Table 5.
Knockouts of downstream mediators and models of liver ischemia reperfusion injury
| Ref. | Knockout model | IR protocol | Outcome measure | Agent | Adaptive responses | Injurious responses |
| Theruvath et al[14,15] | JNK (-/-) | 70% I 1 h/R 4, 8 h | Histology; serum ALT; IVM (dyes to probe mitochondrial function and cell death); survival 14 d | JNK2 activation leads to mitochondrial depolarization and increased necrosis only | ||
| Theruvath et al[14,15] | JNK (-/-) | WT or KO donor; 30 h, 4 °C, UWS preservation; WT recipient | Histology 8 h posttransplant; serum ALT, TUNEl and caspase-3 assay; IVM; IH; lipid peroxidation | JNK2 activation leads to caspase-3 activation, mitochondrial depolarisation and release of cytochrome c, lipid peroxidation which all translate into reduced survival posttransplant | ||
| Beraza et al[72] | Conditional hepatocyte specific NEMO knockout | 70% I 1 h (caudate lobe resected)/R 3, 6 h | TUNEL and caspase-3 assay; WB; IH; Southern blotting; EMSA (NFκβ) | NFκβ activity reduces necrosis and apoptosis, inhibits TNF-α, JNK and iNOS | ||
| Okaya et al[35] | PPARα (-/-) | 70%I 90 min/R 4, 8 h | Serum ALT; TNF-α; MIP-2; MPO; liver NO2/NO3; WB; EMSA (AP-1, NFκβ) | WY14643 iv (PPARα agonist) | PPARα protective in IRI | PPARα independent release of TNF-α and MIP-2 and increased NO2/NO3 associated with IRI |
| Shen et al[78] | STAT4 (-/-); STAT6 (-/-); nu/nu | 70%I 90 min/R 6 h | Serum ALT; histology; MPO; WB; PCR | Adoptive transfer of CD4+ T cells from WT or other KO to nu/nu; SnPP ip | HO-1 expressed at very low levels after 6 h in this model, but protective in IRI | CD4+T cell activation involving T cell STAT4 activation, but not STAT6 associated with increased IRI |
| Khandoga et al[61] | PARP (-/-) | Left lobe I 90 min/R 30 min | Serum ALT; IVM; IH; PCR | PARP activation in IRI upregulates E-selectin, ICAM1 and VCAM1, associated with increased platelet and leucocyte endothelial interaction and reduced sinusoidal perfusion | ||
| Kato et al[82] | P50 NFκβ (-/-) | 70% I 90 min/R 1, 8 h | Serum ALT; histology; MPO WB; EMSA (p50 and p65 subunits of NFκβ) | No effect of p50 subunit deletion, but increased p50/p65 heterodimer in WT and some p65 in KO, so there may be some functional redundancy of NFκβ subunits | ||
| Kato et al[83] | STAT4 (-/-) | 70% I 90 min/R 30 min, 1, 2, 4, 8 h | Serum ALT; histology; MPO; WB | Anti IL12 Ab | IL12 expression associated with IRI. STAT4 not activated in IRI in this model | |
| Kato et al[70] | STAT6 (-/-) | 70% I 90 min/R 1, 4, 8 h | Serum ALT, TNF-α; MPO; PCR; EMSA (NFκβ) | IL4 or IL13 iv | STAT6 is not activated in IRI in this model, although STAT6 activation by iv IL4 or IL13 is protective. IRI is associated with increased NFκβ DNA binding |
KO: Transgenic knockout; WT: Wild type (normal animals); IH: Immunohistochemistry; WB: Western blotting; MPO: Myeloperoxidase assay; PCR: Polymerase chain reaction; EMSA: Electrophoretic mobility shift assay; ALT: Alanine transaminase; I: Ischemia; R: Reperfusion; IR: Ischemia reperfusion; IRI: Ischemia reperfusion injury; IVM: Intravital microscopy; IFN: Interferon; Ab: Antibody; TNF: Tumour necrosis factor; TNFR1: Tumour necrosis factor receptor (subtype 1); TUNEL: Terminal deoxynucleotidyl transferase dUTP nick end labeling (assay for cell death); IL: Interleukin; NF: Nuclear factor; STAT: Signal Transducer and Activator of Transcription; JNK: A survival kinase; PARP: Poly (ADP-ribose) polymerase.