J Lipid Atheroscler.  2016 Dec;5(2):145-154. 10.12997/jla.2016.5.2.145.

Impaired Antioxidant Ability of HDL and More Oxidized LDL are associated with Male Patients with Atrial Fibrillation

• Kim SJ
• Shin DG
• Kim JR
• Cho KH

• KMID: 2366139
• DOI: http://doi.org/10.12997/jla.2016.5.2.145

Abstract

OBJECTIVE
Atrial fibrillation (AF) is a common arrhythmia which is generally more prevalent in the elderly population. However, the new onset of AF is frequently found in young populations. In order to identify putative prognostic biomarkers for detection of young-onset AF, we purified and characterized lipoproteins in terms of oxidative and inflammatory properties.
METHODS
Male patients with AF (46±7 years of age, n=19) were recruited. Their serum and individual lipoproteins were analyzed and compared with healthy controls (48±9 years of age, n=17).
RESULTS
The patients with AF revealed hypertriglyceridemia, hyperuricemia with mild obesity, elevated levels of CRP, and a normal level of cholesterol. All lipoproteins from patients with AF demonstrated higher levels of TG and advanced glycated end products, and decreased particle size than controls. AF-LDL showed an increased extent of oxidation with increased atherogenic macrophage phagocytosis. AF-HDL showed impaired antioxidant ability and a lower level of apoA-I expression.
CONCLUSION
These data suggest that lipoprotein properties are severely modified in young AF patients, which were correlated with increased oxidation and inflammation.

Keyword

Atrial fibrillation; Lipoprotein; Oxidation; Apolipoproteins; Triacylglycerol

MeSH Terms

Aged
Apolipoprotein A-I
Apolipoproteins
Arrhythmias, Cardiac
Atrial Fibrillation*
Biomarkers
Cholesterol
Humans
Hypertriglyceridemia
Hyperuricemia
Inflammation
Lipoproteins
Macrophages
Male*
Obesity
Particle Size
Phagocytosis
Triglycerides
Apolipoprotein A-I
Apolipoproteins
Biomarkers
Cholesterol
Lipoproteins
Triglycerides

Figure

• 

  Fig. 1 Antioxidant ability of serum. (A) Serum paraoxonase activity. Twenty microliters of equally diluted serum (10 mg/mL) was added to 230µL of the paraoxon-ethyl (Sigma catalog# D-9286) solution containing 90 mM Tris-HCl, 3.6 mM NaCl, and 2 mM CaCl2 (pH 8.5) for 30 min. Error bars indicate the SD from three independent experiments with duplicate samples, (B) Ferric ion removal ability. Rate of increase in absorbance at 593 nm. The HDL3 of control group showed much weaker reducing ability than the HDL3 of control groups during 20 minutes of incubation.

• 

  Fig. 2 Comparison of oxidized extent in lipoprotein between the AF and control. (A) Quantification of conjugated diene level in lipoproteins under the same protein amount (VLDL, 3µg; LDL, 6µg; HDL2, 6µg; HDL3, 10 mg), (B) Determination of thiobarbituric acid reactive substances between AF and control groups using malondialdehyde standard. The same amount of protein was used. VLDL, 0.15 mg/mL; LDL, 0.33 mg/mL, (C) Monitoring of conjugated diene level production in LDL (0.3 mg/mL) under presence of Cu2+ (final 10µM), (D) Electromobility of LDL from the AF and controls with or without treatment of copper (final 10µM) for 6 hrs (0.5% agarose gel)

• 

  Fig. 3 Quantification of advanced glycated end products. The extent of glycation was determined from fluorescence (Ex=370 nm, Em=440 nm) measurement in each lipoprotein fraction using the same amount of protein. VLDL, 15µg; LDL, 32µg; HDL2, 30µg; HDL3, 50µg.

• 

  Fig. 4 Expressional profiles of apolipoproteins in HDL. (A) Electrophoretic patterns of HDL2 from AF and control group (15% SDS-PAGE). Purified HDL2 via ultracentrifugation was equally diluted and the same amount of protein (3µg) was loaded per lane and visualized by Coomassie blue staining, (B) mmunodetection of apoA-I in HDL2 and HDL3. Western blotting analysis with apoA-I antibody, which was raised by full-length apoA-I (ab7613; Abcam).

• 

  Fig. 5 Cellular uptake of VLDL and LDL into macrophage cell. PMA differentiated THP-1 cells were incubated with 50µL of VLDL (0.15 mg/mL) or LDL (0.33 mg/mL) from either AF or control group under presence of 450µL of RPMI1640 media. The extent of cellular uptake of the lipoproteins by macrophages was then compared by oil red O staining as described in the text. The cells were then photographed using a Nikon Eclipse TE2000 microscope (Tokyo, Japan) at 400× magnification.

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