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Cited by 821Organ Transplantation Techniques and OutcomesLiver physiology and pathologyCardiac Ischemia and Reperfusion

Role of tumor necrosis factor-alpha in the pathophysiologic alterations after hepatic ischemia/reperfusion injury in the rat.

Open access
Colletti, L M, Remick, D G, Burtch, G D, Kunkel, S L, Strieter, R M, Campbell, D A·1990·Journal of Clinical Investigation
DOI 10.1172/jci114656PMC PMC296661
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Abstract

Cytokines are recognized as critical early mediators of organ injury. We attempted to determine whether or not severe hepatic ischemia/reperfusion injury results in tumor necrosis factor-alpha (TNF-alpha) release with subsequent local and systemic tissue injury. After 90 min of lobar hepatic ischemia, TNF was measurable during the reperfusion period in the plasma of all 14 experimental animals, with levels peaking between 9 and 352 pg/ml. Endotoxin was undetectable in the plasma of these animals. Pulmonary injury, as evidenced by a neutrophilic infiltrate, edema and intra-alveolar hemorrhage developed after hepatic reperfusion. The neutrophilic infiltrate was quantitated using a myeloperoxidase (MPO) assay; this demonstrated a significant increase in MPO after only 1 h of reperfusion. Anti-TNF antiserum pretreatment significantly reduced the pulmonary MPO after hepatic reperfusion. After a 12-h reperfusion period, there was histologic evidence of intra-alveolar hemorrhage and pulmonary edema. Morphometric assessment showed that pretreatment with anti-TNF antiserum was able to completely inhibit the development of pulmonary edema. Liver injury was quantitated by measuring serum glutamic pyruvic transaminase which showed peaks at 3 and 24 h. Anti-TNF antiserum pretreatment was able to significantly reduce both of these peak elevations. These data show that hepatic ischemia/reperfusion results in TNF production, and that this TNF is intimately associated with pulmonary and hepatic injury.

AI snapshot

Written from the paper’s full text.

TL;DR

In a rat model of 90-minute lobar hepatic ischemia/reperfusion, TNF became measurable in plasma in all 14 experimental animals and was associated with subsequent pulmonary and hepatic injury. Pretreatment with anti-TNF antiserum reduced pulmonary neutrophil sequestration, abolished pulmonary edema, and lowered serum SGPT elevations after reperfusion.

Key findings

  • After hepatic ischemia/reperfusion, TNF was measurable in plasma of all 14 experimental animals, with a mean peak level of 69.4±30.3 pg/ml and a range of 9–352 pg/ml; TNF was undetectable in sham-operated controls, and endotoxin was undetectable (<5 pg/ml).
  • Pulmonary neutrophil sequestration, measured by MPO, peaked after 1 h of reperfusion at 0.653±0.089 U versus 0.213±0.034 U in sham animals (P<0.005). Anti-TNF antiserum reduced MPO to 0.458±0.057 U compared with 0.701±0.040 U in control-serum animals (P<0.005), though it did not return values to sham baseline.
  • After 12 h of reperfusion, control-serum animals had 20±4 flooded alveoli/high-power field, whereas anti-TNF antiserum-treated animals had 0±0 (P<0.05 by Wilcoxon rank-sum test).
  • Serum SGPT peaked at 3 and 24 h after reperfusion; anti-TNF antiserum significantly reduced both peak elevations (P<0.05 at 3 h and P<0.005 at 24 h versus untreated and control-serum groups). Twenty-four-hour survival was 67% in untreated animals, 92% with anti-TNF antiserum, and 60% with control serum, but the differences were not statistically significant.

Methods

  • Adult male pathogen-free Sprague-Dawley rats (200–300 g) underwent 90 min of lobar hepatic ischemia using a Heifitz clip, with sham-operated controls treated identically except for vascular occlusion.
  • Plasma TNF activity was measured with a WEHI 164 subclone 13 bioassay; positive samples were confirmed by neutralization with polyclonal rabbit anti-murine TNF antiserum. Endotoxin was measured by a perchloric acid-modified chromogenic limulus assay.
  • Organ injury was assessed by pulmonary MPO assay, blinded histologic morphometry of edema-filled alveoli, and serum SGPT. Anti-TNF effects were tested by pretreating rats with polyclonal rabbit anti-murine TNF antiserum versus control rabbit serum. Statistical comparisons used Student Newman-Keuls or Wilcoxon rank-sum tests when data were not normally distributed.

Limitations

  • The paper states that 24-h survival differences were not statistically significant, most likely because of relatively small group sizes.
  • The paper states that further studies are needed to document the degree to which the observed pulmonary pathology is neutrophil dependent.
  • The paper states that the mechanism of TNF induction after hepatic ischemia/reperfusion is not completely clear; the text infers Kupffer cell production/leakage but does not directly establish the cellular source.
  • Evident from the text: anti-TNF antiserum did not prevent the transient hypotension after revascularization, so the protective effects observed were not attributable to hemodynamic stabilization.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1172/jci114656

Cites

42 of 42 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 16 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    After 90 min of lobar hepatic ischemia and reperfusion, TNF was measurable in the plasma of all 14 experimental rats.

    “After 90 min of lobar hepatic ischemia, TNF was measurable during the reperfusion period in the plasma of all 14 experimental animals, with levels peaking between 9 and 352 pg/ml.”
    • Also stated in: § Results · plain
      “TNF was measurable in the plasma of all 14 experimental animals”
    Quote in the paperSubstantial claim
  • Findingown data

    Endotoxin was undetectable (<5 pg/ml) in the plasma of rats after hepatic ischemia/reperfusion.

    “Endotoxin was undetectable in the plasma of these animals.”
    • Also stated in: § Results · plain
      “In addition, endotoxin was unde- tectable (< 5 pg/ml).”
    Quote in the paperSubstantial claim
  • Findingown data

    TNF was undetectable at any time point in the plasma of three sham-operated control rats.

    “TNF was un- detectable at any time point in the three sham-operated con- trol animals”
    Quote in the paperSubstantial claim
  • Findingown data

    Pulmonary MPO was significantly increased after 1 h of reperfusion compared with sham-operated controls (0.653±0.089 U vs 0.213±0.034 U, P<0.005).

    “The neutrophilic infiltrate was quantitated using a myeloperoxidase (MPO) assay; this demonstrated a significant increase in MPO after only 1 h of reperfusion.”
    • Also stated in: § Results · plain
      “1 h postrevascularization, mean pulmonary MPO was 0.653±0.089U as compared with 0.213±0.034U for sham-operated animals (P < 0.005; Fig. 3.).”
    Quote in the paperSubstantial claim
  • Findingown data

    Pulmonary injury, evidenced by neutrophilic infiltrate, edema, and intra-alveolar hemorrhage, developed after hepatic reperfusion.

    “Pulmonary injury, as evidenced by a neutrophilic infiltrate, edema and intra-alveolar hemorrhage developed after hepatic reperfusion.”
    Quote in the paperSubstantial claim
  • May combine two claims. 17
  • The quote only partly states this claim. 3
  • Classified as probably not a substantial claim. 7

  • May combine two claims.

    Findingown data

    The mean peak plasma TNF level after hepatic ischemia/reperfusion was 69.4±30.3 pg/ml, with a range of 9-352 pg/ml.

    “The mean peak TNF level was 69.4±30.3pg/ml with a range of 9-352 pg/ml.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Anti-TNF treatment blunted the increase in pulmonary MPO but did not reduce MPO to sham-operated control levels.

    “Thus, anti-TNF treatment blunted the increase in pulmonary MPO but did not reduce the MPO level to that observed in sham-operated controls.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Pulmonary MPO levels were maximal after 1 h of reperfusion and gradually decreased over the next 11 h.

    “MPO levels were maximal after 1 h of reperfusion and gradually decreased over the next 11 h.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    24-h survival in control serum-treated animals was similar to the untreated experimental group, with 6 of 10 animals surviving for 24 h.

    “24 h survival in animals treated with control serum was similar to that seen in the untreated experimental group with 6 of 10 animals surviving for 24 h.”
    Quote in the paperNot a substantial claim

  • The quote only partly states this claim.

    Findingown data

    Anti-TNF antiserum almost completely abrogated the microvascular leak and hemorrhagic edema in the lung.

    “the anti-TNF-treated rats illustrate essentially normal histology.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Anti-TNF antiserum pretreatment significantly reduced both peak SGPT elevations at 3 and 24 h after reperfusion.

    “Anti-TNF antiserum pretreatment was able to significantly reduce both of these peak elevations.”
    • Also stated in: § Results · plain
      “SGPT levels were significantly decreased both 3 and 24 h postrevascularization in the animals treated with anti-TNF antiserum as compared to untreated experimental animals and animals treated with control serum (P < 0.05 after 3 h of reperfusion and”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Statistical significance for 24-h survival was not achieved between any of the groups, most likely due to the relatively small numbers in the groups.

    “Statistical significance was not achieved between any of the groups, most likely due to the relatively small numbers in the groups.”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    Anti-TNF antiserum pretreatment significantly decreases SGPT after 3 h and 24 h of hepatic reperfusion compared with untreated and control serum-treated animals.

    “After 3 h of hepatic reperfusion, there was a significant decrease in SGPT in animals pretreated with anti-TNF antiserum as compared with both the untreated ex- perimental animals and the control serum-treated animals.”
    • Also stated in: § Discussion · hedged
      “Animals treated with anti-TNF antiserum showed a significant reduction in a hepatocyte-specific enzyme (SGPT) after hepatic ischemia/reperfusion in comparison to controls treated with control serum”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    Anti-TNF antiserum pretreatment significantly reduced pulmonary MPO after hepatic reperfusion compared with control serum-treated animals (0.458±0.057 U vs 0.701±0.040 U, P<0.005).

    “Anti-TNF antiserum pretreatment significantly reduced the pulmonary MPO after hepatic reperfusion.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Operated rats after hepatic ischemia/reperfusion show intra-alveolar hemorrhage and some edema fluid in the lung.

    “the operated rats show intra-alveolar hemorrhage and some edema fluid.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim. May combine two claims.

    Premisecitation

    Impaired hepatic function greatly influences severity and impedes resolution of ARDS.

    “Recent clinical data have shown that impaired hepatic function greatly influences severity and impedes reso- lution of ARDS (4).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [4]

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    The pulmonary circulation is the first vascular bed into which cytokines from injured liver are delivered, so pulmonary parenchyma is likely to manifest toxic effects of these cytokines.

    “Since the pulmonary circulation represents the first vascu- lar bed into which these cytokines are delivered, the pulmonary parenchyma would be likely to manifest the toxic effects of these cytokines.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The pulmonary circulation is the first vascular bed into which cytokines from injured liver are delivered.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    The lower limit of endotoxin detection in this assay is 5 pg/ml.

    “The lower limit of endotoxin detection in this assay is 5 pg/ml.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    TNF release did not appear directly related to the hypotensive effect produced by the ischemia/reperfusion process, because anti-TNF antiserum treatment blocked the observed pathology but did not alter the hypotension following clamp release.

    “The TNF release also did not appear directly related to the hypotensive effect produced by the I/R process, since anti-TNF antiserum treatment blocked the observed pathology, but did not alter the hypotension following clamp release.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    Primary nonfunction of the graft after liver transplantation is associated with rapid development of pulmonary insufficiency.

    “primary nonfunction of the graft after liver transplantation is also associated with the rapid development ofpulmonary insufficiency (5).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [5]

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    The pulmonary pathology after hepatic ischemia/reperfusion appears to be a consequence of TNF, because pretreatment with neutralizing anti-TNF antiserum blunted the neutrophilic infiltrate and completely abolished both the diffuse edema and patchy hemorrhage.

    “The pulmonary pathology appears to be a consequence of TNF, since pretreatment with neutralizing anti-TNF anti- serum blunted the neutrophilic infiltrate (as measured by MPO) and completely abolished both the diffuse edema and patchy hemorrhage.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    Slides were examined in a blinded manner.

    “Slides were examined in a blinded manner.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Previous in vitro studies have documented the ability of Kupffer cells to produce TNF.

    “Previous in vitro studies have docu- mented the ability of Kupffer cells to produce TNF (28).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [28]

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    The anti-TNF antiserum neutralizes both natural and recombinant TNF, cross-reacts with rat TNF, and has no cross-reactivity with interleukin 1.

    “This antiserum neutralizes both natural and recombinant TNF, cross-reacts with rat TNF and has no cross-reactivity with interleukin 1.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites 1 1, 12

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Previous studies have documented a deleterious effect of TNF on the hepatocyte, where infused rTNF results in focal hepatic necrosis, as does LPS infusion, and the latter finding could be blocked with anti-TNF antiserum.

    “Previous studies have documented a deleterious effect of TNF on the hepatocyte where infused rTNF results in focal hepatic necrosis, as does LPS infusion, and the latter finding could be blocked with anti-TNF antiserum (13).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [13]

      Not checked in the free run.

  • May combine two claims.

    Study design and conductnone given

    The model was specifically chosen to avoid vascular congestion, involving only lobar vascular occlusion with unimpeded portal flow to the right lobe.

    “However, our model was specifically chosen to avoid such vascular con- gestion, involving only lobar vascular occlusion with unim- peded portal flow to the right lobe.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisereasoning

    The three caudal lobes retain an intact portal inflow and venous outflow, preventing intestinal venous congestion and possible leakage of bacteria or bacterial products into the circulation.

    “The three caudal lobes retain an intact portal inflow and venous out- flow, preventing intestinal venous congestion and possible leakage of bacteria or bacterial products into the circulation.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationown data

    The data confirm the earlier study that used an intravenous radioactive tracer to document protein leakage into the lungs.

    “These data confirm our earlier study which used an intravenous ra- dioactive tracer to document protein leakage into the lungs (9).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [9]

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    The rat model of lobar, rather than total, hepatic ischemia produces a severe ischemic insult without inducing mesenteric venous hypertension.

    “By utilizing a rat model of lobar, rather than total, hepatic ischemia, a severe ischemic insult was produced without in- ducing mesenteric venous hypertension (8).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [8]

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    TNF has both local deleterious effects on the hepatocyte and extensive damaging effects on pulmonary integrity.

    “and that TNF has both local deleterious effects on the hepatocyte and extensive damaging effects on pulmonary integrity.”
    Quote in the paperSubstantial claim
25 of 25

  • The paper relies on this without support.

    TNF is produced primarily by cells of the monocyte/macrophage lineage.

    “TNF is produced primarily by cells of the monocyte/macrophage lineage”

  • The paper relies on this without support.

    The liver Kupffer cell mass is the largest fixed macrophage population in the body.

    “the liver Kupffer cell mass is the largest fixed macrophage population in the body.”

  • The paper relies on this without support.

    The pulmonary circulation is the first vascular bed into which cytokines from injured liver are delivered.

    “the pulmonary circulation represents the first vascu- lar bed into which these cytokines are delivered”

  • The paper relies on this without support.

    The anti-TNF antiserum used in these experiments neutralizes TNF in vivo and does not have other relevant biological activities that could account for the observed protection.

    “pretreatment with neutralizing anti-TNF anti- serum blunted the neutrophilic infiltrate”
4 of 4

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Reperfusion of ischemic tissues stimulates cytokine overproduction, including TNF, IL-1, and HMGB1, which contributes to the pathogenesis of resulting tissue damage.

    “Reperfusion of ischemic tissues stimulates cytokine overproduction (e.g., TNF, IL-1, HMGB1), which contributes to the pathogenesis of resulting tissue damage (42)(43)(44).”

    From Physiology and immunology of the cholinergic antiinflammatory pathway (2007) · cites this paper as [43]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Hepatocyte-injury claim cites the Limulus-assay reference

    Minor

    The discussion cites reference 13 for prior findings about TNF, LPS, and hepatic necrosis, but reference 13 describes a method for the Limulus test rather than those findings.

  • Allograft-rejection citation points to a parasitic-infection paper

    Minor

    The text cites reference 24 for TNF in renal allograft rejection, but reference 24 is titled as a study of parasitic infections. The renal allograft rejection paper appears as reference 25.

2 of 2

Cites

Flagged first, then by citations

1 of the 43 references couldn’t be matched to a known paper.

Research works printed but not linked: 2

  1. Glantz, S. A. 1987. Primer of Biostatistics. 2nd edition.
  2. Bevilacqua, M. P., J. S. Pober, G. R. Majeau, W. Fiers, R. S. Cotran, and M. A. Gimbrone. 1986. Recombinant tumor necrosis factor induces procoagulant activity in cultured human vascular endo- thelium. Proc. Natl. Acad. Sci. USA. 83:4533-4537.

Cited by

Sorted by citations

Related

Authors
Colletti, L. M., Remick, D. G., Burtch, G. D., Kunkel, S. L., Strieter, R. M., & Campbell, D. A.
Affiliations
University of Michigan (US)
ORCID
Venue
Journal of Clinical Investigation
Publisher
American Society for Clinical Investigation
Published
1990-06-01
Volume
85
Issue
6
Pages
1936-1943
ISSN
0021-9738 (ISSN-L)
DOI
10.1172/jci114656
PMC
PMC296661
OpenAlex
W2099883737
Semantic Scholar
ef00ab3fc75d2591f58ef3691dd24619b03cf25e
Cited by
817
References
43
Citations by year
2026: 3 · 2025: 2 · 2024: 2 · 2023: 5 · 2022: 10 · 2021: 8 · 2020: 5 · 2019: 6 · 2018: 16 · 2017: 12 · 2016: 6 · 2015: 15 · 2014: 12 · 2013: 20 · 2012: 24
FWCI
40.45
Type
journal article
Language
en
Open access
Locations
  • Journal of Clinical Investigation (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Institutes of Health (5r01hl031237-32)
  • National Heart, Lung, and Blood Institute (HL-35276, HL31237, HL 31963, P01HL031963)
Topics
  • Health Sciences › Medicine › Surgery › Organ Transplantation Techniques and Outcomes
  • Health Sciences › Medicine › Hepatology › Liver physiology and pathology
  • Health Sciences › Medicine › Pathology and Forensic Medicine › Cardiac Ischemia and Reperfusion
Keywords
TNF-α, hepatic ischemia-reperfusion injury, cytokines, pulmonary edema, intra-alveolar hemorrhage, neutrophil infiltration, myeloperoxidase, liver injury, systemic inflammatory response, endotoxin
MeSH terms
Alanine Transaminase, Animals, Hemodynamics, Ischemia, Liver, Liver Diseases, Lung, Peroxidase, Neutrophils, Time Factors, Tumor Necrosis Factor-alpha, Reperfusion Injury, Rats

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Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

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