title: MDA, oxypurines, and nucleosides relate to reperfusion in short-term incomplete cerebral ischemia in the rat.
abstract: Short-term incomplete cerebral ischemia (5 min) was induced in the rat by the bilateral clamping of the common carotid arteries. Reperfusion was obtained by removing carotid clamping and was carried out for the following 10 min. Animals were sacrificed either at the end of ischemia or reperfusion. Controls were represented by a group of sham-operated rats. Peripheral venous blood samples were withdrawn from the femoral vein from rats subjected to cerebral reperfusion 5 min before ischemia, at the end of ischemia, and 10 min after reperfusion. Neutralized perchloric acid extracts of brain tissue were analyzed by a highly sensitive high-performance liquid chromatography (HPLC) method for the direct determination of malondialdehyde, oxypurines, nucleosides, nicotinic coenzymes, and high-energy phosphates. In addition, plasma concentrations of malondialdehyde, hypoxanthine, xanthine, inosine, uric acid, and adenosine were determined by the same HPLC technique. Incomplete cerebral ischemia induced the appearance of a significant amount (8.05 nmol/g w.w.; SD = 2.82) of cerebral malondialdehyde (which was undetectable in control animals) and a decrease of ascorbic acid. A further 6.6-fold increase of malondialdehyde (53.30 nmol/g w.w.; SD = 17.77) and a 18.5% decrease of ascorbic acid occurred after 10 min of reperfusion. Plasma malondialdehyde, which was present in minimal amount before ischemia (0.050 mumol/L; SD = 0.015), significantly increased after 5 min of ischemia (0.277 mumol/L; SD = 0.056) and was strikingly augmented after 10 min of reperfusion (0.682 mumol/L; SD = 0.094). A similar trend was observed for xanthine, uric acid, inosine, and adenosine, while hypoxanthine reached its maximal concentration after 5 min of incomplete ischemia, being significantly decreased after reperfusion. From the data obtained, it can be concluded that tissue concentrations of malondialdehyde and ascorbic acid, and plasma levels of malondialdehyde, oxypurines, and nucleosides, reflect both the oxygen radical-mediated tissue injury and the depression of energy metabolism, thus representing early biochemical markers of short-term incomplete brain ischemia and reperfusion in the rat. In particular, these results suggest the possibility of using the variation of malondialdehyde, oxypurines, and nucleosides in peripheral blood as a potential biochemical indicator of reperfusion damage occurring to postischemic tissues.
authors.affiliation: Department of Experimental Medicine and Biochemical Sciences, University of Rome Tor Vergata, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Lazzarino
authors.forename: R
authors.initials: R
authors.lastname: Vagnozzi
authors.forename: B
authors.initials: B
authors.lastname: Tavazzi
authors.forename: F S
authors.initials: FS
authors.lastname: Pastore
authors.forename: D
authors.initials: D
authors.lastname: Di Pierro
authors.forename: P
authors.initials: P
authors.lastname: Siragusa
authors.forename: A
authors.initials: A
authors.lastname: Belli
authors.forename: R
authors.initials: R
authors.lastname: Giuffré
authors.forename: B
authors.initials: B
authors.lastname: Giardina
mesh_terms: D000227:Adenine Nucleotides / Q000097:blood / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000378:metabolism*; D002339:Carotid Arteries / Q000502:physiology; D002851:Chromatography, High Pressure Liquid; D005609:Free Radicals; D002546:Ischemic Attack, Transient / Q000097:blood / Q000378:metabolism*; D008297:Male; D008315:Malondialdehyde / Q000097:blood / Q000378:metabolism*; D009705:Nucleosides / Q000097:blood / Q000378:metabolism*; D011685:Purine Nucleotides / Q000097:blood / Q000378:metabolism*; D011687:Purines / Q000097:blood / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D012016:Reference Values; D015424:Reperfusion; D013997:Time Factors
chemical_list: D000227:Adenine Nucleotides; D005609:Free Radicals; D009705:Nucleosides; D011685:Purine Nucleotides; D011687:Purines; D008315:Malondialdehyde