Reticulocyte 15-Lipoxygenase-I Is Important in Acetylcholine-Induced Endothelium-Dependent Vasorelaxation in Rabbit Aorta
OBJECTIVE—Aortic 15-lipoxygenase (15-LO) metabolizes arachidonic acid (AA) to 15-hydroperoxyeicosatetraenoic acid, which is then converted to the vasodilators 15-hydroxy-11,12-epoxyeicosatrienoic acid and 11,12,15-trihydroxyeicosatrienoic acid. These metabolites contribute to endothelium-dependent r...
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| Veröffentlicht in: | Arteriosclerosis, thrombosis, and vascular biology thrombosis, and vascular biology, 2006-01, Vol.26 (1), p.78-84 |
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| creator | Tang, Xin Holmes, Blythe B Nithipatikom, Kasem Hillard, Cecilia J Kuhn, Hartmut Campbell, William B |
| description | OBJECTIVE—Aortic 15-lipoxygenase (15-LO) metabolizes arachidonic acid (AA) to 15-hydroperoxyeicosatetraenoic acid, which is then converted to the vasodilators 15-hydroxy-11,12-epoxyeicosatrienoic acid and 11,12,15-trihydroxyeicosatrienoic acid. These metabolites contribute to endothelium-dependent relaxations of rabbit aorta to AA and acetylcholine. We investigated the identity of rabbit aortic 15-LO and studied its importance in the regulation of vascular tone.
METHODS AND RESULTS—RT-PCR using 12-lipoxygenase/15-LO specific primers resulted in a 572-bp product with a sequence identical to 15-LO-I from rabbit aorta. A RT-PCR/restriction digest strategy excluded expression of 12-lipoxygenase. Immunoblotting revealed 15-LO-I expression in rabbit endothelial and smooth muscle cells. Aortic homogenates and cytosolic fractions metabolize AA to 15(S)-hydroxyeicosatetraenoic acid and linoleic acid to 13(S)-hydroxyoctadecadienoic acid. This activity was blocked by LO inhibitors. The kinetic characteristics (Michaelis constant of aortic 15-LO is 2.2±0.3 μmol/L for AA and 23.5±3.3 μmol/L for linoleic acid) of aortic 15-LO were similar to those of the purified 15-LO-I. An antisense oligonucleotide inhibited 15-LO-I expression in rabbit aorta. Indomethacin and nitro-l-arginine-resistant relaxations to acetylcholine were inhibited by 15-LO-I antisense oligonucleotide but not by the scrambled oligonucleotide.
CONCLUSIONS—15-LO-I is expressed in rabbit aortic endothelium and is important in endothelium-dependent regulation of vascular tone. |
| doi_str_mv | 10.1161/01.ATV.0000191640.73313.ad |
| format | Article |
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METHODS AND RESULTS—RT-PCR using 12-lipoxygenase/15-LO specific primers resulted in a 572-bp product with a sequence identical to 15-LO-I from rabbit aorta. A RT-PCR/restriction digest strategy excluded expression of 12-lipoxygenase. Immunoblotting revealed 15-LO-I expression in rabbit endothelial and smooth muscle cells. Aortic homogenates and cytosolic fractions metabolize AA to 15(S)-hydroxyeicosatetraenoic acid and linoleic acid to 13(S)-hydroxyoctadecadienoic acid. This activity was blocked by LO inhibitors. The kinetic characteristics (Michaelis constant of aortic 15-LO is 2.2±0.3 μmol/L for AA and 23.5±3.3 μmol/L for linoleic acid) of aortic 15-LO were similar to those of the purified 15-LO-I. An antisense oligonucleotide inhibited 15-LO-I expression in rabbit aorta. Indomethacin and nitro-l-arginine-resistant relaxations to acetylcholine were inhibited by 15-LO-I antisense oligonucleotide but not by the scrambled oligonucleotide.
CONCLUSIONS—15-LO-I is expressed in rabbit aortic endothelium and is important in endothelium-dependent regulation of vascular tone.</description><identifier>ISSN: 1079-5642</identifier><identifier>EISSN: 1524-4636</identifier><identifier>DOI: 10.1161/01.ATV.0000191640.73313.ad</identifier><identifier>PMID: 16239596</identifier><identifier>CODEN: ATVBFA</identifier><language>eng</language><publisher>Philadelphia, PA: American Heart Association, Inc</publisher><subject>Acetylcholine - pharmacology ; Amino Acid Sequence ; Animals ; Aorta - cytology ; Aorta - enzymology ; Arachidonate 12-Lipoxygenase - genetics ; Arachidonate 15-Lipoxygenase - genetics ; Arachidonate 15-Lipoxygenase - metabolism ; Atherosclerosis (general aspects, experimental research) ; Biological and medical sciences ; Blood and lymphatic vessels ; Cardiology. Vascular system ; Cells, Cultured ; Cytosol - enzymology ; Diseases caused by cestodes ; Diseases of the peripheral vessels. Diseases of the vena cava. Miscellaneous ; Drug toxicity and drugs side effects treatment ; Echinococcoses ; Endothelium, Vascular - cytology ; Endothelium, Vascular - enzymology ; Gene Expression Regulation, Enzymologic - physiology ; Helminthic diseases ; Humans ; Infectious diseases ; Isometric Contraction - physiology ; Medical sciences ; Molecular Sequence Data ; Muscle, Smooth, Vascular - cytology ; Muscle, Smooth, Vascular - physiology ; Parasitic diseases ; Pharmacology. Drug treatments ; Rabbits ; Reticulocytes - enzymology ; Toxicity: blood ; Vasodilation - drug effects ; Vasodilation - physiology ; Vasodilator Agents - pharmacology</subject><ispartof>Arteriosclerosis, thrombosis, and vascular biology, 2006-01, Vol.26 (1), p.78-84</ispartof><rights>2006 American Heart Association, Inc.</rights><rights>2006 INIST-CNRS</rights><rights>Copyright American Heart Association, Inc. Jan 2006</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4514-aec7a839eeaa2b8a28ee22e1ddfaaf71cfb275111415de5c08d4424c06bbd9a03</citedby><cites>FETCH-LOGICAL-c4514-aec7a839eeaa2b8a28ee22e1ddfaaf71cfb275111415de5c08d4424c06bbd9a03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,4009,27902,27903,27904</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=17441892$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16239596$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Tang, Xin</creatorcontrib><creatorcontrib>Holmes, Blythe B</creatorcontrib><creatorcontrib>Nithipatikom, Kasem</creatorcontrib><creatorcontrib>Hillard, Cecilia J</creatorcontrib><creatorcontrib>Kuhn, Hartmut</creatorcontrib><creatorcontrib>Campbell, William B</creatorcontrib><title>Reticulocyte 15-Lipoxygenase-I Is Important in Acetylcholine-Induced Endothelium-Dependent Vasorelaxation in Rabbit Aorta</title><title>Arteriosclerosis, thrombosis, and vascular biology</title><addtitle>Arterioscler Thromb Vasc Biol</addtitle><description>OBJECTIVE—Aortic 15-lipoxygenase (15-LO) metabolizes arachidonic acid (AA) to 15-hydroperoxyeicosatetraenoic acid, which is then converted to the vasodilators 15-hydroxy-11,12-epoxyeicosatrienoic acid and 11,12,15-trihydroxyeicosatrienoic acid. These metabolites contribute to endothelium-dependent relaxations of rabbit aorta to AA and acetylcholine. We investigated the identity of rabbit aortic 15-LO and studied its importance in the regulation of vascular tone.
METHODS AND RESULTS—RT-PCR using 12-lipoxygenase/15-LO specific primers resulted in a 572-bp product with a sequence identical to 15-LO-I from rabbit aorta. A RT-PCR/restriction digest strategy excluded expression of 12-lipoxygenase. Immunoblotting revealed 15-LO-I expression in rabbit endothelial and smooth muscle cells. Aortic homogenates and cytosolic fractions metabolize AA to 15(S)-hydroxyeicosatetraenoic acid and linoleic acid to 13(S)-hydroxyoctadecadienoic acid. This activity was blocked by LO inhibitors. The kinetic characteristics (Michaelis constant of aortic 15-LO is 2.2±0.3 μmol/L for AA and 23.5±3.3 μmol/L for linoleic acid) of aortic 15-LO were similar to those of the purified 15-LO-I. An antisense oligonucleotide inhibited 15-LO-I expression in rabbit aorta. Indomethacin and nitro-l-arginine-resistant relaxations to acetylcholine were inhibited by 15-LO-I antisense oligonucleotide but not by the scrambled oligonucleotide.
CONCLUSIONS—15-LO-I is expressed in rabbit aortic endothelium and is important in endothelium-dependent regulation of vascular tone.</description><subject>Acetylcholine - pharmacology</subject><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Aorta - cytology</subject><subject>Aorta - enzymology</subject><subject>Arachidonate 12-Lipoxygenase - genetics</subject><subject>Arachidonate 15-Lipoxygenase - genetics</subject><subject>Arachidonate 15-Lipoxygenase - metabolism</subject><subject>Atherosclerosis (general aspects, experimental research)</subject><subject>Biological and medical sciences</subject><subject>Blood and lymphatic vessels</subject><subject>Cardiology. Vascular system</subject><subject>Cells, Cultured</subject><subject>Cytosol - enzymology</subject><subject>Diseases caused by cestodes</subject><subject>Diseases of the peripheral vessels. Diseases of the vena cava. Miscellaneous</subject><subject>Drug toxicity and drugs side effects treatment</subject><subject>Echinococcoses</subject><subject>Endothelium, Vascular - cytology</subject><subject>Endothelium, Vascular - enzymology</subject><subject>Gene Expression Regulation, Enzymologic - physiology</subject><subject>Helminthic diseases</subject><subject>Humans</subject><subject>Infectious diseases</subject><subject>Isometric Contraction - physiology</subject><subject>Medical sciences</subject><subject>Molecular Sequence Data</subject><subject>Muscle, Smooth, Vascular - cytology</subject><subject>Muscle, Smooth, Vascular - physiology</subject><subject>Parasitic diseases</subject><subject>Pharmacology. Drug treatments</subject><subject>Rabbits</subject><subject>Reticulocytes - enzymology</subject><subject>Toxicity: blood</subject><subject>Vasodilation - drug effects</subject><subject>Vasodilation - physiology</subject><subject>Vasodilator Agents - pharmacology</subject><issn>1079-5642</issn><issn>1524-4636</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdkdGK1DAUhoso7rr6ClIW9K5jkiZp692wrjowICzr3obT5NTpmknGJGW3b2_qDAyYmyTk-8855CuKa0pWlEr6idDV-v5hRfKiHZWcrJq6pvUKzIvikgrGKy5r-TKfSdNVQnJ2UbyJ8THznDHyurigktWd6ORlMd9hGvVkvZ4TllRU2_Hgn-df6CBitSk3sdzsDz4kcKkcXbnWmGard96OLr87M2k05a0zPu3QjtO--oIHdAYz_gDRB7TwDGn0bknfQd-PqVwv9d4WrwawEd-d9qvi59fb-5vv1fbHt83NeltpLiivAHUDbd0hArC-BdYiMobUmAFgaKgeetYISimnwqDQpDWcM66J7HvTAamvio_Huofg_0wYk9qPUaO14NBPUclGtA0TIoPX_4GPfgouz6ZY_ri2lfUCfT5COvgYAw7qEMY9hFlRohY7ilCV7aizHfXPjgKTw-9PHaZ-j-YcPenIwIcTAFGDHQI4PcYz13BO245ljh-5J28ThvjbTk8Y1A7Bpt3SmteSiIoRIgnN12oZhtd_AT-eqjA</recordid><startdate>200601</startdate><enddate>200601</enddate><creator>Tang, Xin</creator><creator>Holmes, Blythe B</creator><creator>Nithipatikom, Kasem</creator><creator>Hillard, Cecilia J</creator><creator>Kuhn, Hartmut</creator><creator>Campbell, William B</creator><general>American Heart Association, Inc</general><general>Lippincott</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>K9.</scope><scope>7X8</scope></search><sort><creationdate>200601</creationdate><title>Reticulocyte 15-Lipoxygenase-I Is Important in Acetylcholine-Induced Endothelium-Dependent Vasorelaxation in Rabbit Aorta</title><author>Tang, Xin ; Holmes, Blythe B ; Nithipatikom, Kasem ; Hillard, Cecilia J ; Kuhn, Hartmut ; Campbell, William B</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4514-aec7a839eeaa2b8a28ee22e1ddfaaf71cfb275111415de5c08d4424c06bbd9a03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Acetylcholine - pharmacology</topic><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Aorta - cytology</topic><topic>Aorta - enzymology</topic><topic>Arachidonate 12-Lipoxygenase - genetics</topic><topic>Arachidonate 15-Lipoxygenase - genetics</topic><topic>Arachidonate 15-Lipoxygenase - metabolism</topic><topic>Atherosclerosis (general aspects, experimental research)</topic><topic>Biological and medical sciences</topic><topic>Blood and lymphatic vessels</topic><topic>Cardiology. Vascular system</topic><topic>Cells, Cultured</topic><topic>Cytosol - enzymology</topic><topic>Diseases caused by cestodes</topic><topic>Diseases of the peripheral vessels. Diseases of the vena cava. Miscellaneous</topic><topic>Drug toxicity and drugs side effects treatment</topic><topic>Echinococcoses</topic><topic>Endothelium, Vascular - cytology</topic><topic>Endothelium, Vascular - enzymology</topic><topic>Gene Expression Regulation, Enzymologic - physiology</topic><topic>Helminthic diseases</topic><topic>Humans</topic><topic>Infectious diseases</topic><topic>Isometric Contraction - physiology</topic><topic>Medical sciences</topic><topic>Molecular Sequence Data</topic><topic>Muscle, Smooth, Vascular - cytology</topic><topic>Muscle, Smooth, Vascular - physiology</topic><topic>Parasitic diseases</topic><topic>Pharmacology. Drug treatments</topic><topic>Rabbits</topic><topic>Reticulocytes - enzymology</topic><topic>Toxicity: blood</topic><topic>Vasodilation - drug effects</topic><topic>Vasodilation - physiology</topic><topic>Vasodilator Agents - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Tang, Xin</creatorcontrib><creatorcontrib>Holmes, Blythe B</creatorcontrib><creatorcontrib>Nithipatikom, Kasem</creatorcontrib><creatorcontrib>Hillard, Cecilia J</creatorcontrib><creatorcontrib>Kuhn, Hartmut</creatorcontrib><creatorcontrib>Campbell, William B</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>MEDLINE - Academic</collection><jtitle>Arteriosclerosis, thrombosis, and vascular biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Tang, Xin</au><au>Holmes, Blythe B</au><au>Nithipatikom, Kasem</au><au>Hillard, Cecilia J</au><au>Kuhn, Hartmut</au><au>Campbell, William B</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Reticulocyte 15-Lipoxygenase-I Is Important in Acetylcholine-Induced Endothelium-Dependent Vasorelaxation in Rabbit Aorta</atitle><jtitle>Arteriosclerosis, thrombosis, and vascular biology</jtitle><addtitle>Arterioscler Thromb Vasc Biol</addtitle><date>2006-01</date><risdate>2006</risdate><volume>26</volume><issue>1</issue><spage>78</spage><epage>84</epage><pages>78-84</pages><issn>1079-5642</issn><eissn>1524-4636</eissn><coden>ATVBFA</coden><abstract>OBJECTIVE—Aortic 15-lipoxygenase (15-LO) metabolizes arachidonic acid (AA) to 15-hydroperoxyeicosatetraenoic acid, which is then converted to the vasodilators 15-hydroxy-11,12-epoxyeicosatrienoic acid and 11,12,15-trihydroxyeicosatrienoic acid. These metabolites contribute to endothelium-dependent relaxations of rabbit aorta to AA and acetylcholine. We investigated the identity of rabbit aortic 15-LO and studied its importance in the regulation of vascular tone.
METHODS AND RESULTS—RT-PCR using 12-lipoxygenase/15-LO specific primers resulted in a 572-bp product with a sequence identical to 15-LO-I from rabbit aorta. A RT-PCR/restriction digest strategy excluded expression of 12-lipoxygenase. Immunoblotting revealed 15-LO-I expression in rabbit endothelial and smooth muscle cells. Aortic homogenates and cytosolic fractions metabolize AA to 15(S)-hydroxyeicosatetraenoic acid and linoleic acid to 13(S)-hydroxyoctadecadienoic acid. This activity was blocked by LO inhibitors. The kinetic characteristics (Michaelis constant of aortic 15-LO is 2.2±0.3 μmol/L for AA and 23.5±3.3 μmol/L for linoleic acid) of aortic 15-LO were similar to those of the purified 15-LO-I. An antisense oligonucleotide inhibited 15-LO-I expression in rabbit aorta. Indomethacin and nitro-l-arginine-resistant relaxations to acetylcholine were inhibited by 15-LO-I antisense oligonucleotide but not by the scrambled oligonucleotide.
CONCLUSIONS—15-LO-I is expressed in rabbit aortic endothelium and is important in endothelium-dependent regulation of vascular tone.</abstract><cop>Philadelphia, PA</cop><cop>Hagerstown, MD</cop><pub>American Heart Association, Inc</pub><pmid>16239596</pmid><doi>10.1161/01.ATV.0000191640.73313.ad</doi><tpages>7</tpages></addata></record> |
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| ispartof | Arteriosclerosis, thrombosis, and vascular biology, 2006-01, Vol.26 (1), p.78-84 |
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| subjects | Acetylcholine - pharmacology Amino Acid Sequence Animals Aorta - cytology Aorta - enzymology Arachidonate 12-Lipoxygenase - genetics Arachidonate 15-Lipoxygenase - genetics Arachidonate 15-Lipoxygenase - metabolism Atherosclerosis (general aspects, experimental research) Biological and medical sciences Blood and lymphatic vessels Cardiology. Vascular system Cells, Cultured Cytosol - enzymology Diseases caused by cestodes Diseases of the peripheral vessels. Diseases of the vena cava. Miscellaneous Drug toxicity and drugs side effects treatment Echinococcoses Endothelium, Vascular - cytology Endothelium, Vascular - enzymology Gene Expression Regulation, Enzymologic - physiology Helminthic diseases Humans Infectious diseases Isometric Contraction - physiology Medical sciences Molecular Sequence Data Muscle, Smooth, Vascular - cytology Muscle, Smooth, Vascular - physiology Parasitic diseases Pharmacology. Drug treatments Rabbits Reticulocytes - enzymology Toxicity: blood Vasodilation - drug effects Vasodilation - physiology Vasodilator Agents - pharmacology |
| title | Reticulocyte 15-Lipoxygenase-I Is Important in Acetylcholine-Induced Endothelium-Dependent Vasorelaxation in Rabbit Aorta |
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