Oxidized LDL stimulates lipid peroxidation-derived DNA and protein adducts in human vascular endothelial and smooth muscle cells

Abstract

Oxidized low density lipoprotein (oxLDL) can trigger intracellular production of reactive oxygen species and lipid peroxidation (LPO), and is thought to contribute to initiation and progression of atherosclerosis. In order to understand the correlation between oxLDL and macromolecular damage, we measured levels of LPO-derived miscoding etheno-DNA adducts and LPO-modified proteins in cultured human vascular endothelial and smooth muscle cells after incubation with oxLDL for up to 48 h. A semi-quantative analysis method for 1, N⁶-ethenodeoxyadenosine (ɛdA) by immunohistochemistry was applied. After oxLDL stimulation, ɛdA-stained nuclei were significantly increased in both endothelial and smooth muscle cells. Similarly, 4-hydroxy-2-nonenal (4-HNE)-modified proteins, as analyzed by immunohistochemistry and Western blotting, were also 3–5 fold increased. It was concluded LPO-derived etheno-DNA adducts and LPO-modified proteins are strongly induced by oxLDL in human vascular endothelial and smooth muscle cells. This macromolecular damage may contribute to the dysfunction of arterial endothelium and the onset of atherosclerosis.

Keywords

atherosclerosis / oxidized low density lipoprotein / lipid peroxidation / etheno-DNA adducts / endothelial cell / smooth muscle cell

References

[1] SaundersE, OfiliE. Epidemiology of atherothrombotic disease and the effectiveness and risks of antiplatelet therapy: race and ethnicity considerations. Cardiol Rev, 2008, 16(2): 82-88 PMID: 18281910

[2] De FloraS, QuagliaA, BennicelliC, et al. . The epidemiological revolution of the 20th century. FASEB J, 2005, 19(8): 892-897 PMID: 15923399

[3] FarmerJA. Diabetic dyslipidemia and atherosclerosis: evidence from clinical trials. Curr Diab Rep, 2008, 8(1): 71-77 PMID: 18367002

[4] PaoV, LeeGA, GrunfeldC. HIV therapy, metabolic syndrome, and cardiovascular risk. Curr Atheroscler Rep, 2008, 10(1): 61-70 PMCID: 3166347 PMID: 18366987

[5] AliYS, LintonMF, FazioS. Targeting cardiovascular risk in patients with diabetes: management of dyslipidemia. Curr Opin Endocrinol Diabetes Obes, 2008, 15(2): 142-146 PMID: 18316949

[6] NewbyAC. An overview of the vascular response to injury: a tribute to the late Russell Ross. Toxicol Lett, 2000, 112–113: 519-529 PMID: 10720775

[7] BonettiPO, LermanLO, LermanA. Endothelial dysfunction: a marker of atherosclerotic risk. Arterioscler Thromb Vasc Biol, 2003, 23(2): 168-175 PMID: 12588755

[8] DoranAC, MellerN, McNamaraCA. Role of smooth muscle cells in the initiation and early progression of atherosclerosis. Arterioscler Thromb Vasc Biol, 2008, 28(5): 812-819 PMCID: 2734458 PMID: 18276911

[9] GutierrezJ, BallingerSW, Darley-UsmarVM, et al. . Free radicals, mitochondria, and oxidized lipids: the emerging role in signal transduction in vascular cells. Circ Res, 2006, 99(9): 924-932 PMID: 17068300

[10] NilssonJ, GlazerS, CarlssonR. Antibodies against oxidized low-density lipoprotein for the treatment of vulnerable plaques. Curr Opin Investig Drugs, 2006, 7(9): 815-819 PMID: 17002259

[11] TsimikasS, BrilakisES, MillerER, et al. . Oxidized phospholipids, Lp(a) lipoprotein, and coronary artery disease. N Engl J Med, 2005, 353(1): 46-57 PMID: 16000355

[12] MatsuuraE, HughesGR, KhamashtaMA. Oxidation of LDL and its clinical implication. Autoimmun Rev, 2008, 7(7): 558-566 PMID: 18625445

[13] MatsuuraE, KobayashiK, TabuchiM, et al. . Oxidative modification of low-density lipoprotein and immune regulation of atherosclerosis. Prog Lipid Res, 2006, 45(6): 466-486 PMID: 16790279

[14] PennMS, CuiMZ, WinokurAL, et al. . Smooth muscle cell surface tissue factor pathway activation by oxidized low-density lipoprotein requires cellular lipid peroxidation. Blood, 2000, 96(9): 3056-3063 PMID: 11049984

[15] VindisC, Escargueil-BlancI, ElbazM, et al. . Desensitization of platelet-derived growth factor receptor-beta by oxidized lipids in vascular cells and atherosclerotic lesions: prevention by aldehyde scavengers. Circ Res, 2006, 98(6): 785-792 PMID: 16527993

[16] ThumT, BorlakJ. LOX-1 receptor blockade abrogates oxLDL-induced oxidative DNA damage and prevents activation of the transcriptional repressor Oct-1 in human coronary arterial endothelium. J Biol Chem, 2008, 283(28): 19 456-19 464

[17] NairJ, De FloraS, IzzottiA, et al. . Lipid peroxidation-derived etheno-DNA adducts in human atherosclerotic lesions. Mutat Res, 2007, 621(1–2): 95-105 PMID: 17412369

[18] NairJ, BarbinA, VelicI, et al. . Etheno DNA-base adducts from endogenous reactive species. Mutat Res, 1999, 424(1–2): 59-69 PMID: 10064850

[19] FrankA, SeitzHK, BartschH, et al. . Immunohistochemical detection of 1, N6-ethenodeoxyadenosine in nuclei of human liver affected by diseases predisposing to hepato-carcinogenesis. Carcinogenesis, 2004, 25(6): 1027-1031 PMID: 14742317

[20] YangY, NairJ, BarbinA, et al. . Immunohistochemical detection of 1, N(6)-ethenodeoxyadenosine, a promutagenic DNA adduct, in liver of rats exposed to vinyl chloride or an iron overload. Carcinogenesis, 2000, 21(4): 777-781 PMID: 10753215

[21] KrugerK, JochumC, GlusenkampKH, et al. . New modular delivery system for diagnostic and therapeutic pre-targeting using tautomer-specific monoclonal antibody EM-6-47 and 3-substituted adenines. Int J Cancer, 1998, 77(4): 610-619 PMID: 9679766

[22] PennathurS, HeineckeJW. Oxidative stress and endothelial dysfunction in vascular disease. Curr Diab Rep, 2007, 7(4): 257-264 PMID: 17686400

[23] ChisolmGM, SteinbergD. The oxidative modification hypothesis of atherogenesis: an overview. Free Radic Biol Med, 2000, 28(12): 1815-1826 PMID: 10946223

[24] IulianoL. The oxidant stress hypothesis of atherogenesis. Lipids, 2001, 36Suppl: S41-44

[25] BarbinA. Etheno-adduct-forming chemicals: from mutagenicity testing to tumor mutation spectra. Mutat Res, 2000, 462(2–3): 55-69 PMID: 10767618

[26] GodschalkR, CurfsD, BartschH, et al. . Benzo[a]pyrene enhances lipid peroxidation induced DNA damage in aorta of apolipoprotein E knockout mice. Free Radic Res, 2003, 37(12): 1299-305 PMID: 14753754

[27] AndreassiMG. Coronary atherosclerosis and somatic mutations: an overview of the contributive factors for oxidative DNA damage. Mutat Res, 2003, 543(1): 67-86 PMID: 12510018

[28] AndreassiMG, BottoN, ColomboMG, et al. . Genetic instability and atherosclerosis: can somatic mutations account for the development of cardiovascular diseases?. Environ Mol Mutagen, 2000, 35(4): 265-269 PMID: 10861945

[29] EsterbauerH, RamosP. Chemistry and pathophysiology of oxidation of LDL. Rev Physiol Biochem Pharmacol, 1996, 127: 31-64 PMID: 8533011

[30] BartschH, NairJ. Oxidative stress and lipid peroxidation-derived DNA-lesions in inflammation driven carcinogenesis. Cancer Detect Prev, 2004, 28(6): 385-391 PMID: 15582261

[31] ChungFL, ChenHJ, NathRG. Lipid peroxidation as a potential endogenous source for the formation of exocyclic DNA adducts. Carcinogenesis, 1996, 17(10): 2105-2111 PMID: 8895475

[32] ZarkovicK. 4-hydroxynonenal and neurodegenerative diseases. Mol Aspects Med, 2003, 24(4–5): 293-303 PMID: 12893007

[33] WestJD, MarnettLJ. Endogenous reactive intermediates as modulators of cell signaling and cell death. Chem Res Toxicol, 2006, 19(2): 173-194 PMID: 16485894

[34] FengZ, HuW, TangMS. Trans-4-hydroxy-2-nonenal inhibits nucleotide excision repair in human cells: a possible mechanism for lipid peroxidation-induced carcinogenesis. Proc Natl Acad Sci USA, 2004, 101(23): 8598-8602 PMCID: 423240 PMID: 15187227

[35] BartschH, NairJ. Chronic inflammation and oxidative stress in the genesis and perpetuation of cancer: role of lipid peroxidation, DNA damage, and repair. Langenbecks Arch Surg, 2006, 391(5): 499-510 PMID: 16909291