Treatment of stroke with (Z)-1-[N-(2-aminoethyl)-N-(2-ammonioethyl) amino] diazen-1-ium-1, 2-diolate and bone marrow stromal cells upregulates angiopoietin-1/Tie2 and enhances neovascularization.

Abstract

Neovascularization may contribute to functional recovery after neural injury. Combination treatment of stroke with a nitric oxide donor, (Z)-1-[N-(2-aminoethyl)-N-(2-ammonioethyl) amino] diazen-1-ium-1, 2-diolate (DETA-NONOate) and bone marrow stromal cells promotes functional recovery. However, the mechanisms underlying functional improvement have not been elucidated. In this study, we tested the hypothesis that combination treatment upregulates angiopoietin-1 and its receptor Tie2 in the ischemic brain and bone marrow stromal cells, thereby enhancing cerebral neovascularization after stroke. Adult wild type male C57BL/6 mice were i.v. administered PBS, bone marrow stromal cells 5x10(5), DETA-NONOate 0.4 mg/kg or combination DETA-NONOate with bone marrow stromal cells (n=12/group) after middle cerebral artery occlusion. Combination treatment significantly upregulated angiopoietin-1/Tie2 and tight junction protein (occludin) expression, and increased the number, diameter and perimeter of blood vessels in the ischemic brain compared with vehicle control (mean+ or -S.E., P<0.05). In vitro, DETA-NONOate significantly increased angiopoietin-1/Tie2 protein (n=6/group) and Tie2 mRNA (n=3/group) expression in bone marrow stromal cells. DETA-NONOate also significantly increased angiopoietin-1 protein (n=6/group) and mRNA (n=3/group) expression in mouse brain endothelial cells (P<0.05). Angiopoietin-1 mRNA (n=3/group) was significantly increased in mouse brain endothelial cells treated with DETA-NONOate in combination with bone marrow stromal cell-conditioned medium compared with cells treated with bone marrow stromal cell-conditioned medium or DETA-NONOate alone. Mouse brain endothelial cell capillary tube-like formation assays (n=6/group) showed that angiopoietin-1 peptide, the supernatant of bone marrow stromal cells and DETA-NONOate significantly increased capillary tube formation compared with vehicle control. Combination treatment significantly increased capillary tube formation compared with DETA-NONOate treatment alone. Inhibition of angiopoietin-1 significantly attenuated combination treatment-induced tube formation. Our data indicated that combination treatment of stroke with DETA-NONOate and bone marrow stromal cells promotes neovascularization, which is at least partially mediated by upregulation of the angiopoietin-1/Tie2 axis.

• Authors: X Cui, J Chen, A Zacharek, C Roberts, S Savant-Bhonsale, M Chopp
• Journal: Neuroscience (Neuroscience) Vol. 156 Issue 1 Pg. 155-64 (Sep 22 2008) ISSN: 0306-4522 [Print] United States
• PMID: 18691637 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Angiopoietin-1
• Angpt1 protein, mouse
• Nitric Oxide Donors
• Nitroso Compounds
• RNA, Messenger
• 2,2'-(hydroxynitrosohydrazono)bis-ethanamine
• Receptor, TIE-2

Topics

• Angiopoietin-1 (agonists, antagonists & inhibitors, metabolism)
• Animals
• Bone Marrow Transplantation (methods)
• Capillaries (drug effects, metabolism)
• Cells, Cultured
• Cerebral Arteries (drug effects, metabolism)
• Coculture Techniques
• Disease Models, Animal
• Endothelium, Vascular (drug effects, metabolism)
• Male
• Mice
• Mice, Inbred C57BL
• Neovascularization, Physiologic (drug effects, physiology)
• Nitric Oxide Donors (pharmacology, therapeutic use)
• Nitroso Compounds (pharmacology, therapeutic use)
• RNA, Messenger (drug effects, metabolism)
• Receptor, TIE-2 (genetics)
• Regeneration (drug effects, physiology)
• Stroke (drug therapy, surgery)
• Stromal Cells (transplantation)
• Tight Junctions (drug effects, metabolism)
• Up-Regulation (drug effects, physiology)