Peroxynitrite-induced membrane lipid peroxidation: the cytotoxic potential of superoxide and nitric oxide.

Radi R, Beckman JS, Bush KM, Freeman BA. Source Department of Anesthesiology, A"stract &ndothelial cells, macrophages, neutrophils, and neuronal cells generate supero'ide ()%*+ and nitric o'ide (.,)+ -hich can com"ine to form pero'ynitrite anion (),))*+. .ero'ynitrite, kno-n to o'idi/e sulfhydryls and to yield products indicati!e of hydro'yl radical (.)0+ reaction -ith deo'yri"ose and dimethyl sulfo'ide, is sho-n herein to induce mem"rane lipid pero'idation. .ero'ynitrite addition to soy"ean phosphatidylcholine liposomes resulted in malondialdehyde and con1ugated diene formation, as -ell as o'ygen consumption. 2ipid pero'idation -as greater at acidic and neutral p0, -ith no significant lipid pero'idation occurring a"o!e p0 3.$. Addition of ferrous (Fe4%+ or ferric (Fe4#+ iron did not enhance lipid pero'ide formation o!er that attri"uta"le to pero'ynitrite alone. Diethylenetetraminepentacetic acid (D5.A+ or iron remo!al from solutions "y ion* e'change chromatography decreased con1ugated diene formation "y %$*$67. 8ron did not play an essential role in initiating lipid pero'idation, since D5.A and iron depletion of reaction systems -ere only partially inhi"itory. 8n contrast, desferrio'amine had an e!en greater concentration*dependent inhi"itory effect, completely a"olishing lipid pero'idation at %66 microM. 5he strong inhi"itory effect of desferrio'amine on lipid pero'idation -as due to direct reaction -ith pero'ynitrous acid in addition to iron chelation. 9e conclude that the con1ugate acid of pero'ynitrite, pero'ynitrous acid (),))0+, and:or its decomposition products, i.e., .)0 and nitrogen dio'ide (.,)%+, initiate lipid pero'idation -ithout the re;uirement of iron. 5hese o"ser!ations demonstrate a potential mechanism contri"uting to )%*(*+and .,)*mediated cytoto'icity. ni!ersity of Ala"ama, Birmingham #$%##.

Peroxynitrite-mediated protein nitration and lipid peroxidation in a mouse model of traumatic brain injur y .
0all &D, Detloff MR, Johnson K, Kupina ,<. Source Spinal <ord and Brain 8n1ury Research <enter, ni!ersity of Kentucky <handler Medical <enter, 2e'ington, Kentucky =6$#>*6#6$, SA. edhall?uky.edu A"stract

5he role of reacti!e o'ygen*induced o'idati!e damage to lipids (i.e., lipid pero'idation, 2.+ and proteins has "een strongly supported in pre!ious -ork. Most nota"ly, a num"er of free radical sca!engers and lipid antio'idants ha!e "een demonstrated to "e neuroprotecti!e in traumatic "rain in1ury (5B8+ models. 0o-e!er, the specific sources of reacti!e o'ygen species (R)S+, the time course of o'idati!e damage and its relationship to post*traumatic neurodegeneration in the in1ured "rain ha!e "een incompletely defined. 5he present study -as directed at an in!estigation of the role of the R)S, pero'ynitrite (.),+, in the acute pathophysiology of 5B8 and its temporal relationship to neurodegeneration in the conte't of the mouse model of diffuse head in1ury model. Male <F*@ mice -ere su"1ected to a moderately se!ere head in1ury and assessed at @*, #*, >*, @%*, %=*, =A*, B%, 3>* and @%6*h post*in1ury for neurodegeneration using ;uantitati!e image analysis of sil!er staining and semi*;uantitati!e analysis of .),*mediated o'idati!e damage to proteins (#*nitrotyrosine, #*,5+ and lipids (=*hydro'ynonenal, =* 0,&+. Significant e!idence of sil!er staining -as not apparent until %=*h post*in1ury, -ith peak staining seen "et-een B%* and @%6*h. 5his time*course of neurodegeneration -as preceded "y intense immunostaining for #*,5 and =*0,&, -hich occurred -ithin the first hour post*in1ury. 5he time course and staining pattern for #*,5 and =*0,& -ere similar, -ith the highest staining intensity noted -ithin the first =A*h in areas surrounding trauma* induced contusions. 8n the case of #*,5, neuronal perikarya and processes and micro!essels displayed staining. 5he temporal and spatial coincidence of protein nitration and 2. damage suggests that .), is in!ol!ed in "oth. 0o-e!er, lipid* pero'idati!e (=*0,&+ immunoreacti!ity -as "roader and more diffuse than #*,5, suggesting that other reacti!e o'ygen mechanisms, such as iron*dependent 2., may also contri"ute to the more -idespread =*0,& immunoreacti!ity. 5his indicates that optimal pharmacological inhi"ition of post*traumatic o'idati!e damage in 5B8 may need to com"ine t-o functionalitiesC one to sca!enge .), or .),*deri!ed radicals, and the second to inhi"it 2. caused "y multiple R)S species.

Peroxynitrite induces contractile dysfunction and lipid peroxidation in the diaphragm . Supinski D, Stofan D, <allahan 2A, ,ethery D, ,osek 5M, DiMarco A. Source .ulmonary Di!ision, Department of Medicine, <ase 9estern Reser!e ni!ersity, <le!eland ==@6>E and Metrohealth Medical <enter, <le!eland, )hio ==@63, SA. A"stract .ero'ynitrite may "e generated in and around muscles in se!eral pathophysiological conditions (e.g., sepsis+ and may induce muscle dysfunction in these disease states. 5he effect of pero'ynitrite on muscle force generation has not "een directly assessed. 5he purpose of the present study -as to assess the effects of pero'ynitrite

administration on diaphragmatic force*generating capacity in @+ intact diaphragm muscle fi"er "undles (to model the effects produced "y e'posure of muscles to e'tracellular pero'ynitrite+ and %+ single skinned diaphragm muscle fi"ers (to model the effects of intracellular pero'ynitrite on contractile protein function+ "y e'amining the effects of "oth pero'ynitrite and a pero'ynitrite*generating solution, #*morpholinosydnonimine, on force !s. p<a characteristics. 8n intact diaphragm preparations, pero'ynitrite reduced diaphragm force generation and increased muscle le!els of =*hydro'ynonenal (an inde' of lipid pero'idation+. 8n skinned fi"ers, "oth pero'ynitrite and #*morpholinosydnonimine reduced ma'imum calcium*acti!ated force. 5hese data indicate that pero'ynitrite is capa"le of producing significant diaphragmatic contractile dysfunction. 9e speculate that pero'ynitrite*mediated alterations may "e responsi"le for much of the muscle dysfunction seen in pathophysiological conditions such as sepsis.

Nitric oxide and peroxynitrite in lipid peroxidation . Ru""o 0. Source Departamento de Bio;uFmica, Facultad de Medicina, Monte!ideo, ruguay. hru""o?fmed.edu.uy A"stract ,itric o'ide (.,)+ can mediate tissue protecti!e reactions during o'idant stress, as -ell as to'ic and tissue proo'idant effects. ,itric o'ide regulates critical lipid mem"rane and lipoprotein o'idation e!ents, "y @+ contri"uting to the formation of more potent secondary o'idants from supero'ide (i.e. pero'ynitrite+ and %+ termination of lipid radicals to possi"ly less reacti!e secondary nitrogen*containing products (2),), 2)),)+ -hich are in part organic pero'ynitrites and are e'pected to "e produced in !i!o. Relati!e rates of production and steady state concentrations of supero'ide and .,) and cellular sites of production -ill profoundly influence e'pression of the differential o'idant in1ury* enhancing and protecti!e effects of .,). Full understanding of the physiological roles of .,), coupled -ith detailed insight into .,) regulation of o'ygen radical*dependent reactions, -ill yield a more rational "asis for the use of .,) donors for therapeutic purposes. ni!ersidad de la RepG"lica,

Chapter One The A"stract

iological Chemistr y of !ydrogen Peroxide

0ydrogen pero'ide is generated in numerous "iological processes and is implicated as the main transmitter of redo' signals. Although a strong o'idant, high acti!ation energy "arriers make it unreacti!e -ith most "iological molecules. 8t reacts directly -ith thiols, "ut for lo-*molecular*-eight thiols and cysteine residues in most proteins, the reaction is slo-. 5he most fa!ored reactions of hydrogen pero'ide are -ith transition metal centers, selenoproteins, and selected thiol proteins. 5hese include proteins such as catalase, glutathione pero'idases, and pero'iredo'ins, -hich, as -ell as pro!iding antio'idant defense, are increasingly "eing considered as targets for signal transmission. 5his o!er!ie- descri"es the main "iological reactions of hydrogen pero'ide and takes a kinetic approach to identifying likely targets in the cell. 8t also considers diffusion of hydrogen pero'ide and constraints to its acting at locali/ed sites.