This paper is only available as a PDF. To read, Please Download here.
Abstract
Background. Sustained intestinal ischemic injury often leads to shock and multiorgan failure,
mediated in part by a cytokine cascade. Animal models have also identified a central
role of Kupffer cells in amplification of cytokines following intestinal ischemia.
To better understand this gut–liver axis, we developed an in vitro model.
Materials and methods. Kupffer cells were isolated from rat livers by arabinogalactan gradient ultracentrifugation
and adherence purification. Cells were grown in RPMI medium in 5% CO2. Rat intestinal epithelial cells, IEC-6, were cultured under normoxic or anoxic (90%
N2, 10% CO2) conditions for 2, 12, and 24 h. Kupffer cells were then grown in the conditioned
medium of the IEC-6 cultures. After 24 h, the medium was replaced with fresh medium.
This final Kupffer cell supernatant was tested for tumor necrosis factor α and interleukin-6
production by ELISA. Trypan blue exclusion was performed to assess cell viability.
Results. Intestinal and Kupffer cells remained viable during the experimental time. Production
of both tumor necrosis factor α and interleukin-6 by Kupffer cells increased with
increasing ischemia time of the intestinal cells.
Conclusions. Consistent with animal studies of intestinal ischemia, this study found an increase
in cytokine production by Kupffer cells following hypoxia of intestinal cells. This
in vitro model offers a new tool to study the expression of cytokines, proteins, and messengers
involved in the cascade of events that follow intestinal ischemia.
Keywords
To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to Journal of Surgical ResearchAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
REFERENCES
- The role of intestinal barrier failure and bacterial translocation in the development of systemic infection and multiple organ failure.Arch. Surg. 1990; 125: 403
- History of mesenteric ischemia: The evolution of a diagnosis and management.Surg. Clin. North Am. 1997; 77: 275
- Intestinal ischemic disorders.Dig. Dis. 1995; 13: 3
- Seatbelt sign following blunt trauma is associated with increased incidence of abdominal injury.Am. Surg. 1997; 63: 885
- Factors related to outcome in blunt intestinal injuries requiring operation.Am. Surg. 1997; 63: 889
- Intestinal injury mechanisms after blunt abdominal impact.Ann. R. Coll. Surg. Engl. 1997; 79: 115
- Gastrointestinal complications after aortic surgery.J. Vasc. Surg. 1998; 28: 404
- Acute mesenteric ischemia after open heart surgery.Angiology. 1998; 49: 267
- Perioperative predictors of colonic ischemia after ruptured abdominal aortic aneurysm.J. Vasc. Surg. 1999; 29: 40
- Nonocclusive intestinal ischemia: Improved outcome with early diagnosis and therapy.Am. Surg. 1997; 63: 334
- Improved outcome by identification of high-risk nonocclusive mesenteric ischemia, aggressive reexploration, and delayed anastomosis.Am. J. Surg. 1995; 170: 577
- “Sepsis/SIRS”, physiologic classification, severity stratification, relation to cytokine elaboration and outcome prediction in posttrauma critical illness.J. Trauma. 1996; 41: 581
- MOF/MODS, SIRS: An update.Shock. 1996; 6: S1
- Systemic inflammatory response syndrome (SIRS), multiple organ dysfunction syndrome (MODS), multiple organ failure (MOF): Are we winning the battle.Shock. 1998; 10: 79
- Gut ischemia and mesenteric synthesis of inflammatory cytokines after hemorrhagic or endotoxic shock.Am. J. Physiol. 1997; 273: G314
- Cytokine gene expression in splenic macrophages and Kupffer cells following haemorrhage.Cytokine. 1995; 7: 8
- Hypoxemia in the absence of blood loss or significant hypotension causes inflammatory cytokine release.Am. J. Physiol. 1995; 269: R160
- Hypoxia induces a human macrophage cell line to release tumor necrosis factor-alpha and its soluble receptors in vitro.J. Surg. Res. 1993; 54: 281
- Cellular and Molecular Immunology. W.B. Saunders, Philadelphia1991
- Cytokine patterns in patients after major vascular surgery, hemorrhagic shock, and severe blunt trauma: Relation with subsequent adult respiratory distress syndrome and multiple organ failure.Ann. Surg. 1993; 218: 769
- Tumor necrosis factor in sepsis: Mediator of multiple organ failure or essential part of host defense.Shock. 1995; 3: 1
- Efficacy and safety of monoclonal antibody to human tumor necrosis factor alpha in patients with sepsis syndrome: A randomized, controlled, double-blind, multicenter clinical trial. TNF-alpha MAb Sepsis Study Group.JAMA. 1995; 273: 934
- Passive immunization against cachectin/tumor necrosis factor protects mice from lethal effect of endotoxin.Science. 1985; 229: 869
- Evolving strategies in the treatment of sepsis and systemic inflammatory response syndrome (SIRS).Q. J. Med. 1998; 91: 265
- Neutrophil adherence to isolated adult cardiac myocytes: Induction by cardiac lymph collected during ischemia and reperfusion.J. Clin. Invest. 1992; 89: 602
- Heterogeneity of Kupffer cells and splenic, alveolar, and peritoneal macrophages for the production of TNF, IL-1, and IL-6.Inflammation. 1994; 18: 511
- Role of Kupffer cells in interleukin-6 release following trauma: Hemorrhage and resuscitation.Shock. 1994; 1: 43
- Enhancement of hepatic macrophages in septic rats and their inhibitory effect on hepatocyte function.J. Surg. Res. 1991; 50: 72
- Enhancement of rat hepatic macrophages by treatment with interleukin-2 and streptococcal preparation OK432, with reference to antitumor activity, soluble factor production and Ia expression.Cancer Immunol. Immunother. 1992; 35: 75
Rigberg, D. A, Lane, J. S, Todd, K. E, Centeno, J, Gloor, B, McFadden, D. W, and, Chandler, C. Kupffer cell manipulation influences systemic cytokines in intestinal ischemia/reperfusion, Unpublished data,, 1999.
- Cytokine gene expression by Kupffer cells in experimental alcoholic liver disease.Hepatology. 1995; 22: 1304
Article info
Footnotes
☆Presented at the 23rd Annual Symposium of the Association of Veterans Administration Surgeons, St. Louis, MO, May 2–4, 1999.
☆☆This study was supported by a VA Merit Review Grant (D.W.M.).
Identification
Copyright
© 2000 Academic Press. Published by Elsevier Inc. All rights reserved.
