Molecular Medicine

, Volume 14, Issue 1–2, pp 11–19 | Cite as

Insulin Decreases Inflammatory Signal Transcription Factor Expression in Primary Human Liver Cells after LPS Challenge

  • Marc G. Jeschke
  • Dagmar Klein
  • Wolfgang E. Thasler
  • Ulrich Bolder
  • Hans-Jürgen Schlitt
  • Karl-Walter Jauch
  • Thomas S. Weiss
Research Article


Hepatic homeostasis is essential for survival in critically ill and burned patients. Insulin administration improves survival and decreases infections in these patients. To determine the molecular mechanisms, the aim of the present study was to establish a stress model using primary human hepatocytes (PHHs) and to study the effects of insulin on the hepatic inflammatory signaling cascade. Liver tissue was obtained from general surgical patients, and PHHs were isolated and maintained in culture. Primary hepatocyte cultures were challenged with various doses of lipopolysaccharide (LPS), and the inflammatory signal transcription cascade was determined by real-time PCR. In subsequent experiments, primary hepatocyte cultures were challenged with LPS and insulin was added in various doses. Glucose was determined by colorimetric assays. PHHs treated with 100 µg/mL LPS showed a profound inflammatory reaction with increased expression of interleukin (IL)-6, IL-10, IL-1β, tumor necrosis factor (TNF), and signal transducer and activator of transcription 5 (STAT-5). Insulin at 10 IU/mL significantly decreased IL-6, TNF, and IL-1β at pretranslational levels, an effect associated with decreased STAT-5 mRNA expression (P < 0.05). Glucose concentration and cellular metabolic activity were not different between controls and insulin-treated cells. Based on our results, we suggest that primary hepatocyte cultures can be used to study the effect of LPS on the inflammatory cascade. Insulin decreases hepatic cytokine expression, which is associated with decreased STAT-5 expression.



This study was supported by the American Surgical Association Foundation, Shriners Hospitals for Children 8660, and Deutsche Forschungsgemeinschaft DFG (Je 233/6-1).


  1. 1.
    Carter EA, Burks D, Fischman AJ, White M, Tompkins RG. (2004) Insulin resistance in thermally-injured rats is associated with post-receptor alterations in skeletal muscle, liver and adipose tissue. Int. J. Mol. Med. 14:653–8.PubMedGoogle Scholar
  2. 2.
    Van den Berghe G. (2003) Insulin therapy for the critically ill patient. Clin. Cornerstone 5:56–63.CrossRefPubMedGoogle Scholar
  3. 3.
    Van den Berghe G, et al. (2003) Outcome benefit of intensive insulin therapy in the critically ill: insulin dose versus glycemic control. Crit. Care. Med. 31:359–66.CrossRefPubMedGoogle Scholar
  4. 4.
    Koenig W. (2002) Insulin resistance, heart disease and inflammation. Identifying the ‘at-risk’ patient: the earlier the better? The role of inflammatory markers. Int. J. Clin. Pract. Suppl. Oct:23–30.Google Scholar
  5. 5.
    van den Berghe G, Wouters P, Weekers F, et al. (2001) Intensive insulin therapy in critically ill patients. N. Engl. J. Med. 345:1359–67.CrossRefGoogle Scholar
  6. 6.
    Van den Berghe G, et al. (2006) Intensive insulin therapy in the medical ICU. N. Engl. J. Med. 354:449–61.CrossRefPubMedGoogle Scholar
  7. 7.
    Ellger B, et al. (2006) Survival benefits of intensive insulin therapy in critical illness: impact of maintaining normoglycemia versus glycemia-independent actions of insulin. Diabetes 55:1096–105.CrossRefPubMedGoogle Scholar
  8. 8.
    Cosentino F, et al. (2003) High glucose causes up-regulation of cyclooxygenase-2 and alters prostanoid profile in human endothelial cells: role of protein kinase C and reactive oxygen species. Circulation 107:1017–23.CrossRefPubMedGoogle Scholar
  9. 9.
    Dandona P, et al. (2001) Insulin inhibits intranuclear nuclear factor kappaB and stimulates IkappaB in mononuclear cells in obese subjects: evidence for an anti-inflammatory effect? J. Clin. Endocrinol. Metab. 86:3257–65.PubMedGoogle Scholar
  10. 10.
    Guha M, Bai W, Nadler JL, Natarajan R. (2000) Molecular mechanisms of tumor necrosis factor alpha gene expression in monocytic cells via hyperglycemia-induced oxidant stress-dependent and -independent pathways. J. Biol. Chem. 275:17728–39.CrossRefPubMedGoogle Scholar
  11. 11.
    Yano M, et al. (2004) Short-term exposure of high glucose concentration induces generation of reactive oxygen species in endothelial cells: implication for the oxidative stress associated with postprandial hyperglycemia. Redox. Rep. 9:111–6.CrossRefPubMedGoogle Scholar
  12. 12.
    Thasler WE, Weiss TS, Schillhorn K, Stoll PT, Irrgang B, Jauch KW. (2003) Charitable State-Controlled Foundation Human Tissue and Cell Research: Ethic and legal aspects in the supply of surgically removed human tissue for research in the academic and commercial sector in Germany. Cell Tissue. Bank 4:49–56.CrossRefPubMedGoogle Scholar
  13. 13.
    Thasler WE, et al. (2006) Repression of cytochrome P450 activity in human hepatocytes in vitro by a novel hepatotrophic factor, augmenter of liver regeneration. J. Pharmacol. Exp. Ther. 316:822–9.CrossRefPubMedGoogle Scholar
  14. 14.
    Weiss TS, Pahernik S, Scheruebl I, Jauch KW, Thasler WE. (2003) Cellular damage to human hepatocytes through repeated application of 5-aminolevulinic acid. J. Hepatol. 38:476–82.CrossRefPubMedGoogle Scholar
  15. 15.
    Hoebe KH, Witkamp RF, Fink-Gremmels J, Van Miert AS, Monshouwer M. (2001) Direct cell-to-cell contact between Kupffer cells and hepatocytes augments endotoxin-induced hepatic injury. Am. J. Physiol. Gastrointest. Liver. Physiol. 280:G720–8.CrossRefPubMedGoogle Scholar
  16. 16.
    Zinchenko YS, Schrum LW, Clemens M, Coger RN. (2006) Hepatocyte and kupffer cells co-cultured on micropatterned surfaces to optimize hepatocyte function. Tissue. Eng. 12:751–61.CrossRefPubMedGoogle Scholar
  17. 17.
    Aljada A, Ghanim H, Mohanty P, Kapur N, Dandona P. (2002) Insulin inhibits the pro-inflammatory transcription factor early growth response gene-1 (Egr)-1 expression in mononuclear cells (MNC) and reduces plasma tissue factor (TF) and plasminogen activator inhibitor-1 (PAI-1) concentrations. J. Clin. Endocrinol. Metab. 87:1419–22.CrossRefPubMedGoogle Scholar
  18. 18.
    Aljada A, Ghanim H, Saadeh R, Dandona P. (2001) Insulin inhibits NFkappaB and MCP-1 expression in human aortic endothelial cells. J. Clin. Endocrinol. Metab. 86:450–3.PubMedGoogle Scholar
  19. 19.
    Dandona P, Aljada A, Dhindsa S, Garg R. (2003) Insulin as an anti-inflammatory and antiatherosclerotic hormone. Clin. Cornerstone Suppl 4:S13–20.CrossRefGoogle Scholar
  20. 20.
    Jeschke MG, Einspanier R, Klein D, Jauch KW. (2002) Insulin attenuates the systemic inflammatory response to thermal trauma. Mol. Med. 8:443–50.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Jeschke MG, Klein D, Bolder U, Einspanier R. (2004) Insulin attenuates the systemic inflammatory response in endotoxemic rats. Endocrinology 145:4084–93.CrossRefPubMedGoogle Scholar
  22. 22.
    Jeschke MG, Klein D, Herndon DN. (2004) Insulin treatment improves the systemic inflammatory reaction to severe trauma. Ann. Surg. 239:553–60.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Jeschke MG, et al. (2005) Insulin prevents liver damage and preserves liver function in lipopolysaccharide-induced endotoxemic rats. J. Hepatol. 42:870–9.CrossRefPubMedGoogle Scholar
  24. 24.
    Das UN. (2003) Insulin in sepsis and septic shock. J. Assoc. Physicians. India 51:695–700.PubMedGoogle Scholar
  25. 25.
    Das UN. (2002) Insulin and the critically ill. Crit. Care 6:262–3.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Das UN. (2002) Insulin and inflammation: further evidence and discussion. Nutrition 18:526–7.CrossRefPubMedGoogle Scholar
  27. 27.
    Esposito K, et al. (2002) Inflammatory cytokine concentrations are acutely increased by hyperglycemia in humans: role of oxidative stress. Circulation 106:2067–72.CrossRefPubMedGoogle Scholar
  28. 28.
    Gore DC, Chinkes D, Heggers J, Herndon DN, Wolf SE, Desai M. (2001) Association of hyperglycemia with increased mortality after severe burn injury. J. Trauma 51:540–4.PubMedGoogle Scholar
  29. 29.
    Gore DC, Chinkes DL, Hart DW, Wolf SE, Herndon DN, Sanford AP. (2002) Hyperglycemia exacerbates muscle protein catabolism in burn-injured patients. Crit. Care. Med. 30:2438–42.CrossRefPubMedGoogle Scholar
  30. 30.
    Ling PR, Mueller C, Smith RJ, Bistrian BR. (2003) Hyperglycemia induced by glucose infusion causes hepatic oxidative stress and systemic inflammation, but not STAT3 or MAP kinase activation in liver in rats. Metabolism 52:868–74.CrossRefPubMedGoogle Scholar
  31. 31.
    Jeschke MG, Mlcak RP, Finnerty CC, Herndon DN. (2007) Changes in liver function and size after a severe thermal injury. Shock 28:172–7.CrossRefPubMedGoogle Scholar
  32. 32.
    Finnerty CC, et al. (2006) Cytokine expression profile over time in severely burned pediatric patients. Shock 26:13–9.CrossRefPubMedGoogle Scholar
  33. 33.
    Rivers E, et al. (2001) Early goal-directed therapy in the treatment of severe sepsis and septic shock. N. Engl. J. Med. 345:1368–77.CrossRefGoogle Scholar
  34. 34.
    Rennie MJ. (1985) Muscle protein turnover and the wasting due to injury and disease. Br. Med. Bull. 41:257–64.CrossRefPubMedGoogle Scholar
  35. 35.
    Tracey KJ, et al. (1987) Anti-cachectin/TNF monoclonal antibodies prevent septic shock during lethal bacteraemia. Nature 330:662–4.CrossRefPubMedGoogle Scholar
  36. 36.
    Takala J, et al. (1999) Increased mortality associated with growth hormone treatment in critically ill adults. N. Engl. J. Med. 341:785–92.CrossRefPubMedGoogle Scholar
  37. 37.
    Tracey KJ, et al. (1987) Cachectin/tumor necrosis factor induces lethal shock and stress hormone responses in the dog. Surg. Gynecol. Obstet. 164:415–22.PubMedGoogle Scholar
  38. 38.
    Wang H, et al. (1999) HMG-1 as a late mediator of endotoxin lethality in mice. Science 285:248–51.CrossRefPubMedGoogle Scholar
  39. 39.
    Wang H, et al. (1999) Proinflammatory cytokines (tumor necrosis factor and interleukin 1) stimulate release of high mobility group protein-1 by pituicytes. Surgery 126:389–92.CrossRefPubMedGoogle Scholar
  40. 40.
    Moshage H. (1997) Cytokines and the hepatic acute phase response. J. Pathol. 181:257–66.CrossRefPubMedGoogle Scholar

Copyright information

© Feinstein Institute for Medical Research 2008

Authors and Affiliations

  • Marc G. Jeschke
    • 1
    • 2
  • Dagmar Klein
    • 3
  • Wolfgang E. Thasler
    • 4
  • Ulrich Bolder
    • 3
  • Hans-Jürgen Schlitt
    • 3
  • Karl-Walter Jauch
    • 4
  • Thomas S. Weiss
    • 3
  1. 1.Shriners Hospitals for ChildrenGalveston Burns UnitGalvestonUSA
  2. 2.Department of SurgeryUniversity Texas Medical BranchGalvestonUSA
  3. 3.Department of SurgeryUniversity of Regensburg HospitalRegensburgGermany
  4. 4.Department of SurgeryLM University Munich GrosshadernMunichGermany

Personalised recommendations