Nitric Oxide and Lipid Peroxidation

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1 Nitric Oxide and Lipid Peroxidation Valerie B. O Donnell a, Neil Hogg b and Victor M. Darley Usmar c a Dept of Med. Biochem. University of Wales College of Medicine, Heath Park, Cardiff CF14 4XN, UK. Ph: Fax: o-donnellvb@cardiff.ac.uk b Biophysics Research Institute, Medical College of Wisconsin, Milwaukee, WI USA. c Department of Pathology, University of Alabama at Birmingham, AL USA NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 1

2 Nitric Oxide is a Free Radical N=O NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 2

3 Nitric oxide is not particularly oxidizing. Nitric oxide is (infinitely) stable*. Nitric oxide does not rapidly react with organic molecules. *In the absence of dioxygen. NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 3

4 The Nitric Oxide Radical and some of its stable end-products N=O nitric oxide - O-N=O nitrite RO-N=O alkyl nitrite RS-N=O nitrosothiol NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 4

5 Reactions of NO with Other Free Radicals Oxidation of NO to nitrite: 4 NO+ O=O +2H 2 O 4HNO 2 Nitric oxide reaction with organic peroxyl radicals: NO+ ROO ROONO?? NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 5

6 Lipid Peroxidation LOO + LH + O 2 LOO + LOOH LOO is the central and rate-limiting species of lipid peroxidation NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 6

7 LOOH Non-enzymatic lipid oxidation Metal or enzyme AOO O 2 LOO LH L LOOH O 2 LOO LH L LOOH O 2 LOO LH GPx GPx Initiation LOH Propagation LOH Propagation NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 7

8 Lipid peroxidation products in an atherosclerotic lesion are both enzymatically and nonenzymatically produced. Foam cell 15-LOX Necrotic core lipid hydroxides, hydroperoxides, aldehydes, epoxides, ketones, ceroid, isoprostanes Fibrous cap NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 8

9 Location of lipid oxidation enzymes in the vasculature LOX: lipoxygenase PGHS: prostaglandin H synthase Smooth muscle: 12-LOX Endothelium PGHS-1 Monocyte: PGHS-1,2 5, 15-LOX Platelet: PGHS-1 12-LOX Neutrophil: PGHS-1,2 5,12-LOX NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 9

10 Lipid oxidation enzymes are upregulated during vascular disease. * Inhibition of 12-LOX restores blood pressure in Angiotensin- II dependent hypertensive rats. * Increased products of PGHS in hypertensive rats, and PGHS inhibition restores blood pressure in humans and rats. * Smooth muscle 12-LOX in hypertension * Platelet 12-LOX in hypertension * Foam cell/macrophage 15-LOX in atherosclerosis. NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 10

11 Nitric oxide reaction with lipid peroxyl radicals LOO NO LOONO H 2 O [LO NO 2 ] LOOH + NO 2 - LO NO 2 NO [L(O) NO 2 ] NO LONO NO - 2 L(O)NO 2 NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 11

12 Reactions between NO-derived reactive nitrogen species and unsaturated lipid NO LOONO L(O)NO 2 O 2 O 2 - NO 2 ONOO - NO HONO LH O L 2 LOO e -, H + LOOH NO 2, ONOO - LNO 2 NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 12

13 Nitric oxide inhibition of lipid oxidation coincides with fast rates of NO uptake during ABAP-dependent linoleate oxidation. (A) (B) Oxygen (µm) 100 ABAP 0.2 NO (µm) [ NO] = 0.9 µm added at vertical lines to 7.3 mm linoleate in 0.5% cholate, 11 mm ABAP Time/s NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 13

14 Mechanism of Inhibition of LDL Oxidation by NO LOO + NO LOONO LO + NO 2 LONO 2 LOOH OLOO + NO OLOONO OLONO 2 Two NO consumed for each LOO 4 NO + 2LOO + H 2 O 2HNO 2 + LONO 2 + LONO NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 14

15 Cholesteryl Ester Triglyceride Phospholipid Apolipoprotein B Oxidant LH NO 2 O 2 O 2 L NO 2 LH LOO NO LNO 2 2 L α-tocopherol LOOH NO L(O)NO 2 Cholesterol NO NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 15

16 500 NO and LDL Oxidation: The effect of HPODE Lag time of LDL (125 µg/ml) oxidation by Cu(II) (20 µm) with and without HPODE (5 µm) length of lag time (min) A SNN SNN + HpODE SNN (µm) B BHT BHT + HpODE BHT (µm) NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 16

17 Comparison of nitric oxide and tocopherol inhibition of linoleate oxidation. 300 ABAP α -tocopherol (1 µm) [Oxygen]/µM T inh NO (1 µm) T inh Time/s Oxidation of linoleate (7.3 mm) by ABAP (11 mm) NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 17

18 Nitric oxide is consumed by rabbit 15-lipoxygenase during oxidation of linoleate. 2 LOX LOX LOX NO (µm) 1 Add figure caption 0 (i) (ii) (iii) Time (secs) NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 18

19 Site of lipoxygenase inhibition by NO E-Fe 2+ LOOH E-Fe 3+ NO E-Fe 3+ LOO E-Fe 2+ L O 2 E-Fe 2+ LOO NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 19

20 Activation of LOX inhibits activation of sgc by NO NO Lox 100 sgc cgmp cgmp (% control) 50 ** 0 control AA, 5 µm LOX AA, 5 µm, LOX NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 20

21 Further Reading: Darley-Usmar, VM et al., (1992) Free Radic. Res. Commun. 17, 9-20 Graham, A et al., (1993) FEBS Lett. 330, Rubbo, H et al., (1994) J. Biol. Chem. 265, Hogg, N et al., (1995) J. Lipid Res. 36, Struck, N et al., (1995) FEBS Lett 361, O Donnell, VB et al., (1997) Biochemistry 36, O Donnell, VB et al., (1999) Chem. Res. Toxicol. 12, O Donnell, VB et al., (1999) J. Biol. Chem. 274, Hogg, N & Kalyanaraman, B (1999) Biochim. Biophys. Acta. 1411, Rubbo, H et al., (2000) J. Biol. Chem. 275, O Donnell, VB & Freeman, BA (2001) Circ. Res. 88, NO & LPO Society For Free Radical Biology and Medicine O Donnell et al. 21

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