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Glycosylation enhances oxygen radical-induced modifications and decreases acetylhydrolase activity of human low density lipoprotein

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Abstract

Background. Posttranslational nonenzymatic glycosylation of native low-density lipoprotein (n-LDL) occurs bothin vitro andin vivo in diabetic patients. Glycosylated LDL (glc-LDL) behave similarly to oxidized LDL in some respects. In fact, unlike n-LDL, uptake of glc-LDL can occur in part by the “scavenger” receptor(s), as also demonstrated for oxidized LDL. The enzyme acetylhydrolase, carried by LDL, catabolizes platelet activating factor (PAF). This enzymatic activity is inhibited in oxidized LDL. However, it is unknown whether glc-LDL have reduced acetylhydrolase activity.Objectives. The first aim of the study was to investigate whether glc-LDL were more susceptible than n-LDL to oxidative modification, and which different oxygen radical species were involved in the phenomenon. Moreover, in order to investigate whether glycosylation may affect acetylhydrolase, we also measured this enzymatic activity in both n- and glc-LDL.Methods. In vitro glc-LDL and n-LDL were exposed to the oxidants xanthine/xanthine oxidase (X/XO; 2 mM and 100 mU/ml, respectively), or CuSO4 (10 μM) for 18 hs at 37°C. Parallel experiments were done in the presence of the superoxide radical scavenger superoxide dismutase (SOD; 330 U/ml), the hydrogen peroxide scavenger catalase (1000 U/ml), or the hydroxyl radical scavenger dimethylthiourea (10 mM) or dimethylsulfoxide (1 mM). Standards of PAF and lyso-PAF were visualized with iodine vapors after separation by thin layer chromatography. The distribution of label was determined by an imaging scanner. Labeled products were then isolated from the chromatography plate, and the amount of3H-lyso-PAF formed was determined by liquid scintillation counting.Results. Glc-LDL were more susceptible than n-LDL to lipid peroxidation (n-LDL 22.9±3.4 vs 34.8±4.2* nmoles/MDA/mg of protein in glc-LDL oxidized by X/XO and n-LDL 28.9±4.2 vs 40.4±4.1* in glc-LDL oxidized by CuSO4,*p<0.05 vs n-LDL). SOD, but not other scavengers, prevented peroxidation, indicating an obligatory role for superoxide radicals. Oxidation of glc-LDL also induced a higher degree of apolipoprotein-B100 modifications than n-LDL, with increased electrophoresis mobility and decreased TNBS reactivity. These effects were similarly prevented by SOD. Finally, acetylhydrolase activity was significantly lower in glc-LDL than in n-LDL.Conclusion. Glycosylation increases LDL oxidation due to superoxide radicals, and also reduces acetylhydrolase activity. These phenomenona may contribute to enhance and/or accelerate the progression of atherosclerosis in diabetic patients.

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Abbreviations

LDL:

low density lipoprotein

n-LDL:

native LDL

glc-LDL:

glycosylated LDL

PAF:

platelet activating factor

X/XO:

xanthine/xanthine oxidase reaction

SOD:

superoxide dismutase

DMTU:

dimethylthiourea

DMSO:

dimethylsulfoxide

TNBS:

trinitrobenzenesulfonic acid

MDA:

malonyldialdehyde

LPO:

lipid peroxides

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Authors and Affiliations

  1. Via B. Falcomatà, 5, 80128, Naples, Italy

    C. Napoli

  2. Division of Clinical Immunology, Federico II University of Naples, Via S. Pansini 5, 80131, Naples, Italy

    M. Triggiani

  3. Department of Cellular and Molecular Pathology “L. Califano”, Federico II University of Naples, Via S. Pansini 5, 80131, Naples, Italy

    G. Palumbo

  4. Division of Cardiology Department of Medicine, Federico II University of Naples, Via S. Pansini, 5, 80131, Naples, Italy

    C. Napoli, M. Condorelli, M. Chiariello & G. Ambrosio

  5. Division of Cardiology, University of Perugia, Via Eugubina, 42, 06100, Perugia, Italy

    G. Ambrosio

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Napoli, C., Triggiani, M., Palumbo, G. et al. Glycosylation enhances oxygen radical-induced modifications and decreases acetylhydrolase activity of human low density lipoprotein. Basic Res Cardiol 92, 96–105 (1997). https://doi.org/10.1007/BF00805570

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  • DOI: https://doi.org/10.1007/BF00805570

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