Evaluation of systemic blood NO dynamics by EPR spectroscopy: HbNO as an endogenous index of NO
Departments of 1 Pharmacology and 2 Integrative Physiology, The University of Tokushima School of Medicine, Tokushima 770-8503, Japan Submitted 10 December 2002 ; accepted in final form 19 March 2003 The measurement of hemoglobin-nitric oxide (NO) adduct (HbNO) in whole blood by the electron paramag...
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| Veröffentlicht in: | American journal of physiology. Heart and circulatory physiology 2003-08, Vol.285 (2), p.H589-H596 |
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| container_title | American journal of physiology. Heart and circulatory physiology |
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| creator | Kirima, Kazuyoshi Tsuchiya, Koichiro Sei, Hiroyoshi Hasegawa, Toyoshi Shikishima, Michiyo Motobayashi, Yuki Morita, Kyoji Yoshizumi, Masanori Tamaki, Toshiaki |
| description | Departments of 1 Pharmacology and
2 Integrative Physiology, The University of Tokushima
School of Medicine, Tokushima 770-8503, Japan
Submitted 10 December 2002
; accepted in final form 19 March 2003
The measurement of hemoglobin-nitric oxide (NO) adduct (HbNO) in whole
blood by the electron paramagnetic resonance (EPR) method seems relevant for
the assessment of systemic NO levels. However, ceruloplasmin and unknown
radical species overlap the same magnetic field as that of HbNO. To reveal the
EPR spectrum of HbNO, we then introduced the EPR signal subtraction method,
which is based on the computer-assisted subtraction of the digitized EPR
spectrum of HbNO-depleted blood from that of sample blood using the software.
Rats were treated with
N -nitro- L -arginine methyl ester
( L -NAME; 120 mg · kg 1 ·
day 1 ) for 1 wk to obtain HbNO-depleted blood.
When this method was applied to the analysis of untreated fresh whole blood,
the five-coordinate state of HbNO was observed. HbNO concentration in
pentobarbital-anesthetized rats was augmented (change in [HbNO] =
1.65.5 µM) by infusion of L -arginine (0.20.6 g/kg)
but not D -arginine. Using this method, we attempted to evaluate the
effects of temocapril on HbNO dynamics in an L -NAME-induced rat
endothelial dysfunction model. The oral administration of L -NAME
for 2 wk induced a serious hypertension, and the HbNO concentration was
reduced (change in [HbNO] = 5.7 µM). Coadministration of temocapril dose
dependently improved both changes in blood pressure and the systemic HbNO
concentration. In this study, we succeeded in measuring the blood HbNO level
as an index of NO by the EPR HbNO signal subtraction method. We also
demonstrated that temocapril improves abnormalities of NO dynamics in
L -NAME-induced endothelial dysfunction rats using the EPR HbNO
signal subtraction method.
nitric oxide; hemoglobin-nitric oxide adduct; electron paramagnetic resonance; temocapril
Address for reprint requests and other correspondence: T. Tamaki, Dept. of
Pharmacology, The Univ. of Tokushima School of Medicine, 3-18-15 Kuramoto,
Tokushima 770-8503, Japan (E-mail:
tamaki{at}basic.med.tokushima-u.ac.jp ). |
| doi_str_mv | 10.1152/ajpheart.01010.2002 |
| format | Article |
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2 Integrative Physiology, The University of Tokushima
School of Medicine, Tokushima 770-8503, Japan
Submitted 10 December 2002
; accepted in final form 19 March 2003
The measurement of hemoglobin-nitric oxide (NO) adduct (HbNO) in whole
blood by the electron paramagnetic resonance (EPR) method seems relevant for
the assessment of systemic NO levels. However, ceruloplasmin and unknown
radical species overlap the same magnetic field as that of HbNO. To reveal the
EPR spectrum of HbNO, we then introduced the EPR signal subtraction method,
which is based on the computer-assisted subtraction of the digitized EPR
spectrum of HbNO-depleted blood from that of sample blood using the software.
Rats were treated with
N -nitro- L -arginine methyl ester
( L -NAME; 120 mg · kg 1 ·
day 1 ) for 1 wk to obtain HbNO-depleted blood.
When this method was applied to the analysis of untreated fresh whole blood,
the five-coordinate state of HbNO was observed. HbNO concentration in
pentobarbital-anesthetized rats was augmented (change in [HbNO] =
1.65.5 µM) by infusion of L -arginine (0.20.6 g/kg)
but not D -arginine. Using this method, we attempted to evaluate the
effects of temocapril on HbNO dynamics in an L -NAME-induced rat
endothelial dysfunction model. The oral administration of L -NAME
for 2 wk induced a serious hypertension, and the HbNO concentration was
reduced (change in [HbNO] = 5.7 µM). Coadministration of temocapril dose
dependently improved both changes in blood pressure and the systemic HbNO
concentration. In this study, we succeeded in measuring the blood HbNO level
as an index of NO by the EPR HbNO signal subtraction method. We also
demonstrated that temocapril improves abnormalities of NO dynamics in
L -NAME-induced endothelial dysfunction rats using the EPR HbNO
signal subtraction method.
nitric oxide; hemoglobin-nitric oxide adduct; electron paramagnetic resonance; temocapril
Address for reprint requests and other correspondence: T. Tamaki, Dept. of
Pharmacology, The Univ. of Tokushima School of Medicine, 3-18-15 Kuramoto,
Tokushima 770-8503, Japan (E-mail:
tamaki{at}basic.med.tokushima-u.ac.jp ).</description><identifier>ISSN: 0363-6135</identifier><identifier>EISSN: 1522-1539</identifier><identifier>DOI: 10.1152/ajpheart.01010.2002</identifier><identifier>PMID: 12663263</identifier><language>eng</language><publisher>United States</publisher><subject>Angiotensin-Converting Enzyme Inhibitors - pharmacology ; Animals ; Arginine - pharmacology ; Calibration ; Electron Spin Resonance Spectroscopy - methods ; Enzyme Inhibitors - pharmacology ; Hemoglobins - metabolism ; Hypertension - chemically induced ; Hypertension - metabolism ; Male ; NG-Nitroarginine Methyl Ester - pharmacology ; Nitric Oxide - blood ; Rats ; Rats, Sprague-Dawley ; Thiazepines - pharmacology</subject><ispartof>American journal of physiology. Heart and circulatory physiology, 2003-08, Vol.285 (2), p.H589-H596</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c391t-c98d8b5c914dfa2bb36d3b0fcc8902898b20429bb6e7a815297a968269a9e15b3</citedby><cites>FETCH-LOGICAL-c391t-c98d8b5c914dfa2bb36d3b0fcc8902898b20429bb6e7a815297a968269a9e15b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,3026,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12663263$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kirima, Kazuyoshi</creatorcontrib><creatorcontrib>Tsuchiya, Koichiro</creatorcontrib><creatorcontrib>Sei, Hiroyoshi</creatorcontrib><creatorcontrib>Hasegawa, Toyoshi</creatorcontrib><creatorcontrib>Shikishima, Michiyo</creatorcontrib><creatorcontrib>Motobayashi, Yuki</creatorcontrib><creatorcontrib>Morita, Kyoji</creatorcontrib><creatorcontrib>Yoshizumi, Masanori</creatorcontrib><creatorcontrib>Tamaki, Toshiaki</creatorcontrib><title>Evaluation of systemic blood NO dynamics by EPR spectroscopy: HbNO as an endogenous index of NO</title><title>American journal of physiology. Heart and circulatory physiology</title><addtitle>Am J Physiol Heart Circ Physiol</addtitle><description>Departments of 1 Pharmacology and
2 Integrative Physiology, The University of Tokushima
School of Medicine, Tokushima 770-8503, Japan
Submitted 10 December 2002
; accepted in final form 19 March 2003
The measurement of hemoglobin-nitric oxide (NO) adduct (HbNO) in whole
blood by the electron paramagnetic resonance (EPR) method seems relevant for
the assessment of systemic NO levels. However, ceruloplasmin and unknown
radical species overlap the same magnetic field as that of HbNO. To reveal the
EPR spectrum of HbNO, we then introduced the EPR signal subtraction method,
which is based on the computer-assisted subtraction of the digitized EPR
spectrum of HbNO-depleted blood from that of sample blood using the software.
Rats were treated with
N -nitro- L -arginine methyl ester
( L -NAME; 120 mg · kg 1 ·
day 1 ) for 1 wk to obtain HbNO-depleted blood.
When this method was applied to the analysis of untreated fresh whole blood,
the five-coordinate state of HbNO was observed. HbNO concentration in
pentobarbital-anesthetized rats was augmented (change in [HbNO] =
1.65.5 µM) by infusion of L -arginine (0.20.6 g/kg)
but not D -arginine. Using this method, we attempted to evaluate the
effects of temocapril on HbNO dynamics in an L -NAME-induced rat
endothelial dysfunction model. The oral administration of L -NAME
for 2 wk induced a serious hypertension, and the HbNO concentration was
reduced (change in [HbNO] = 5.7 µM). Coadministration of temocapril dose
dependently improved both changes in blood pressure and the systemic HbNO
concentration. In this study, we succeeded in measuring the blood HbNO level
as an index of NO by the EPR HbNO signal subtraction method. We also
demonstrated that temocapril improves abnormalities of NO dynamics in
L -NAME-induced endothelial dysfunction rats using the EPR HbNO
signal subtraction method.
nitric oxide; hemoglobin-nitric oxide adduct; electron paramagnetic resonance; temocapril
Address for reprint requests and other correspondence: T. Tamaki, Dept. of
Pharmacology, The Univ. of Tokushima School of Medicine, 3-18-15 Kuramoto,
Tokushima 770-8503, Japan (E-mail:
tamaki{at}basic.med.tokushima-u.ac.jp ).</description><subject>Angiotensin-Converting Enzyme Inhibitors - pharmacology</subject><subject>Animals</subject><subject>Arginine - pharmacology</subject><subject>Calibration</subject><subject>Electron Spin Resonance Spectroscopy - methods</subject><subject>Enzyme Inhibitors - pharmacology</subject><subject>Hemoglobins - metabolism</subject><subject>Hypertension - chemically induced</subject><subject>Hypertension - metabolism</subject><subject>Male</subject><subject>NG-Nitroarginine Methyl Ester - pharmacology</subject><subject>Nitric Oxide - blood</subject><subject>Rats</subject><subject>Rats, Sprague-Dawley</subject><subject>Thiazepines - pharmacology</subject><issn>0363-6135</issn><issn>1522-1539</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kFtPwjAYhhujETz8AhPTPzDswZWWO0NATAgYo9dNT4ORsS7rpuzfWwTlyvSiSfs-b77vAeAOowHGKXlQm2rtVN0MEI5nQBAiZ6Aff0iCUyrOQR9RRhOGadoDVyFsEELpkNFL0MOEMUoY7QM5-VRFq5rcl9BnMHShcdvcQF14b-FiCW1XqvgQoO7g5PUNhsqZpvbB-KobwZmOERWgKqErrV-50rcB5qV1u33dYnkDLjJVBHd7vK_Bx3TyPp4l8-Xzy_hpnhgqcJMYwS3XqRH40WaKaE2ZpRplxnCBCBdcE_RIhNbMDRWPO4qhEowTJpRwONX0GtBDr4mzhdplsqrzrao7iZHc65K_uuSPLrnXFan7A1W1euvsiTn6iYHRIbDOV-uvvHayWnch94VfdXLaFsW72zV_1YSnkshZyoWsbBbhwf_w3zgniH4Di7mN9Q</recordid><startdate>20030801</startdate><enddate>20030801</enddate><creator>Kirima, Kazuyoshi</creator><creator>Tsuchiya, Koichiro</creator><creator>Sei, Hiroyoshi</creator><creator>Hasegawa, Toyoshi</creator><creator>Shikishima, Michiyo</creator><creator>Motobayashi, Yuki</creator><creator>Morita, Kyoji</creator><creator>Yoshizumi, Masanori</creator><creator>Tamaki, Toshiaki</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope></search><sort><creationdate>20030801</creationdate><title>Evaluation of systemic blood NO dynamics by EPR spectroscopy: HbNO as an endogenous index of NO</title><author>Kirima, Kazuyoshi ; Tsuchiya, Koichiro ; Sei, Hiroyoshi ; Hasegawa, Toyoshi ; Shikishima, Michiyo ; Motobayashi, Yuki ; Morita, Kyoji ; Yoshizumi, Masanori ; Tamaki, Toshiaki</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c391t-c98d8b5c914dfa2bb36d3b0fcc8902898b20429bb6e7a815297a968269a9e15b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Angiotensin-Converting Enzyme Inhibitors - pharmacology</topic><topic>Animals</topic><topic>Arginine - pharmacology</topic><topic>Calibration</topic><topic>Electron Spin Resonance Spectroscopy - methods</topic><topic>Enzyme Inhibitors - pharmacology</topic><topic>Hemoglobins - metabolism</topic><topic>Hypertension - chemically induced</topic><topic>Hypertension - metabolism</topic><topic>Male</topic><topic>NG-Nitroarginine Methyl Ester - pharmacology</topic><topic>Nitric Oxide - blood</topic><topic>Rats</topic><topic>Rats, Sprague-Dawley</topic><topic>Thiazepines - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kirima, Kazuyoshi</creatorcontrib><creatorcontrib>Tsuchiya, Koichiro</creatorcontrib><creatorcontrib>Sei, Hiroyoshi</creatorcontrib><creatorcontrib>Hasegawa, Toyoshi</creatorcontrib><creatorcontrib>Shikishima, Michiyo</creatorcontrib><creatorcontrib>Motobayashi, Yuki</creatorcontrib><creatorcontrib>Morita, Kyoji</creatorcontrib><creatorcontrib>Yoshizumi, Masanori</creatorcontrib><creatorcontrib>Tamaki, Toshiaki</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><jtitle>American journal of physiology. Heart and circulatory physiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kirima, Kazuyoshi</au><au>Tsuchiya, Koichiro</au><au>Sei, Hiroyoshi</au><au>Hasegawa, Toyoshi</au><au>Shikishima, Michiyo</au><au>Motobayashi, Yuki</au><au>Morita, Kyoji</au><au>Yoshizumi, Masanori</au><au>Tamaki, Toshiaki</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Evaluation of systemic blood NO dynamics by EPR spectroscopy: HbNO as an endogenous index of NO</atitle><jtitle>American journal of physiology. Heart and circulatory physiology</jtitle><addtitle>Am J Physiol Heart Circ Physiol</addtitle><date>2003-08-01</date><risdate>2003</risdate><volume>285</volume><issue>2</issue><spage>H589</spage><epage>H596</epage><pages>H589-H596</pages><issn>0363-6135</issn><eissn>1522-1539</eissn><abstract>Departments of 1 Pharmacology and
2 Integrative Physiology, The University of Tokushima
School of Medicine, Tokushima 770-8503, Japan
Submitted 10 December 2002
; accepted in final form 19 March 2003
The measurement of hemoglobin-nitric oxide (NO) adduct (HbNO) in whole
blood by the electron paramagnetic resonance (EPR) method seems relevant for
the assessment of systemic NO levels. However, ceruloplasmin and unknown
radical species overlap the same magnetic field as that of HbNO. To reveal the
EPR spectrum of HbNO, we then introduced the EPR signal subtraction method,
which is based on the computer-assisted subtraction of the digitized EPR
spectrum of HbNO-depleted blood from that of sample blood using the software.
Rats were treated with
N -nitro- L -arginine methyl ester
( L -NAME; 120 mg · kg 1 ·
day 1 ) for 1 wk to obtain HbNO-depleted blood.
When this method was applied to the analysis of untreated fresh whole blood,
the five-coordinate state of HbNO was observed. HbNO concentration in
pentobarbital-anesthetized rats was augmented (change in [HbNO] =
1.65.5 µM) by infusion of L -arginine (0.20.6 g/kg)
but not D -arginine. Using this method, we attempted to evaluate the
effects of temocapril on HbNO dynamics in an L -NAME-induced rat
endothelial dysfunction model. The oral administration of L -NAME
for 2 wk induced a serious hypertension, and the HbNO concentration was
reduced (change in [HbNO] = 5.7 µM). Coadministration of temocapril dose
dependently improved both changes in blood pressure and the systemic HbNO
concentration. In this study, we succeeded in measuring the blood HbNO level
as an index of NO by the EPR HbNO signal subtraction method. We also
demonstrated that temocapril improves abnormalities of NO dynamics in
L -NAME-induced endothelial dysfunction rats using the EPR HbNO
signal subtraction method.
nitric oxide; hemoglobin-nitric oxide adduct; electron paramagnetic resonance; temocapril
Address for reprint requests and other correspondence: T. Tamaki, Dept. of
Pharmacology, The Univ. of Tokushima School of Medicine, 3-18-15 Kuramoto,
Tokushima 770-8503, Japan (E-mail:
tamaki{at}basic.med.tokushima-u.ac.jp ).</abstract><cop>United States</cop><pmid>12663263</pmid><doi>10.1152/ajpheart.01010.2002</doi></addata></record> |
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| identifier | ISSN: 0363-6135 |
| ispartof | American journal of physiology. Heart and circulatory physiology, 2003-08, Vol.285 (2), p.H589-H596 |
| issn | 0363-6135 1522-1539 |
| language | eng |
| recordid | cdi_crossref_primary_10_1152_ajpheart_01010_2002 |
| source | MEDLINE; American Physiological Society; EZB-FREE-00999 freely available EZB journals |
| subjects | Angiotensin-Converting Enzyme Inhibitors - pharmacology Animals Arginine - pharmacology Calibration Electron Spin Resonance Spectroscopy - methods Enzyme Inhibitors - pharmacology Hemoglobins - metabolism Hypertension - chemically induced Hypertension - metabolism Male NG-Nitroarginine Methyl Ester - pharmacology Nitric Oxide - blood Rats Rats, Sprague-Dawley Thiazepines - pharmacology |
| title | Evaluation of systemic blood NO dynamics by EPR spectroscopy: HbNO as an endogenous index of NO |
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