Abstract:
OBJECTIVE:Research in the pathophysiology of ischemia/reperfusion or redox signaling is hindered by lack of simple methodology to measure short-lived oxygen radicals. In the presence of hydroxyl radical ((*)OH), d-phenylalanine (d-Phe) yields para-, meta- and ortho-tyrosine. We have previously demonstrated that d-Phe can accurately detect (*)OH formation in chemical, enzymatic and cellular systems by simple HPLC methodology [Anal Biochem 290:138;2001]. In the present study, we tested whether d-Phe hydroxylation can be used to detect (*)OH formation in intact organs. METHODS:Rat hearts were perfused with buffer containing 5 mM d-Phe and subjected to 30 min of total global ischemia at 37 degrees C followed by 45 min of reperfusion. Quantitative analysis of the three hydroxytyrosine isomers was achieved by HPLC-based electrochemical detection of cardiac venous effluent, with the analytical cells operating in the oxidative mode. The detection limit of this assay was <10 fmol. RESULTS:Under baseline conditions, hydroxytyrosine release from the heart was very low ( congruent with0.8 nmol/min/g). However, a prominent tyrosine burst occurred immediately upon post-ischemic reflow. In cardiac effluent collected 40 s into reperfusion, the hydroxytyrosine concentration was more than 40 times greater than at baseline; hydroxytyrosine concentration then progressively declined, to return to pre-ischemic values by 5 min of reperfusion. In parallel experiments, formation of hydroxytyrosines was markedly reduced in hearts reperfused in the presence of the (*)OH scavenger mannitol. Inclusion of 5 mm d-Phe in the perfusion medium altered neither basal cardiac function nor coronary vascular tone, but it enhanced recovery of myocardial function during post-ischemic reperfusion, consistent with direct reaction with (*)OH. CONCLUSION:Our results demonstrate that d-Phe is a sensitive method for detection of (*)OH generation in the heart. Since d-Phe is not a substrate for endogenous enzymes, it can be exploited as a reliable method to measure (*)OH formation under a variety of pathophysiological conditions.
journal_name
Cardiovasc Resjournal_title
Cardiovascular researchauthors
Biondi R,Ambrosio G,Liebgott T,Cardounel AJ,Bettini M,Tritto I,Zweier JLdoi
10.1016/j.cardiores.2006.03.005subject
Has Abstractpub_date
2006-07-15 00:00:00pages
322-30issue
2eissn
0008-6363issn
1755-3245pii
S0008-6363(06)00119-2journal_volume
71pub_type
杂志文章abstract:OBJECTIVE:The pulmonary circulation may contribute to elevated plasma levels of endothelin-1 (ET-1) in congestive heart failure (CHF). The aims of the present study were to determine the mechanisms of increased secretion of ET-1 from the pulmonary circulation in CHF. METHODS:Juvenile pigs were subjected to sham operat...
journal_title:Cardiovascular research
pub_type: 杂志文章
doi:10.1016/j.cardiores.2004.03.016
更新日期:2004-07-01 00:00:00
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doi:10.1093/cvr/cvx236
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journal_title:Cardiovascular research
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doi:10.1016/s0008-6363(01)00278-4
更新日期:2001-07-01 00:00:00
abstract::The significance of pre-existing coronary collaterals for preservation of the myocardium during coronary occlusion and reperfusion was studied in anaesthetised open-chest dogs. The extent of myocardial ischaemic damage was assessed by the accumulation of calcium in the central zone of the ischaemic area and the size o...
journal_title:Cardiovascular research
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doi:10.1093/cvr/14.5.288
更新日期:1980-05-01 00:00:00
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pub_type: 杂志文章
doi:10.1016/s0008-6363(99)00045-0
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journal_title:Cardiovascular research
pub_type: 杂志文章,随机对照试验
doi:10.1093/cvr/cvy227
更新日期:2019-03-01 00:00:00
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journal_title:Cardiovascular research
pub_type: 杂志文章
doi:10.1093/cvr/27.7.1359
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journal_title:Cardiovascular research
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doi:10.1093/cvr/25.10.869
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journal_title:Cardiovascular research
pub_type: 杂志文章
doi:
更新日期:1996-02-01 00:00:00
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journal_title:Cardiovascular research
pub_type: 杂志文章
doi:10.1016/j.cardiores.2004.06.028
更新日期:2005-05-01 00:00:00
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journal_title:Cardiovascular research
pub_type: 杂志文章
doi:10.1093/cvr/cvaa239
更新日期:2020-08-10 00:00:00
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pub_type: 杂志文章,评审
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更新日期:1992-01-01 00:00:00
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更新日期:2020-04-28 00:00:00
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更新日期:2009-08-01 00:00:00
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journal_title:Cardiovascular research
pub_type: 杂志文章
doi:10.1093/cvr/cvy105
更新日期:2018-09-01 00:00:00
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journal_title:Cardiovascular research
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doi:10.1093/cvr/20.11.816
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journal_title:Cardiovascular research
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doi:10.1093/cvr/27.12.2135
更新日期:1993-12-01 00:00:00
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更新日期:2017-01-01 00:00:00
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更新日期:2000-09-01 00:00:00
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journal_title:Cardiovascular research
pub_type: 杂志文章
doi:10.1093/cvr/27.3.494
更新日期:1993-03-01 00:00:00
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journal_title:Cardiovascular research
pub_type: 杂志文章
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更新日期:2003-09-01 00:00:00
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journal_title:Cardiovascular research
pub_type: 杂志文章
doi:
更新日期:1996-02-01 00:00:00
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journal_title:Cardiovascular research
pub_type: 杂志文章
doi:10.1016/s0008-6363(02)00657-0
更新日期:2003-01-01 00:00:00
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journal_title:Cardiovascular research
pub_type: 杂志文章
doi:10.1016/s0008-6363(00)00013-4
更新日期:2000-04-01 00:00:00
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journal_title:Cardiovascular research
pub_type: 杂志文章,评审
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更新日期:2004-05-01 00:00:00
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journal_title:Cardiovascular research
pub_type: 杂志文章
doi:10.1016/s0008-6363(03)00353-5
更新日期:2003-07-01 00:00:00
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pub_type: 杂志文章
doi:10.1016/s0008-6363(97)00295-2
更新日期:1998-03-01 00:00:00