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 Fe 2+-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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 29223-29230 |
| Number of pages | 8 |
| Journal | Journal of Biological Chemistry |
| Volume | 271 |
| Issue number | 46 |
| DOIs | |
| State | Published - 1996 |
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ASJC Scopus subject areas
- Biochemistry
Cite this
Measurement of nitric oxide and peroxynitrite generation in the postischemic heart : Evidence for peroxynitrite-mediated reperfusion injury. / Wang, Penghai; Zweier, Jay L.
In: Journal of Biological Chemistry, Vol. 271, No. 46, 1996, p. 29223-29230.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Measurement of nitric oxide and peroxynitrite generation in the postischemic heart
T2 - Evidence for peroxynitrite-mediated reperfusion injury
AU - Wang, Penghai
AU - Zweier, Jay L.
PY - 1996
Y1 - 1996
N2 - 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 Fe 2+-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.
AB - 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 Fe 2+-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.
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U2 - 10.1074/jbc.271.46.29223
DO - 10.1074/jbc.271.46.29223
M3 - Article
VL - 271
SP - 29223
EP - 29230
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 46
ER -