Table 4.
Nitric oxide synthase, HSP/heme oxygenase-1, matrix metalloproteinase knockout models of liver ischemia reperfusion injury
| Ref. | Knockout model | IR protocol | Outcome measure | Agent | Adaptive responses | Injurious responses |
| Hamada et al[26] | iNOS (-/-); MMP-9 (-/-) | 70% I 90 min/R 3, 6, 24 h | Histology; serum ALT, NO2-/NO3-; myeloperoxidase activity (MPO); immunohistochemistry; PCR, Western blotting; MMP-9 activity assay; MMP-9 protein levels; neutrophil (PMN) migration assay; TUNEL and caspase-3 activity | ONO-1714 (iNOS inhibitor); NO donor (DETA NONOate) | Increased macrophage iNOS producing NO increases PMN MMP-9 and PMN transmigration over fibronectin | |
| Hamada et al[42] | MMP-9 (-/-) | 70% I 90 min/R 6, 24 h | Histology; serum GPT and GOT; MPO; IH; PCR | Anti MMP-9 iv; MMP-2/9 inhibitor; anti MMP-2 (all to WT only) | MMP-9 (not MMP-2) increase TNF-α, IFNg, IL2, IL6 and increase PMN and CD4+ T cell recruitment leading to increased liver necrosis | |
| Kuboki et al[73] | HSP70 (-/-) | 70% I 90 min/R 1, 8 h | Histology; serum AST; TNF-α; IL6; MIP-2; MPO; WB; EMSA (NFκβ) | Sodium arsenite iv to induce HSP70; recombinant HSP70 | No involvement of HSP70 in IRI; NFκβ activity associated with IRI | |
| Theruvath et al[59] | eNOS (-/-) | Donor (WT/KO) to WT recipient; organ stored 18 h, 4 °C, UWS | Histology; serum ALT; IVM; TUNEL; IH (macrophage infiltration) | eNOS activation reduces necrosis and apoptosis, with associated inhibition of macrophage infiltration, increased sinusoidal diameter and blood flow | ||
| Tsuchiashi et al | HO-1 (+/-); HO-1 (-/-) | 70% I 90 min/R 6 h | Histology; serum GOT; MPO; quantitative real time RT-PCR; WB; TUNEL | CoPP (induces HO-1) 24 h preop | HO-1 upregulated which inhibits expression of cytokines TNF-α and IFNγ | TNF-α and IFNγ expression increased overall in IRI associated with increased apoptosis and necrosis |
| Hines et al[23] | eNOS (-/-); iNOS (-/-) | 70% I 45 min/R 1, 3 h | Serum ALT; histology; PCR | Increased eNOS expression in IRI inhibits TNF-α and IL12 expression; iNOS activates eNOS in this model | No PMN infiltration at 3 h reperfusion | |
| Lee et al[58] | eNOS (-/-); iNOS (-/-) | 70% I 1 h/R 1, 3, 6 h | Serum ALT and AST; perfusion studies; PCR | eNOS activated during IRI is protective | Increased iNOS mRNA expression from 3 h reperfusion onwards regulates reperfusion and is associated with worse IRI | |
| Hines et al[60] | iNOS (-/-) | 70% I 45 min/R 1, 3, 6 h | Serum ALT; histology; MPO | L-NIL (iNOS inhibitor) | Reduced IRI in iNOS (-/-), but no iNOS mRNA or L-NIL effect in WT; may be genetic compensation effect in KO | |
| Kawachi et al[24] | eNOS (-/-); iNOS (-/-) | 70% I 45 min/R 5 h | Serum ALT; histology; MPO | eNOS is activated in IRI and is protective | There is no PMN infiltration up to 5 h reperfusion and iNOS is not activated in IRI in this model |
IH: Immunohistochemistry; WB: Western blotting; MPO: Myeloperoxidase assay; RT-PCR: Reverse transcriptase polymerase chain reaction; ELISA: Enzyme labelled immunosorbent assay; EMSA: Electrophoretic mobility shift assay; AST: Aspartate transaminase; ALT: Alanine transaminase; GOT: Glutamic oxaloacetic 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; MMP: Matrix metalloproteinase; HSP: Heat shock protein; eNOS: Endothelial nitric oxide synthase; iNOS: Inducible nitric oxide synthase; HO-1: Heme oxygenase (subtype 1).