Dual roles of tumor necrosis factor-alpha receptor-1 in a mouse model of hindlimb ischemia

Vasc Med. 2009 Feb;14(1):37-46. doi: 10.1177/1358863X08098143.

Abstract

Signals in the tumor necrosis factor alpha (TNF-alpha) pathway are upregulated after ischemia, yet its role in peripheral ischemia remains unclear. We investigated the effect of TNF-alpha receptor 1 (TNFR-1) in acute limb ischemia of TNFR-1 knockout (TNFR-1-/-) and wild type (WT, TNFR-1+/+) mice. Laser Doppler scanning showed that although pre-ischemia blood flow levels were similar in these mice, the limb reperfusion after ischemia was significantly higher in TNFR-1-/- mice 1-7 days after injury. Consistently, fewer TUNEL-positive cells, less DNA fragmentation, and a lower ischemic score were detected in the TNFR-1-/- group when compared to WT controls. Western blot analysis revealed less expression of pro-apoptotic markers Bax and cleaved caspase-3 in TNFR-1-/- mice 1 day after ischemia, supporting the hypothesis that the absence of TNFR-1 results in a reduction of apoptosis. The rate of post-ischemia amputation was 50% in WT mice versus 0% in TNFR-1-/- mice. However, immunohistochemical co-staining of microvessel marker CD31 and cellular proliferation marker BrdU 21 days after ischemia showed an impaired angiogenic activity in the TNFR-1-/- mice. These data were supported by Western blot analysis, which indicated a decreased expression of angiopoietin-1 (Ang-1) and its receptor Tie-2 in TNFR-1-/- mice. Our results suggest that a deficiency in TNFR-1 prevents the activation of death-related proteins downstream to TNF-alpha and attenuates apoptosis in acute limb ischemia, but the lack of TNFR-1 signaling hinders the belated angiogenesis mediated by the Ang-1/Tie-2 pathway.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amputation, Surgical
  • Angiopoietin-1 / metabolism
  • Animals
  • Apoptosis
  • Caspase 3 / metabolism
  • Disease Models, Animal
  • Female
  • Ischemia / metabolism*
  • Ischemia / pathology
  • Ischemia / physiopathology
  • Ischemia / surgery
  • Laser-Doppler Flowmetry
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Muscle, Skeletal / blood supply*
  • Muscle, Skeletal / metabolism
  • Muscle, Skeletal / pathology
  • Neovascularization, Physiologic*
  • Receptor, TIE-2 / metabolism
  • Receptors, Tumor Necrosis Factor, Type I / deficiency
  • Receptors, Tumor Necrosis Factor, Type I / genetics
  • Receptors, Tumor Necrosis Factor, Type I / metabolism*
  • Regional Blood Flow
  • Signal Transduction*
  • Time Factors
  • Tumor Necrosis Factor-alpha / metabolism*
  • bcl-2-Associated X Protein / metabolism

Substances

  • Angiopoietin-1
  • Bax protein, mouse
  • Receptors, Tumor Necrosis Factor, Type I
  • Tnfrsf1a protein, mouse
  • Tumor Necrosis Factor-alpha
  • bcl-2-Associated X Protein
  • Receptor, TIE-2
  • Casp3 protein, mouse
  • Caspase 3