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Measurement of nitric oxide and peroxynitrite generation in the postischemic heart. Evidence for peroxynitrite-mediated reperfusion injury. | ||
| Authors: | Wang, P and Zweier, J L | |
| Citation: | J. Biol. Chem., 271: 29223-30 (1996) | |
| Pub Med ID: | 8910581 | |
| Year: | 1996 | |
| Abstract: | Altered nitric oxide (NO.) production is a critical factor in tissue reperfusion injury; however, controversy remains regarding these alterations and how they cause injury. Since superoxide (O-2) generation is triggered during the early period of reperfusion the cytotoxic oxidant peroxynitrite (ONOO-) could be formed, but it is not known if this occurs. Therefore electron paramagnetic resonance and chemiluminescence studies were performed of the magnitude and time course of NO., O-2, and ONOO- formation in the postischemic heart. Isolated rat hearts were subjected either to normal perfusion or to reperfusion after 30 min of ischemia in the presence of the NO. trap Fe2+-N-methyl-D-glucamine dithiocarbamate with electron paramagnetic resonance measurements performed on the effluent. Although only trace signals were present prior to ischemia, prominent NO. adduct signals were seen during the first 2 min of reflow which were abolished by nitric oxide synthase (NOS) inhibition. Similar studies with the O-2 trap 5, 5-dimethyl-1-pyrroline N-oxide demonstrated a burst of O-2 generation over the first 2 min of reflow. Chemiluminescence measurements using 5-amino-2,3-dihydro-1,4-phthalazinedione (luminol) demonstrated a similar marked increase in ONOO- which was blocked by NOS inhibitors or superoxide dismutase. NOS inhibition or superoxide dismutase greatly enhanced the recovery of contractile function in postischemic hearts. Immunohistology demonstrated that the ONOO--mediated nitration product nitrotyrosine was formed in postischemic hearts but not in normally perfused controls. Thus, NO. formation is increased during the early period of reflow and reacts with O-2 to form ONOO-, which results in amino acid nitration and cellular injury. | |
| Application: | Immunohistochemistry (tissue) | |
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