{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T11:34:58Z","timestamp":1791286498244,"version":"4.1.0"},"reference-count":73,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2004,7,15]],"date-time":"2004-07-15T00:00:00Z","timestamp":1089849600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Journal of Neurochemistry"],"published-print":{"date-parts":[[2004,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Cyclo\u2010oxygenase\u20102 (COX\u20102) is believed to induce neuronal oxidative stress via production of radicals. While oxygen radicals are not directly involved in COX\u20102\u2010catalytic cycle, superoxide anion radicals have been repeatedly reported to play a critical role in COX\u20102\u2010associated oxidative stress. To resolve the controversy, we characterized production of free radicals in PC12 cells in which COX\u20102 expression was manipulated either genetically or by direct protein transfection and compared them with those generated by a recombinant COX\u20102 in a cell\u2010free system. Using spin\u2010traps \u03b1\u2010(4\u2010pyridyl\u20101\u2010oxide)\u2010<jats:italic>N<\/jats:italic>\u2010t\u2010butylnitrone, 5,5\u2010dimethyl\u20101\u2010pyrroline\u2010<jats:italic>N<\/jats:italic>\u2010oxide and 4\u2010((9\u2010acridinecarbonyl) amino)\u20102,2,6,6\u2010 tetramethylpiperidine\u20101\u2010oxyl (Ac\u2010Tempo), we observed arachidonic acid (AA)\u2010dependent production of carbon\u2010centered radicals by heme\u2010reconstituted recombinant COX\u20102. No oxygen radicals or thiyl radicals have been detected. COX\u20102 also catalyzed AA\u2010dependent one\u2010electron co\u2010oxidation of ascorbate to ascorbate radicals. Next, we used two different approaches of COX\u20102 expression in cells, PCXII cells which express isopropyl\u20101\u2010thio\u2010\u03b2\u2010<jats:sc>d<\/jats:sc>\u2010galactopyranoside inducible COX\u20102, and PC12 cells transfected with COX\u20102 using a protein delivery reagent, Chariot. In both models, COX\u20102\u2010dependent AA\u2010induced generation of carbon\u2010centered radicals was documented using spin\u2010traps and Ac\u2010Tempo. No oxygen radical formation was detected in COX\u20102\u2010transfected cells by either spin\u2010traps or fluorogenic probe, dihydroethidium. In the presence of ascorbate, AA\u2010induced COX\u20102\u2010dependent ascorbate radicals were detected. AA caused a significant and selective oxidation of one of the major phospholipids, phosphatidylserine (PS). PS was not a direct substrate for COX\u20102 but was co\u2010oxidized in the presence of AA. The radical generation and PS oxidation were inhibited by COX\u20102 inhibitors, niflumic acid, nimesulide, or NS\u2010398. Thus, COX\u20102 generated carbon\u2010centered radicals but not oxygen radicals or thiyl radicals are responsible for oxidative stress in AA\u2010challenged PC12 cells overexpressing COX\u20102.<\/jats:p>","DOI":"10.1111\/j.1471-4159.2004.02577.x","type":"journal-article","created":{"date-parts":[[2004,8,16]],"date-time":"2004-08-16T14:44:00Z","timestamp":1092667440000},"page":"1036-1049","source":"Crossref","is-referenced-by-count":55,"title":["Arachidonic acid\u2010induced carbon\u2010centered radicals and phospholipid peroxidation in cyclo\u2010oxygenase\u20102\u2010transfected PC12 cells"],"prefix":"10.1111","volume":"90","author":[{"given":"Jianfei","family":"Jiang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Grigory G.","family":"Borisenko","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anatoly","family":"Osipov","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ian","family":"Martin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renwu","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna A.","family":"Shvedova","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrey","family":"Sorokin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yulia Y.","family":"Tyurina","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alla","family":"Potapovich","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vladimir A.","family":"Tyurin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven H.","family":"Graham","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valerian E.","family":"Kagan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2004,7,27]]},"reference":[{"key":"e_1_2_41_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-8993(02)02447-2"},{"key":"e_1_2_41_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1054-3589(08)60274-4"},{"key":"e_1_2_41_4_1","doi-asserted-by":"publisher","DOI":"10.1097\/00000542-200306000-00012"},{"key":"e_1_2_41_5_1","first-page":"H401","article-title":"Postischemic generation of superoxide anion by newborn pig brain","volume":"255","author":"Armstead W. 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