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Improved myocardial ischemia/reperfusion injury in mice lacking tumor necrosis factor-

Naoya Maekawa, BSc^*, Hisayasu Wada, MD, PhD^*,*, Tsugiyasu Kanda, MD, PhD, Tamikazu Niwa, BSc^*, Yasuhiro Yamada, MD, PhD^*, Kuniaki Saito, PhD^*, Hisayoshi Fujiwara, MD, PhD, Kenji Sekikawa, PhD and Mitsuru Seishima, MD, PhD^*

^* Department of Laboratory Medicine, Gifu University School of Medicine, Gifu, Japan
Second Department of Internal Medicine, Gifu University School of Medicine, Gifu, Japan
Department of Laboratory Medicine, Gunma University School of Medicine, Maebashi, Japan
Department of Immunology, National Institute of Animal Health, Tsukuba, Japan

View larger version (14K): [in a new window]   Figure 1 Arrhythmia observed during myocardial ischemia/reperfusion. (A) Summarized data on the arrhythmia during reperfusion in wild-type (n = 11) and tumor necrosis factor (TNF)- knockout (KO) (n = 9) mice. (B) Summarized data on the arrhythmia during reperfusion in wild-type mice receiving nonimmune IgG (n = 5) and anti-TNF- antibody (n = 5). **p < 0.01.

View larger version (17K): [in a new window]   Figure 2 Hemodynamic parameters measured just before the end of reperfusion. Summarized data on peak left ventricular systolic pressure (LVSP) (A) and maximal positive rate of the first derivative of left ventricular pressure (dP/dtmax) (B) in the sham-operated wild-type mice (solid bars) (n = 5), tumor necrosis factor (TNF)- knockout (KO) mice (light bars) (n = 5), reperfused wild-type (n = 6) and TNF- KO (n = 5) mice. *p < 0.05. I/R = ischemia/reperfusion.

View larger version (56K): [in a new window]   Figure 3 Infarct area (IA) and area at risk (AAR) of the left ventricle (LV) after 30-min ligation and 120-min reperfusion. Representative photographs of midventricular myocardium from wild-type mice(A), tumor necrosis factor (TNF)- knockout (KO) mice (B), wild-type mice receiving nonimmune IgG (C) and wild-type mice receiving anti-TNF- antibody (D). (E) Summarized data on infarct area to area at risk ratio (IA/AAR), IA to left ventricle ratio (IA/LV) and AAR to LV ratio (AAR/LV) in wild-type (n = 11) and TNF- KO (n = 9) mice. (F) Summarized data on IA/AAR, IA/LV and AAR/LV in wild-type mice receiving nonimmune IgG (n = 5) and anti-TNF- antibody (n = 5). **p < 0.01.

View larger version (24K): [in a new window]   Figure 4 Electrophoretic gel mobility shift assay for activation of nuclear factor-B (NF-B). (A) Representative photographs. Lane 1 = reperfused myocardium from wild-type mice; lane 2 = reperfused myocardium from tumor necrosis factor (TNF)- knockout (KO) mice; lane 3 = reperfused myocardium from wild-type mice with competitive oligonucleotides. (B) Summarized data on activation of NF-B in wild-type mice (n = 5) and TNF- KO mice (n = 5). *p < 0.01.

View larger version (31K): [in a new window]   Figure 5 The results of messenger ribonucleic acid expression in wild-type mice (n = 5) and tumor necrosis factor (TNF)- knockout (KO) mice (n = 6) determined by reverse transcription and polymerase chain reaction. (A) TNF- (p = 0.007). (B) Interleukin (IL)-6 (p-values for the presence of TNF-, ischemia and their interaction are 0.118, 0.005 and 0.129, respectively). (C) intercellular adhesion molecule (ICAM)-1 (p = 0.008, 0.002 and 0.026, respectively). (D) macrophage inflammatory protein (MIP)-1 (p = 0.144, 0.037 and 0.139, respectively). (E) MIP-2 (p = 0.016, 0.006 and 0.017, respectively). (F) monocyte chemoattractant protein (MCP)-1 (p = 0.009, 0.004 and 0.019, respectively).

View larger version (138K): [in a new window]   Figure 6 Immunostaining of tumor necrosis factor- in the reperfused myocardium of wild-type mice. Tumor necrosis factor- immunoreactivity was detected in not only myocytes but also infiltrating leukocytes (arrow heads) and endothelial cells (arrows) at x1,000 magnification. Bars = 20 µm.