The immune response to sepsis involves a series of complex, highly integrated homeostatic responses that, if prolonged and excessive, can lead to organ dysfunction and death. Nitric oxide (NO) synthesis is upregulated by sepsis in many tissues and is an essential component of the host immune response. Nitric oxide synthesis can be beneficial and improve immune and organ function, but if synthesis is excessive and prolonged, NO can promote organ injury, tissue inflammation, and death. NO is produced in hepatocytes by the inducible nitric oxide synthase (iNOS) that is stimulated by cytokines and proinflammatory stimuli. Excessive NO from iNOS produces cellular dysfunction and hepatic injury. Glucagon and cyclic adenosine monophosphate (cAMP) regulate hepatic iNOS expression in vitro and in vivo, and by doing so, decrease NO-mediated hepatic injury. Our preliminary data demonstrate that insulin also down-regulates cytokine-induced iNOS expression. Both glucagon and insulin alter specific intracellular signaling pathways in hepatocytes, but the mechanisms involved in the regulation of hepatocyte function in sepsis by glucagon and insulin, and specifically the regulation of hepatocyte iNOS expression, have not been identified. In this proposal, we will determine the mechanisms responsible for the regulation of hepatocyte iNOS expression by glucagon and insulin.
In Aim I, we will continue our work in determining the mechanism for the glucagon and cAMP-induced inhibition of hepatocyte iNOS expression. We will focus on protein kinase A (PKA)-independent pathways induced by cAMP and evaluate the role of the guanine nucleotide exchange factor Epac and the role of calcium.
In Aim II, we will determine the mechanisms responsible for the inhibition of iNOS by insulin. By defining how these hormones regulate hepatocyte iNOS expression, we will provide a framework for understanding the basic pathophysiologic cellular events in shock and sepsis that may lead to novel cellular-based therapies for critically ill patients. Project Narrative Nitric oxide is synthesized in critically ill patients during septic shock, and when overproduced, can increase cellular dysfunction, tissue injury, and death. Glucagon and insulin primarily regulate blood glucose, which has become an important facet of the care of critically ill patients, but we have found that they also regulate hepatic nitric oxide production. We will determine the mechanisms responsible for the regulation of hepatocyte inducible nitric oxide synthase (iNOS) expression by glucagon and insulin. By defining these mechanisms, we will provide a framework for understanding the basic cellular events in shock and sepsis, which may lead to novel cellular-based therapies for critically ill patients.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK055664-10
Application #
8208065
Study Section
Surgery, Anesthesiology and Trauma Study Section (SAT)
Program Officer
Serrano, Jose
Project Start
2001-02-15
Project End
2013-02-28
Budget Start
2011-12-01
Budget End
2013-02-28
Support Year
10
Fiscal Year
2012
Total Cost
$308,241
Indirect Cost
$99,970
Name
University of Louisville
Department
Surgery
Type
Schools of Medicine
DUNS #
057588857
City
Louisville
State
KY
Country
United States
Zip Code
40292
Zhang, Baochun; Crankshaw, Will; Nesemeier, Ryan et al. (2015) Calcium-mediated signaling and calmodulin-dependent kinase regulate hepatocyte-inducible nitric oxide synthase expression. J Surg Res 193:795-801
Lakshmanan, Jaganathan; Zhang, Baochun; Nweze, Ikenna C et al. (2015) Glycogen synthase kinase 3 regulates IL-1β mediated iNOS expression in hepatocytes by down-regulating c-Jun. J Cell Biochem 116:133-41
Kimbrough, Charles W; Lakshmanan, Jaganathan; Matheson, Paul J et al. (2015) Resveratrol decreases nitric oxide production by hepatocytes during inflammation. Surgery 158:1095-101; discussion 1101
Zhang, Baochun; Nweze, Ikenna; Lakshmanan, Jaganathan et al. (2013) Activation of a cyclic amp-guanine exchange factor in hepatocytes decreases nitric oxide synthase expression. Shock 39:70-6
Nweze, Ikenna C; Smith, Jason W; Zhang, Baochun et al. (2012) 17β-Estradiol attenuates cytokine-induced nitric oxide production in rat hepatocyte. J Trauma Acute Care Surg 73:408-12
Harbrecht, Brian G; Nweze, Ikenna; Smith, Jason W et al. (2012) Insulin inhibits hepatocyte iNOS expression induced by cytokines by an Akt-dependent mechanism. Am J Physiol Gastrointest Liver Physiol 302:G116-22
Miller, Keith R; Lawson, Christy M; Smith, Vance L et al. (2011) Carbohydrate provision in the era of tight glucose control. Curr Gastroenterol Rep 13:388-94
Zhang, Baochun; Li, Suping; Harbrecht, Brian G (2011) Akt-mediated signaling is induced by cytokines and cyclic adenosine monophosphate and suppresses hepatocyte inducible nitric oxide synthase expression independent of MAPK P44/42. Biochim Biophys Acta 1813:73-9
Smith, Jason W; Garrison, R Neal; Matheson, Paul J et al. (2010) Direct peritoneal resuscitation accelerates primary abdominal wall closure after damage control surgery. J Am Coll Surg 210:658-64, 664-7
Hong, Guiying; Zhang, Baochun; Harbrecht, Brian G (2010) Cyclic AMP inhibits IL-1beta plus IFNgamma-induced NF-kappaB translocation in hepatocytes by a PKA independent mechanism. J Surg Res 159:565-71

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