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Cell-free Biochemical Fluorometric Enzymatic Assay for High-throughput Measurement of Lipid Peroxidation in High Density Lipoprotein
Published on: October 12, 2017
Circulating oxidized LDL, increased in patients with acute myocardial infarction, is accompanied by heavily modified
Naoko Sawada1, Takashi Obama1, Shinji Koba2
1Division of Biological Chemistry, Department of Pharmaceutical Sciences Showa University School of Pharmacy, Shinagawa-ku, Tokyo 142-8555, Japan.
Insights
This study reveals two types of in vivo-oxidized LDL (oxLDL) in human plasma. Electronegative oxLDL, found in acute myocardial infarction patients, is associated with oxidized HDL particles.
Area of Science:
- Cardiovascular Science
- Lipid Metabolism
- Atherosclerosis Research
Background:
- Oxidized LDL (oxLDL) is a key factor in the development of atherosclerosis.
- Understanding the structural characteristics of circulating oxLDL is crucial for assessing cardiovascular risk.
Purpose of the Study:
- To identify structural features of in vivo-oxidized LDL (oxLDL) in human circulation.
- To investigate the relationship between oxLDL and atherosclerosis.
Main Methods:
- Fractionation of LDL using anion-exchange chromatography.
- Detection of in vivo-oxLDL using anti-oxidized PC (oxPC) monoclonal antibody (mAb).
- Analysis of LDL fractions using transmission electron microscopy and Western blotting.
Main Results:
- In vivo-oxLDL was found in flow-through and electronegative LDL [LDL(-)] fractions.
- Electronegative in vivo-oxLDL levels were threefold higher in acute myocardial infarction patients compared to healthy subjects.
- The LDL(-) fraction contained apoA1 and apoB, with HDL-sized particles and acrolein adducts on apoA1.
Conclusions:
- Two distinct types of in vivo-oxLDL exist in human plasma.
- Electronegative in vivo-oxLDL is associated with oxidized HDL and may play a significant role in atherogenesis.
Abstract:
Oxidized LDL (oxLDL) is a known risk factor for atherogenesis. This study aimed to reveal structural features of oxLDL present in human circulation related to atherosclerosis. When LDL was fractionated on an anion-exchange column, in vivo-oxLDL, detected by the anti-oxidized PC (oxPC) mAb, was recovered in flow-through and electronegative LDL [LDL(-)] fractions. The amount of the electronegative in vivo-oxLDL, namely oxLDL in the LDL(-) fraction, present in patients with acute MI was 3-fold higher than that observed in healthy subjects. Surprisingly, the LDL(-) fraction contained apoA1 in addition to apoB, and HDL-sized particles were observed with transmission electron microscopy. In LDL(-) fractions, acrolein adducts were identified at all lysine residues in apoA1, with only a small number of acrolein-modified residues identified in apoB. The amount of oxPC adducts of apoB was higher in the LDL(-) than in the L1 fraction, as determined using Western blotting. The electronegative in vivo-oxLDL was immunologically purified from the LDL(-) fraction with an anti-oxPC mAb. The majority of PC species were not oxidized, whereas oxPC and lysoPC did not accumulate. Here, we propose that there are two types of in vivo-oxLDL in human circulating plasma and the electronegative in vivo-oxLDL accompanies oxidized HDL.
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