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Identification of Oxidized Phosphatidylinositols Present in OxLDL and Human Atherosclerotic Plaque

  • Original Article
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Lipids

Abstract

Oxidized low-density lipoprotein (OxLDL) plays an important role in initiation and progression of atherosclerosis. Proatherogenic effects of OxLDL have been attributed to bioactive phospholipids generated during LDL oxidation. It is unknown what effect oxidation has on the phosphatidylinositol (PtdIns) molecules in LDL, even though PtdIns is 6% of the total LDL phospholipid pool. We sought to identify and quantitate oxidized phosphatidylinositol (OxPtdIns) species in OxLDL and human atherosclerotic plaque. Bovine liver PtdIns was subjected to non-enzymatic and lipoxygenase-catalyzed oxidation. Reversed-phase liquid chromatography with negative ESI–MS identified and confirmed compounds by fragmentation pattern analysis from which an OxPtdIns library was generated. Twenty-three OxPtdIns molecules were identified in copper-oxidized human LDL at 0, 6, 12, 24, 30, and 48 h, and in human atherosclerotic plaque. In OxLDL, OxPtdIns species containing aldehydes and carboxylates comprised 17.3 ± 0.1 and 0.9 ± 0.2%, respectively, of total OxPtdIns in OxLDL at 48 h. Hydroperoxides and isoprostanes at 24 h (68.5 ± 0.2 and 22.8 ± 0.2%) were significantly greater than 12 h (P < 0.01) without additional changes thereafter. Hydroxides decreased with increased oxidation achieving a minimum at 24 h (5.2 ± 0.3%). Human atherosclerotic plaques contained OxPtdIns species including aldehydes, carboxylates, hydroxides, hydroperoxides and isoprostanes, comprising 18.6 ± 4.7, 1.5 ± 0.7, 16.5 ± 7.4, 33.3 ± 1.1 and 30.2 ± 3.3% of total OxPtdIns compounds. This is the first identification of OxPtdIns molecules in human OxLDL and atherosclerotic plaque. With these novel molecules identified we can now investigate their potential role in atherosclerosis.

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Abbreviations

CM:

2:1 Chloroform:methanol

BHT:

Butylated hydroxytoluene

EDTA:

Ethylenediaminetetraacetic acid

EPD:

Embolic protection device

IL:

Interleukin

I/R:

Ischemia/reperfusion

KDdiA-PtdCho:

1-Palmitoyl-2-(4-keto-dodec-3-ene-dioyl)-phosphatidylcholine

LGE2:

Levuglandin E2

LOX-I:

Lipoxygenase-I

Lp-PLA2:

Lipoprotein-associated phospholipase A2

MRM:

Multiple reaction monitoring

NP:

Normal-phase

OxLDL:

Oxidized low density lipoprotein

OxPtdCho:

Oxidized phosphocholine

OxPtdIns:

Oxidized phosphatidylinositol

PAF:

Platelet-activating factor

PAPtdCho:

1-Palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine

PCI:

Percutaneous coronary intervention

PDHPtdCho:

1-Palmitoyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine

PEI:

1-Palmitoyl-2-epoxyisoprostane

PGPtdCho:

1-Palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine

PtdIns:

Phosphatidylinositol

PtdIns-4-P:

Phosphatidylinositol 4-phosphate

PtdIns-4,5-P2 :

Phosphatidylinositol 4,5-bisphosphate

PtdIns-3,4,5-P3 :

Phosphatidylinositol 3,4,5-trisphosphate

PtdIns 3K:

Phosphoinositide 3-kinases

PKB:

Protein kinase B

PLPtdCho:

1-Palmitoyl-2-linoleoyl-sn-glycero-3-phosphocholine

PONPtdCho:

1-Palmitoyl-2-(9-oxo)nonanoyl-sn-glycero-3-phosphocholine

POVPtdCho:

1-Palmitoyl-2-(5-oxo-valeroyl)-sn-glycero-3-phosphocholine

ROS:

Reactive oxygen species

RP:

Reversed-phase

SAPtdCho:

1-Stearoyl-2-arachidonoyl-sn-glycero-3-phosphocholine

SEC:

1-Stearoyl-epoxycholine

SEI:

1-Stearoyl-epoxyisoprostane

SLPtdCho:

1-Stearoyl-2-linoleoyl-sn-glycero-3-phosphocholine

SVG:

Saphenous vein graft

TIC:

Total ion chromatogram

TNFα:

Tumor necrosis factor alpha

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Acknowledgments

This research was supported by operating grants from the Heart and Stroke foundation of Canada and Research Manitoba. DH was a recipient of a Research Manitoba graduate scholarship.

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Correspondence to Amir Ravandi.

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Hasanally, D., Edel, A., Chaudhary, R. et al. Identification of Oxidized Phosphatidylinositols Present in OxLDL and Human Atherosclerotic Plaque. Lipids 52, 11–26 (2017). https://doi.org/10.1007/s11745-016-4217-y

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