The role of the cyclooxygenase-2 pathway in tissue ischemia and revascularization following skeletal muscle injury induced by bothropic snake venom

Microvasc Res. 2025 Jan:157:104760. doi: 10.1016/j.mvr.2024.104760. Epub 2024 Nov 5.

Abstract

Bothrops asper venom (Bav) contains metalloproteinases that disrupt the microvascular system, impairing muscle tissue regeneration after injury. This study investigated the impact of the cyclooxygenase-2 (COX-2) pathway on vascular injury and revascularization in muscle injuries induced by Bav. Mice were injected with Bav into the gastrocnemius muscle and treated with lumiracoxib, a selective COX-2 inhibitor, 30 min, 2 days, and 6 days post-Bav injection. Muscle tissue was analyzed at 24 h, 7 days, and 21 days post-injection. A decrease in COX-2 expression at 24 h post-Bav injection indicated significant necrosis and tissue loss. Both Bav injection and lumiracoxib treatment influenced the decrease of prostaglandin (PG)D2 and PGE2 production. Seven and 21 days post-Bav injections, COX-2 expression increased, along with PGDs levels unaffected by lumiracoxib, indicating that the other isoform COX-1 pathway could contribute to the release of PGs. Bav/lumiracoxib treated animals presented exacerbated limb ischemia, implying that COX-2-derived prostaglandins preserve vessel integrity. CD31, an angiogenesis marker, initially (24 h) decreased post-Bav injection but increased at 7 and 21 days in Bav/lumiracoxib mice, suggesting a down-modulatory role for COX-2-derived prostaglandins in early angiogenesis and tissue regeneration. Vascular endothelial growth factor (VEGF) production rose 7 days post-Bav injection, supporting its role in angiogenesis. Previous treatment with lumiracoxib promoted release of VEGF levels 21 days post-Bav injury showing that the inhibition of COX-2 pathway in the early stage of revascularization stimulates the neovascularization regulated by elevated release of VEGF. Similarly, metalloproteinases (MMPs), such as MMP-9, MMP-10, and MMP-13, crucial for vascular remodeling, were elevated 21 days after Bav/lumiracoxib treatment. In conclusion, the COX-2 pathway is essential to decrease the high grade of ischemia caused by acute injury induced by Bav. However, the decrease of activity in the COX-2 pathway in the first stages of revascularization contributes to the elevated production of key pro-angiogenic mediators that up-regulate the restoration of microvasculature and blood flow in muscle tissue injured by botropic venoms.

Keywords: Bothrops venom myotoxicity; Cyclooxygenase-2; Revascularization.

MeSH terms

  • Animals
  • Bothrops*
  • Crotalid Venoms* / toxicity
  • Cyclooxygenase 2 Inhibitors* / pharmacology
  • Cyclooxygenase 2* / metabolism
  • Dinoprostone / metabolism
  • Disease Models, Animal*
  • Hindlimb / blood supply
  • Ischemia* / drug therapy
  • Ischemia* / metabolism
  • Ischemia* / pathology
  • Ischemia* / physiopathology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Muscle, Skeletal* / blood supply
  • Muscle, Skeletal* / drug effects
  • Muscle, Skeletal* / metabolism
  • Neovascularization, Physiologic* / drug effects
  • Platelet Endothelial Cell Adhesion Molecule-1 / metabolism
  • Regional Blood Flow
  • Signal Transduction*
  • Time Factors
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Cyclooxygenase 2
  • Crotalid Venoms
  • Cyclooxygenase 2 Inhibitors
  • Ptgs2 protein, mouse
  • vascular endothelial growth factor A, mouse
  • Platelet Endothelial Cell Adhesion Molecule-1
  • Vascular Endothelial Growth Factor A
  • Pecam1 protein, mouse
  • Dinoprostone