-dependent mechanism. Mol. Med. 18:659–68.CrossRefGoogle Scholar
  36. 36.
    Xing J, et al. (2013) Inhibition of AMP-activated protein kinase accentuates lipopolysaccharide-induced lung endothelial barrier dysfunction and lung injury in vivo. Am. J. Pathol. 182:1021–30.CrossRefGoogle Scholar
  37. 37.
    Meares GP, Qin H, Liu Y, Holdbrooks AT, Benveniste EN. (2013) AMP-activated protein kinase restricts IFN-gamma signaling. J. Immunol. 190:372–80.CrossRefGoogle Scholar
  38. 38.
    Park DW, et al. (2014) GSK3β-dependent inhibition of AMPK potentiates activation of neutrophils and macrophages and enhances severity of acute lung injury. Am. J. Physiol. Lung Cell. Mol. Physiol. 307:L735–45.CrossRefGoogle Scholar
  39. 39.
    Suzuki T, et al. (2013) Inhibition of AMPK catabolic action by GSK3. Mol. Cell 50:407–19.CrossRefGoogle Scholar
  40. 40.
    Jiang S, et al. (2013) Mitochondria and AMP-activated protein kinase-dependent mechanism of efferocytosis. J. Biol. Chem. 288:26013–26.CrossRefGoogle Scholar
  41. 41.
    Rittirsch D, Huber-Lang MS, Flierl MA, Ward PA. (2009) Immunodesign of experimental sepsis by cecal ligation and puncture. Nat. Protoc. 4:31–6.CrossRefGoogle Scholar
  42. 42.
    Rodriguez C, et al. (2009) Sodium nitrite therapy attenuates the hypertensive effects of HBOC-201 via nitrite reduction. Biochem. J. 422:423–32.CrossRefGoogle Scholar
  43. 43.
    Zmijewski JW, et al. (2009) Antiinflammatory effects of hydrogen peroxide in neutrophil activation and acute lung injury. Am. J. Respir. Crit. Care Med. 179:694–704.CrossRefGoogle Scholar
  44. 44.
    Latz E, Xiao TS, Stutz A. (2013) Activation and regulation of the inflammasomes. Nat. Rev. Immunol. 13:397–411.CrossRefGoogle Scholar
  45. 45.
    Ayala A, et al. (2002) Shock-induced neutrophil mediated priming for acute lung injury in mice: divergent effects of TLR-4 and TLR-4/FasL deficiency. Am. J. Pathol. 161:2283–94.CrossRefGoogle Scholar
  46. 46.
    LaRue KE, McCall CE. (1994) A labile transcriptional repressor modulates endotoxin tolerance. J. Exp. Med. 180:2269–75.CrossRefGoogle Scholar
  47. 47.
    McClure C, Brudecki L, Yao ZQ, McCall CE, El Gazzar M. (2015) Processing body formation limits proinflammatory cytokine synthesis in endotoxin-tolerant monocytes and murine septic macrophages. J. Innate Immun. 7:572–83.CrossRefGoogle Scholar
  48. 48.
    Sag D, Carling D, Stout RD, Suttles J. (2008) Adenosine 5′-monophosphate-activated protein kinase promotes macrophage polarization to an anti-inflammatory functional phenotype. J. Immunol. 181:8633–41.CrossRefGoogle Scholar
  49. 49.
    Jeong HW, et al. (2009) Berberine suppresses proinflammatory responses through AMPK activation in macrophages. Am. J. Physiol. Endocrinol. Metab. 296:E955–64.CrossRefGoogle Scholar
  50. 50.
    Shalova IN, et al. (2015) Human monocytes undergo functional re-programming during sepsis mediated by hypoxia-inducible factor-1α. Immunity. 42:484–98.CrossRefGoogle Scholar
  51. 51.
    van der Poll T, Opal SM. (2008) Host-pathogen interactions in sepsis. Lancet Infect. Dis. 8:32–43.CrossRefGoogle Scholar
  52. 52.
    Tavares-Murta BM, et al. (2002) Failure of neutrophil chemotactic function in septic patients. Crit. Care Med. 30:1056–61.CrossRefGoogle Scholar
  53. 53.
    Fossati G, et al. (2003) The mitochondrial network of human neutrophils: role in chemotaxis, phagocytosis, respiratory burst activation, and commitment to apoptosis. J. Immunol. 170: 1964–72.CrossRefGoogle Scholar
  54. 54.
    Barbier F, Andremont A, Wolff M, Bouadma L. (2013) Hospital-acquired pneumonia and ventilator-associated pneumonia: recent advances in epidemiology and management. Curr. Opin. Pulm. Med. 19:216–28.CrossRefGoogle Scholar
  55. 55.
    Martin M, Rehani K, Jope RS, Michalek SM. (2005) Toll-like receptor-mediated cytokine production is differentially regulated by glycogen synthase kinase 3. Nat. Immunol. 6:777–84.CrossRefGoogle Scholar
  56. 56.
    Rocha J, et al. (2015) Inhibition of glycogen synthase kinase-3β attenuates organ injury and dysfunction associated with liver ischemia-reperfusion and thermal injury in the rat. Shock. 43:369–78.CrossRefGoogle Scholar
  57. 57.
    Li H, et al. (2013) NF-κB inhibition after cecal ligation and puncture reduces sepsis-associated lung injury without altering bacterial host defense. Mediators Inflamm. 2013:503213.PubMedPubMedCentralGoogle Scholar
  58. 58.
    Park DW, et al. (2013) Activation of AMPK enhances neutrophil chemotaxis and bacterial killing. Mol. Med. 19:387–98.CrossRefGoogle Scholar
  59. 59.
    Bae HB, et al. (2011) AMP-activated protein kinase enhances the phagocytic ability of macrophages and neutrophils. FASEB J. 25: 4358–68.CrossRefGoogle Scholar
  60. 60.
    Mounier R, et al. (2013) AMPKα1 regulates macrophage skewing at the time of resolution of inflammation during skeletal muscle regeneration. Cell Metab. 18:251–64.CrossRefGoogle Scholar
  61. 61.
    Labuzek K, Liber S, Gabryel B, Adamczyk J, Okopien B. Metformin increases phagocytosis and acidifies lysosomal/endosomal compartments in AMPK-dependent manner in rat primary microglia. Naunyn Schmiedebergs Arch. Pharmacol. 381:171–86.Google Scholar
  62. 62.
    Zolfaghari PS, et al. (2015) Skeletal muscle dysfunction is associated with derangements in mitochondrial bioenergetics (but not UCP3) in a rodent model of sepsis. Am. J. Physiol. Endocrinol. Metab. 308:E713–25.CrossRefGoogle Scholar
  63. 63.
    Japiassu AM, et al. (2011) Bioenergetic failure of human peripheral blood monocytes in patients with septic shock is mediated by reduced F1Fo adenosine-5′-triphosphate synthase activity. Crit. Care Med. 39:1056–63.CrossRefGoogle Scholar
  64. 64.
    Mulchandani N, et al. (2015) Stimulation of brain AMP-activated protein kinase attenuates inflammation and acute lung injury in sepsis. Mol. Med. 21:637–44.CrossRefGoogle Scholar
  65. 65.
    Mannam P, et al. (2014) MKK3 regulates mitochondrial biogenesis and mitophagy in sepsis-induced lung injury. Am. J. Physiol. Lung Cell. Mol. Physiol. 306:L604–19.CrossRefGoogle Scholar
  66. 66.
    Ward PA. (2011) Immunosuppression in sepsis. JAMA. 306:2618–9.CrossRefGoogle Scholar
  67. 67.
    Faubert B, et al. (2013) AMPK is a negative regulator of the Warburg effect and suppresses tumor growth in vivo. Cell Metab. 17:113–24.CrossRefGoogle Scholar
  68. 68.
    Treins C, Murdaca J, Van Obberghen E, Giorgetti-Peraldi S. (2006) AMPK activation inhibits the expression of HIF-1alpha induced by insulin and IGF-1. Biochem. Biophys. Res. Commun. 342:1197–202.CrossRefGoogle Scholar
  69. 69.
    Metformin in Longevity Study (MILES) [Internet]. (2016) [updated 2015 Dec 8; cited 2016 Jan 29]. Available from: identifier: NCT02432287.

Copyright information

© The Author(s) 2015

Authors and Affiliations

  • Zhongyu Liu
    • 1
  • Nathaniel Bone
    • 1
  • Shaoning Jiang
    • 1
  • Dae Won Park
    • 1
  • Jean-Marc Tadie
    • 1
  • Jessy Deshane
    • 1
  • Cilina Ann Rodriguez
    • 2
  • Jean-Francois Pittet
    • 2
  • Edward Abraham
    • 3
  • Jaroslaw W. Zmijewski
    • 1
  1. 1.Department of Medicine, Division of Pulmonary, Allergy & Critical Care MedicineUniversity of Alabama at Birmingham, School of MedicineBirminghamUSA
  2. 2.Department of AnesthesiologyUniversity of Alabama at BirminghamBirminghamUSA
  3. 3.Office of the DeanWake Forest University School of MedicineWinston-SalemUSA

Personalised recommendations