High-density Lipoprotein Subclasses Research Articles

Effect of Colesevelam HCl Monotherapy on Lipid Particles in Type 2 Diabetes Mellitus

PurposeIn addition to lowering hemoglobin A1C, colesevelam has been shown to improve the atherogenic lipoprotein profile of subjects with type 2 diabetes mellitus (T2DM) when used in combination with metformin and/or sulfonylureas. A recent study evaluated the effects of colesevelam as antidiabetes monotherapy in adults with T2DM who had inadequate glycemic control (hemoglobin A1C ≥7.5 to ≤9.5 %) with diet and exercise alone; we report here the effects on lipoprotein particle subclasses.MethodsSubjects were randomized to receive oral colesevelam 3.75 g/day (n = 176) or placebo (n = 181) for 24 weeks. Changes in lipoprotein particle subclasses were determined by nuclear magnetic resonance spectroscopy.ResultsAt Week 24 with last observation carried forward, colesevelam produced a reduction in total low-density lipoprotein (LDL) particle concentration (baseline: 1,611 nmol/L; least-squares [LS] mean treatment difference: −143 nmol/L, p < 0.0001) versus placebo; reductions were also seen in large, small, and very small LDL particle concentrations (all p < 0.05). There was also a reduction in total very low-density lipoprotein (VLDL) and chylomicron particle concentration (baseline: 88 nmol/L; LS mean treatment difference: −1 nmol/L, p = 0.82) that resulted from a lowering in small VLDL particle concentration (baseline: 45 nmol/L; LS mean treatment difference: −5 nmol/L, p = 0.03). In addition, with colesevelam there was an increase in total high-density lipoprotein (HDL) particle concentration versus placebo (baseline: 31 μmol/L; LS mean treatment difference: +0.6 μmol/L, p = 0.20), due to increases in the large (baseline: 5 μmol/L; LS mean treatment difference: +0.5 μmol/L, p = 0.007) and medium (baseline: 3 μmol/L; LS mean treatment difference: +0.8 μmol/L, p = 0.02) HDL subclasses.ConclusionsColesevelam monotherapy in subjects with T2DM resulted in generally favorable changes in certain lipoprotein subclass profiles compared with placebo.

A Closer Look at Cardioprotective Function of HDL: Revise the HDL – Cholesterol Hypothesis?

BACKGROUND: The strong inverse association of plasma levels of high-density lipoprotein (HDL) cholesterol with coronary heart disease (CHD) found in human epidemiological studies led to the development of the ‘HDL cholesterol hypothesis’, which posits that intervention to raise HDL cholesterol will result in reduced risk of CHD. A number of recent developments have brought the potential protective role of HDL into question. Several clinical trials of agents that substantially raise HDL-C have been demonstrated to not reduce CHD event rates.CONTENT: For decades, HDL and HDL-cholesterol (HDL-C) levels were viewed as synonymous, and modulation of HDL-C levels by drug therapy held great promise for the prevention and treatment of cardiovascular disease. Nevertheless, recent failures of drugs that raise HDL-C to reduce cardiovascular risk and the now greater understanding of the complexity of HDL composition and biology have prompted researchers in the field to redefine HDL. As such, the focus of HDL has now started to shift away from a cholesterol-centric view toward HDL particle number, subclasses, and other alternative metrics of HDL. Many of the recently discovered functions of HDL are, in fact, not strictly conferred by its ability to promote cholesterol flux but by the other molecules it transports, including a diverse set of proteins, small RNAs, hormones, carotenoids, vitamins, and bioactive lipids. Based on HDL’s ability to interact with almost all cells and deliver fat-soluble cargo, HDL has the remarkable capacity to affect a wide variety of endocrine-like systems.SUMMARY: There is a significant need to redefine HDL and its benefit. HDL transports a diverse set of functional proteins, including many binding proteins. HDL transports and deliver vitamins, carotenoids, and other small molecules. Moreover, HDL transports hormones, steroids and bile acids, and can modulate multiple endocrine pathways. HDLs also transport and deliver microRNAs to recipient cells and control gene expression. Likewise, HDLs carry bioactive lipids and can activate signaling cascades and receptors that control endothelial apoptosis, migration, survival and activation. Many of HDL’s alternative noncholesterol cargo likely confer many of HDL’s alternative functions.KEYWORDS: HDL, ApoA1, RCT, ABCA1, ABCG1, miRNA, HDL lipidome, HDL proteome

Effects of Lipid-Lowering Drugs on High-Density Lipoprotein Subclasses in Healthy Men—A Randomized Trial

Context and ObjectiveInvestigating the effects of lipid-lowering drugs on HDL subclasses has shown ambiguous results. This study assessed the effects of ezetimibe, simvastatin, and their combination on HDL subclass distribution.Design and ParticipantsA single-center randomized parallel 3-group open-label study was performed in 72 healthy men free of cardiovascular disease with a baseline LDL-cholesterol of 111±30 mg/dl (2.9±0.8 mmol/l) and a baseline HDL-cholesterol of 64±15 mg/dl (1.7±0.4 mmol/l). They were treated with ezetimibe (10 mg/day, n = 24), simvastatin (40 mg/day, n = 24) or their combination (n = 24) for 14 days. Blood was drawn before and after the treatment period. HDL subclasses were determined using polyacrylamide gel-tube electrophoresis. Multivariate regression models were used to determine the influence of treatment and covariates on changes in HDL subclass composition.ResultsBaseline HDL subclasses consisted of 33±10% large, 48±6% intermediate and 19±8% small HDL. After adjusting for baseline HDL subclass distribution, body mass index, LDL-C and the ratio triglycerides/HDL-C, there was a significant increase in large HDL by about 3.9 percentage points (P<0.05) and a decrease in intermediate HDL by about 3.5 percentage points (P<0.01) in both simvastatin-containing treatment arms in comparison to ezetimibe. The parameters obtained after additional adjustment for the decrease in LDL-C indicated that about one third to one half of these effects could be explained by the extent of LDL-C-lowering.ConclusionsIn healthy men, treatment with simvastatin leads to favorable effects on HDL subclass composition, which was not be observed with ezetimibe. Part of these differential effects may be due to the stronger LDL-C-lowering effects of simvastatin.Trial RegistrationClinicalTrials.gov NCT00317993

Less but better: cardioprotective lipid profile of patients with GCK-MODY despite lower HDL cholesterol level

Patients with diabetes caused by single-gene mutations generally exhibit an altered course of diabetes. Those with mutations of the glucokinase gene (GCK-MODY) show good metabolic control and low risk of cardiovascular complications despite paradoxically lowered high-density lipoprotein (HDL) cholesterol levels. In order to investigate the matter, we analyzed the composition of low-density lipoprotein (LDL) and HDL subpopulations in such individuals. The LipoPrint© system (Quantimetrix, USA) based on non-denaturing, linear polyacrylamide gel electrophoresis was used to separate and measure LDL and HDL subclasses in fresh-frozen serum samples from patients with mutations of glucokinase or HNF1A, type 1 diabetes (T1DM) and healthy controls. Fresh serum samples from a total of 37 monogenic diabetes patients (21 from GCK-MODY and 16 from HNF1A-MODY), 22 T1DM patients and 15 healthy individuals were measured in this study. Concentrations of the small, highly atherogenic LDL subpopulation were similar among the compared groups. Large HDL percentage was significantly higher in GCK-MODY than in control (p = 0.0003), T1DM (p = 0.0006) and HNF1A-MODY groups (p = 0.0246). Patients with GCK-MODY were characterized by significantly lower intermediate HDL levels than controls (p = 0.0003) and T1DM (p = 0.0005). Small, potentially atherogenic HDL content differed significantly with the GCK-MODY group showing concentrations of that subfraction from control (p = 0.0096), T1DM (p = 0.0193) and HNF1A-MODY (p = 0.0057) groups. Within-group heterogeneity suggested the existence of potential gene–gene or gene–environment interactions. GCK-MODY is characterized by a strongly protective profile of HDL cholesterol subpopulations. A degree of heterogeneity within the groups suggests the existence of interactions with other genetic or clinical factors.

Evidence for the presence of active paraoxonase 1 in small-dense low-density lipoprotein

AimWe have recently demonstrated the quick ex vivo transfer of paraoxonase 1 (PON1) activity from high-density lipoprotein (HDL) to small, dense low-density lipoprotein (sdLDL). We set out to assess whether sdLDL contains active PON1 in vivo.MethodsWe conducted a nested case–control, proof of principle study with the Japanese healthy subjects with normal lipids (n = 23) and age and gender-paired dyslipidemic subjects (n = 17). Lipid panels, lactonase and arylesterase assays, and PON1 zymogram in the LDL and HDL subclasses were assessed.ResultsPON1 specific activity in the high-molecular weight lipoprotein fraction corresponding to LDL migration was found in 48% of normo and in 29% of dyslipidemic Japanese subjects. This band co-localizes with apoB100 and not Lp(a) and displays a lower molecular mass than the bulk of LDL.ConclusionWe provide evidence, for the first time, that native sdLDL contains up to 4% of the total PON1 activity in the serum of up to 48% of the Japanese subjects. Could the PON1-containing sdLDL represent a set of particles with a defense mechanism from oxidation and therefore its levels actually prove to be atheroprotective? If further studies confirm this contention, a zymogram of PON1 in LDL subclasses could be a functional assay that complements the current methods that only inform on the size and lipid concentration of these particles.

Comparison of four methods of analysis of lipoprotein particle subfractions for their association with angiographic progression of coronary artery disease

BackgroundCompare gradient gel electrophoresis (GGE), vertical auto profile ultracentrifugation (VAP-II), nuclear magnetic resonance spectroscopy (NMR), and ion mobility for their ability to relate low- (LDL), intermediate- (IDL), very-low-density (VLDL) and high-density lipoprotein (HDL) subfraction concentrations to atherosclerotic progression. Methods and resultsRegression analyses of 136 patients who received baseline and follow-up coronary angiographies and subfraction measurements by all four methods in the HDL Atherosclerosis Treatment Study. Prior analyses have shown that the intervention primarily affected disease progression in proximal arteries with <30% stenoses at baseline.Three-year increases in percent stenoses were consistently associated with higher on-study plasma concentrations of small, dense LDL as measured by GGE (LDLIIIb, P = 10−6; LDLIVa, P = 0.006; LDLIVb, P = 0.02), VAP-II (LDL4, P = 0.002), NMR (small LDL, P = 0.001), and ion mobility (LDL IIb, P = 0.04; LDLIIIa, P = 0.002; LDLIIIb, P = 0.0007; LDLIVa, P = 0.05). Adjustment for triglycerides, HDL-cholesterol, and LDL-cholesterol failed to eliminate the statistical significance for on-study GGE estimated LDLIIIb (P = 10−5) and LDLIVa (P = 0.04); NMR-estimated small LDL (P = 0.03); or ion mobility estimated large VLDL (P = 0.02), LDLIIIa (P = 0.04) or LDLIIIb (P = 0.02). All methods show that the effects were significantly greater for the on-study than the baseline small, dense LDL concentrations, thus establishing that the values concurrent to the progression of disease were responsible. The rate of disease progression was also related to individual VLDL, IDL, and HDL subclasses to differing extents among the various analytic methods. ConclusionFour methodologies confirm the association of small, dense LDL with greater coronary atherosclerosis progression, and GGE, NMR, and ion mobility confirm that the associations were independent of standard lipid measurements.Clinical Trial Registration: clinicaltrials.gov/ct2/show/NCT00000553.

The Value and Distribution of High-Density Lipoprotein Subclass in Patients with Acute Coronary Syndrome

BackgroundHigh-density lipoprotein (HDL) enhances cholesterol efflux from the arterial wall and exhibits potent anti-inflammatory and anti-atherosclerosis (AS) properties. Whether raised HDL levels will clinically benefit patients with acute coronary syndrome (ACS) and the value at which these effects will be apparent, however, is debatable. This study examined the HDL subclass distribution profile in patients with ACS.MethodsPlasma HDL subclasses were measured in 158 patients with established ACS and quantified by two-dimensional gel electrophoresis and immunoblotting. ACS diagnosis was based on symptoms of cardiac ischemia, electrocardiogram (ECG) abnormalities, speciality cardiac enzyme change along with presence of coronary heart disease (CHD) on coronary angiography.ResultsThe small-sized preβ1-HDL, HDL3b, and HDL3a levels were significantly higher, and the large-sized HDL2a and HDL2b levels were significantly lower in patients with ACS than in those with stable angina pectoris (SAP) and in normal control subjects. Meanwhile, with an elevation in the low-density lipoprotein cholesterol (LDL-C), fasting plasma glucose (FPG), body mass index (BMI), and blood pressure (BP), and the reduction in the high density lipoprotein cholesterol (HDL-C) levels, the HDL2b contents significantly decreased and the preβ1-HDL contents significantly increased in patients with ACS. The correlation analysis revealed that the apolipoprotein (apo)A-I levels were positively and significantly with all HDL subclasses contents; plasma total cholesterol (TC) and fasting plasma glucose (FPG) levels were inversely associated with HDL2a, and HDL2b. Moreover, the FPG levels were positively related to HDL3c, HDL3b, and HDL3a in ACS patients.ConclusionThe HDL subclass distribution profile remodeling was noted in the patients with ACS. Plasma lipoprotein and FPG levels, BP, and BMI play an important role in the HDL subclass metabolism disorder for patients with ACS. The HDL subclass distribution phenotype might be useful as a novel biomarker to assist in the risk stratification of patients with ACS.

Paraoxonase 1 lactonase activity and distribution in the HDL subclasses in the cord blood

ObjectivesParaoxonase 1 (PON1) is a lactonase with important antioxidant and immunoprotective properties. We hypothesized that PON1 lactonase activity, PON1, and high-density lipoprotein (HDL) subclasses distribution are different in neonates than in adults.Material and methodsWe studied 83 healthy term neonates (34 males and 49 females) who were born by spontaneous, uncomplicated vaginal delivery. The study also included 17 paired maternal blood samples as well as 20 non-pregnant women collected for comparison. Total and free PON1 lactonase and arylesterase activity, HDL subclasses, PON1, and apolipoprotein distribution in the subclasses were assayed.ResultsPON1 arylesterase activity in the cord blood represented 37% ± 4 of the maternal activity, whereas the PON1 lactonase activity amounted to only 23% ± 5 of the maternal activity. The free arylesterase and lactonase activities were higher in the cord blood by 16 and 36%, respectively. There is a 65% lower HDL2b PON1 in the cord blood than in the maternal serum. When the Lipoprint HDL subclasses were assayed, the neonates showed a larger content (52% higher) of very large HDL as well as a characteristic peak in the middle-sized HDL5 which is unremarkable in the mothers.ConclusionThe novel findings of this study are that the neonates have lower PON1 lactonase activity, higher free PON1, different distributions of PON1 in the HDL subclasses as compared with their mother and adults as well as a distinctive HDL subclass lipid profile. Our data also suggest that the neonate HDL is enriched with an intermediate-sized (and/or less charged HDL) that is also rich in active PON1.

HIV/hepatitis C virus coinfection ameliorates the atherogenic lipoprotein abnormalities of HIV infection

Higher levels of small low-density lipoprotein (LDL) and lower levels of high-density lipoprotein (HDL) subclasses have been associated with increased risk of cardiovascular disease. The extent to which HIV infection and HIV/hepatitis C virus (HCV) coinfection are associated with abnormalities of lipoprotein subclasses is unknown. Lipoprotein subclasses were measured by nuclear magnetic resonance (NMR) spectroscopy in plasma samples from 569 HIV-infected and 5948 control participants in the Fat Redistribution and Metabolic Change in HIV Infection (FRAM), Coronary Artery Risk Development in Young Adults (CARDIA), and Multi-Ethnic Study of Atherosclerosis (MESA) studies. Multivariable regression was used to estimate the association of HIV and HIV/HCV coinfection with lipoprotein measures with adjustment for demographics, lifestyle factors, and waist-to-hip ratio. Relative to controls, small LDL levels were higher in HIV-monoinfected persons (+381 nmol/l, P <0.0001), with no increase seen in HIV/HCV coinfection (-16.6 nmol/l). Levels of large LDL levels were lower (-196 nmol/l, P <0.0001) and small HDL were higher (+8.2 μmol/l, P < 0.0001) in HIV monoinfection with intermediate values seen in HIV/HCV coinfection. Large HDL levels were higher in HIV/HCV-coinfected persons relative to controls (+1.70 μmol/l, P <0.0001), whereas little difference was seen in HIV-monoinfected persons (+0.33, P = 0.075). Within HIV-infected participants, HCV was associated independently with lower levels of small LDL (-329 nmol/l, P <0.0001) and small HDL (-4.6 μmol/l, P <0.0001), even after adjusting for demographic and traditional cardiovascular risk factors. HIV-monoinfected participants had worse levels of atherogenic LDL lipoprotein subclasses compared with controls. HIV/HCV coinfection attenuates these changes, perhaps by altering hepatic factors affecting lipoprotein production and/or metabolism. The effect of HIV/HCV coinfection on atherosclerosis and the clinical consequences of low small subclasses remain to be determined.

Interrelated cathepsin S-lowering and LDL subclass profile improvements induced by atorvastatin in the plasma of stable angina patients.

We hypothesized that, in stable angina patients, atorvastatin therapy lowers the cathepsin S (CTSS) concentrations, as assessed non-invasively according to a plasma analysis. In addition, the low-density lipoprotein (LDL) and high-density lipoprotein (HDL) size and subclasses in the plasma were analysed to establish the association between CTSS and lipoprotein metabolism and determine whether this association is atorvastatin-sensitive. A total of 43 patients with stable angina received atorvastatin therapy (20 mg/day, 10 weeks). The plasma CTSS mRNA levels, CTSS protein concentrations and CTSS activity, as well as LDL and HDL size and subclasses, were analysed before and after treatment. Atorvastatin treatment did not change the plasma CTSS mRNA levels, although it lowered the plasma CTSS concentrations and activity. An increased plasma CTSS concentration and activity were found to be associated with a more atherogenic LDL subclass profile (a decreased dominant LDL size and increased percentage of small, dense LDL particles). The atorvastatin-induced CTSS-loweringeffect was concomitant with an improvement in the LDL subclass profile, and the changes were found to be interrelated. Concomitant, interrelated changes in the CTSS levels and LDL subclass profiles were found in the LDL phenotype B patients only (a dominant LDL diameter of ≤ 25.5 nm at the start of the study). In this subgroup, lowering of the plasma CTSS mRNA level also correlated with lowering of the proportion of small, dense LDL particles. Atorvastatin-induced CTSS-lowering and LDL subclass profile improvements in the plasma of LDL phenotype B patients with stable angina are concomitant and interrelated.

Distribution of Low-Density Lipoprotein and High-Density Lipoprotein Subclasses in Patients With Sarcoidosis

Systemic inflammatory diseases are associated with proatherogenic lipoprotein profile, but there is a lack of information regarding overall distributions of lipoprotein subclasses in sarcoidosis. To investigate whether patients with sarcoidosis have altered distributions of plasma low-density lipoprotein (LDL) and high-density lipoprotein (HDL) particles. Seventy-seven patients with biopsy-proven sarcoidosis (29 with acute and 48 with chronic sarcoidosis) treated with corticosteroids and 77 age- and sex-matched controls were included in the study. Low-density lipoprotein and HDL subclasses were determined by gradient gel electrophoresis, while inflammatory markers and lipid parameters were measured by standard laboratory methods. Compared to controls, patients had fewer LDL I subclasses (P < .001), but more LDL II and III (P < .001) subclasses. This pattern was evident in both acute and chronic disease groups. Patients also had smaller HDL size (P < .001) and higher proportions of HDL 2a (P = .006) and 3a particles (P = .004). Patients with chronic sarcoidosis had smaller LDL size than those with acute disease (P = .02) and higher proportions of HDL 3a subclasses (P = .04) than controls. In acute sarcoidosis, relative proportions of LDL and HDL particles were associated with levels of inflammatory markers, whereas in chronic disease an association with concentrations of serum lipid parameters was found. The obtained results demonstrate adverse lipoprotein subfraction profile in sarcoidosis with sustained alterations during disease course. Evaluation of LDL and HDL particles may be helpful in identifying patients with higher cardiovascular risk, at least for prolonged corticosteroid therapy due to chronic disease course.

Association of glutathione-S-transferase gene polymorphism and lipoprotein subclasses in hemodialysis patients

ObjectivesEnd-stage renal disease (ESRD) is characterized by profound dyslipidemia and enhanced oxidative stress. The patients also show evidence of exhausted and/or deficient anti-oxidative defense enzymes, one of them being glutathione-S-transferase (GST). This study investigates relationship between GST gene polymorphism and low-density lipoprotein (LDL) and high-density lipoprotein (HDL) subclasses in ESRD. Design and methodsGSTM1, T1, and P1 genotypes were determined by polymerase chain reaction–restriction fragment length polymorphism in 160 patients undergoing hemodialysis. LDL and HDL subclasses were separated by gradient gel electrophoresis and biochemical parameters were measured by routine laboratory methods. ResultsGSTM1-positive patients had higher proportion of small, dense LDL III particles than those with GSTM1-null genotype (P<0.05). Similarly, GSTP1-Ile/Ile patients had higher proportion of LDL III (P<0.05), but more HDL 2b and less HDL 3a particles than GSTP1-Ile/Val and Val/Val carriers (P<0.05). LDL subclass distribution in smokers with GSTM1-null genotype was shifted towards smaller particles, as compared to GSTM1-positive and GSTM1-null non-smokers. Smokers with GSTP1-Ile/Val and Val/Val genotypes had smaller LDL size than their non-smoking counterparts (P<0.05). Both smokers and non-smokers with GSTP1 Ile/Ile genotype had more LDL III particles than non-smokers carrying Val allele. Non-smokers with GSTP1 Ile/Ile genotype had more HDL 2b subclasses than non-smokers with GSTP1-Ile/Val and Val/Val (P<0.05), but less HDL 3a particles than smokers with GSTP1-Ile/Val and Val/Val genotypes (P<0.05). GSTT1 gene polymorphism had no effect on lipoprotein subclass distributions. ConclusionsOur results demonstrate significant associations between low activity GST genotypes and proatherogenic lipoprotein particles in hemodialysis patients which might further increase their cardiovascular disease risk.

Activation of paraoxonase 1 is associated with HDL remodeling ex vivo

BackgroundWe hypothesize that during high density lipoprotein (HDL) remodeling PON1 reaches an optimal distribution in HDL subclasses by which it achieves maximum activity. We conducted this study to gain insight on PON1 fate and activation during short-term HDL remodeling ex vivo. MethodsSerum from 8 healthy volunteers was either frozen at −80°C (time 0) or incubated under sterile conditions for up to 48h at 37°C or at 4°C. Aliquots were taken at 3, 6, 9, 24 and 48h and immediately frozen at −80°C. PON1 activities were measured, as well as PON1 and apolipoprotein distributions in HDL subclasses by gradient gel electrophoresis. ResultsThe first novel finding in our study is the evidence provided for a significant activation of both lactonase and arylesterase activities of PON1 that ensues in a very short time frame of incubation of serum ex vivo at 37°C. All subjects studied displayed these changes, the activation was apparent in <3h, peaked at 6h and amounted to >20%. This is associated with a temperature and time-dependent redistribution of PON1 activity in HDL subclasses, with an increase in activity in both very large HDL2 and small HDL3 in the first phase (3–9h), followed by a progressive transfer of PON1 to very large HDL2 as the particles mature. These changes are paralleled by the appearance of weak, but apparent PON1 activity at subspecies that correspond to sdLDL. During the first phase of PON1 activation and shifts, a parallel shift of apoE can be evidenced: at 3–9h, apoE increases in sdLDL, after that time it is lost from HDL and also from sdLDL and stays in VLDL at the origin of the run. ApoA-I shifts towards larger particles, which parallels the change in PON1. As HDL matures there is a progressive shift of apoA-II towards larger HDL. Low levels of apoA-IV at the initiation of the incubation are followed by time dependent quick disappearance of apoA-IV in HDL which parallels the changes in PON1, apoE and A-II. ConclusionShort, ex vivo incubation of serum leads to quick activation of PON1 associated with transfers to HDL3c, large HDL and sdLDL. The process is blocked by CETP and LCAT inhibitors. The data suggest that HDL maturation optimizes PON1 activity. These findings may be of interest for future studies aimed at modulating PON-1 activity for its cardioprotective effects and suggest a new mechanism whereby CETP inhibitors failed in clinical trials.

High-Density Lipoprotein Particle Subclass Heterogeneity and Incident Coronary Heart Disease

Raising the cholesterol of high-density lipoprotein (HDL) particles is targeted as a cardiovascular disease prevention strategy. However, HDL particles are heterogeneous in composition and structure, which may relate to differences in antiatherogenic potential. We prospectively evaluated the association of HDL subclasses, defined by a recently proposed nomenclature, with incident coronary heart disease (CHD). Baseline HDL particle concentrations were measured by nuclear magnetic resonance spectroscopy and categorized into 5 subclasses (very large, large, medium, small, and very small) among 26 332 initially healthy women. During a median follow-up of 17 years, 969 cases of incident CHD (myocardial infarction, revascularization, and CHD death) were ascertained. In Cox models that adjusted for age, race/ethnicity, blood pressure, smoking, postmenopausal status, and hormone therapy, associations with incident CHD were inverse (P trend<0.0001) for concentrations of very large (hazard ratio for top versus bottom quartile, 0.49; 95% confidence interval, 0.41-0.60), large (0.54; 0.45-0.64), and medium (0.69; 0.58-0.83) HDL subclasses. Conversely, hazard ratios (95% confidence intervals) for small and very small HDL were 1.22 (1.01-1.46; P trend=0.08) and 1.67 (1.39-2.02; P trend<0.0001), respectively. However, after additionally adjusting for metabolic and lipoprotein variables, associations for the spectrum of large, medium, and small HDL subclasses were inverse (P trend<0.05 for large and small and 0.07 for medium), whereas subclasses at either end of the spectrum were not associated with CHD (P trend=0.97 for very large and 0.21 for very small HDL). In this prospective study, associations with incident CHD differed by HDL particle subclass, which may be relevant for developing HDL-modulating therapies. URL: http://www.clinicaltrials.gov. Unique identifier: NCT00000479.

Serum cholesterol and triglyceride reference ranges of twenty lipoprotein subclasses for healthy Japanese men and women

AimThis epidemiological study was done to generate normal ranges for the cholesterol and triglyceride levels in serum lipoprotein subclasses isolated from healthy adults based on gender and menopausal status. MethodsCholesterol and triglyceride levels in 20 lipoprotein subclasses as separated by high performance liquid chromatography were measured in serum obtained from 825 fasting healthy subjects (267 men, 558 women). ResultsFor serum cholesterol, 13.7% was found in very low density lipoprotein (VLDL) subclasses, 55.6% in low density lipoprotein (LDL) subclasses, and 30.4% in high density lipoprotein (HDL) subclasses. For serum triglycerides, these values were 52.1%, 27.9%, and 17.4%, respectively. Levels of cholesterol in some VLDL subclasses were inversely correlated with the levels of some HDL subclasses, while for triglycerides, elevated levels in any one subclass were generally strongly associated with elevated levels in all other subclasses. Men had significantly higher large VLDL-cholesterol levels than women (P < 0.05), while women had significantly higher small VLDL-cholesterol levels than men (P < 0.001). Women had significantly higher large LDL- and large and medium HDL-cholesterol levels than men (P < 0.001). Men had significantly higher chylomicron (CM), large and medium VLDL-, and small LDL-triglyceride levels than women (P < 0.001). Women had significantly higher very large and large HDL-triglyceride levels than men (P < 0.01). Postmenopausal women had significantly higher CM, all VLDL, and large, medium and small LDL-cholesterol levels, and significantly higher all VLDL, LDL, and HDL-triglyceride levels than premenopausal women (P < 0.001). ConclusionsOur data document important gender and menopausal status differences in cholesterol and triglyceride subclass levels, as well as significant correlations between values in the various serum lipoprotein subclasses.

Smaller HDL particles are associated with absence of obstructive coronary artery disease in stable angina pectoris patients

A research on novel cardiovascular risk factors is mainly focused on patients with clinically verified coronary artery disease (CAD), while less is known about their presence in symptomatic patients, but without angiographically proven occlusion of coronary arteries. The aim of this study was to compare plasma low-density lipoprotein (LDL) and high-density lipoprotein (HDL) size and subclasses in stable angina patients with and without significant obstructive CAD. LDL and HDL subclasses were analysed in 100 stable angina patients with ≥50% of obstruction and 40 patients with less than 50% of luminal narrowing, as assessed by coronary angiography. Patients with <50% of obstruction had reduced mean HDL size and higher proportion of small HDL particles (P < 0.05). HDL size and proportion of small HDL particles were significant and independent predictors of obstructive CAD (P < 0.05, respectively). Stable angina patients with <50% of coronary obstruction have more favourable HDL subclasses distribution than patients with significant coronary stenosis.

Impaired high-density lipoprotein anti-oxidant capacity in human abdominal aortic aneurysm

Abdominal aortic aneurysm (AAA) is a particular form of atherothrombotic disease characterized by the dilation of the aortic wall and the presence of an intraluminal thrombus (ILT). The objective of the present study was to evaluate the pro-oxidant properties of the ILT and to characterize the anti-oxidant capacity of high-density lipoproteins (HDLs). Our results show that ILT, adventitia, and plasma from AAA patients contained high concentrations of lipid and protein oxidation products. Mediators produced within or released by the thrombus and the adventitia were shown to induce reactive oxygen species (ROS) production by cultured aortic smooth muscle cells (AoSMCs) and to trigger the onset of apoptosis (an increase in mitochondrial membrane potential). Iron chelation limited these effects. Both concentration and functionality of HDLs were altered in AAA patients. Plasma levels of Apo A-I were lower, and small HDL subclasses were decreased in AAA patients. Circulating HDLs in AAA patients displayed an impaired capacity to inhibit copper-induced low-density lipoprotein oxidation and AoSMC ROS production. Western blot analyses of HDLs demonstrated that myeloperoxidase is associated with HDL particles in AAA patients. ILT and adventitia are a source of pro-oxidant products, in particular haemoglobin, which may impact on the wall stability/rupture in AAA. In addition, HDLs from AAA patients exhibit an impaired anti-oxidant activity. In this context, restoring HDL functionality may represent a new therapeutic option in AAA.

Abdominal adiposity is associated with high-density lipoprotein subclasses in Japanese schoolchildren

BackgroundThe large HDL subclass is considered to possess cardioprotective properties. The purpose of this study is to determine the relationship among abdominal adiposity, insulin resistance and HDL subclass profiles of Japanese schoolchildren. MethodsThe study subjects included 164 children (79 boys and 85 girls). We obtained waist to height ratio (WHtR), lipid profile, and HOMA-IR. The HDL subclass profile was analyzed by HPLC. ResultsChildren of either sex with abdominal obesity (WHtR≧0.5) had reduced concentrations of very large, large, and medium HDLC in conjunction with elevated triglyceride (TG) concentrations and HOMA-IR. WHtR was inversely related to the concentrations of very large (boys: r=−0.5306, p<0.0001; girls: r=−0.3483, p=0.0011), large (r=−0.6168, p<0.0001; r=−0.4387, p<0.0001), and medium (r=−0.4170, p=0.0001; r=−0.4116, p<0.0001) HDLC. The multiple regression analyses revealed that WHtR was an independent factor of the concentrations of very large, large, small, and very small HDLC in boys and the concentrations of large and medium HDLC in girls. ConclusionsIn Japanese schoolchildren, abdominal obesity is associated with atherogenic HDL subclass profile. Abdominal obesity may be an important target for the prevention and management of HDL subclass alteration, even in children who do not suffer from insulin resistance or hypertriglyceridemia.

A Single Amino Acid Substitution in the Group 1 Trypanosoma brucei gambiense Haptoglobin-Hemoglobin Receptor Abolishes TLF-1 Binding

Critical to human innate immunity against African trypanosomes is a minor subclass of human high-density lipoproteins, termed Trypanosome Lytic Factor-1 (TLF-1). This primate-specific molecule binds to a haptoglobin-hemoglobin receptor (HpHbR) on the surface of susceptible trypanosomes, initiating a lytic pathway. Group 1 Trypanosoma brucei gambiense causes human African Trypanosomiasis (HAT), escaping TLF-1 killing due to reduced uptake. Previously, we found that group 1 T. b. gambiense HpHbR (TbgHpHbR) mRNA levels were greatly reduced and the gene contained substitutions within the open reading frame. Here we show that a single, highly conserved amino acid in the TbgHpHbR ablates high affinity TLF-1 binding and subsequent endocytosis, thus evading TLF-1 killing. In addition, we show that over-expression of TbgHpHbR failed to rescue TLF-1 susceptibility. These findings suggest that the single substitution present in the TbgHpHbR directly contributes to the reduced uptake and resistance to TLF-1 seen in these important human pathogens.

Relationship between the distribution of plasma HDL subclasses and the polymorphisms of APOA5 in hypertriglyceridemia

AimsThis study aims to examine the possible associations between high density lipoprotein (HDL) subclass distribution and APOA5-1131T>C polymorphism in hypertriglyceridemia. MethodsThe distribution of HDL subclasses was quantified by 2-dimensional electrophoresis in conjunction with immunodetection method. The APOA5-1131T>C polymorphism was identified in 95 hypertriglyceridemic (HTG) patients and 102 healthy subjects by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). ResultsThe APOA5-1131C (C) allele frequency was higher in the HTG group than in the control group. Plasma triglycerides (TG) were significantly higher and apoA5 was significantly lower in patients with the C allele when compared to patients with the APOA5-1131T (T) allele, even more dramatically so in the APOA5-1131CC homozygote. In both the HTG group and the control group, the frequency of the C allele was positively correlated with levels of TG, total cholesterol (TC), low density lipoprotein cholesterol (LDL-C) and apolipoprotein B100 (apoB100), and negatively correlated with levels of high density lipoprotein cholesterol (HDL-C), apolipoprotein A1 (apoA1) and apolipoprotein A5 (apoA5) (P<0.001). In all subjects, the frequency of the C allele was positively correlated with the level of small-sized HDL (preβ1-HDL and HDL3a), and negatively correlated with levels of HDL2a and HDL2b. ConclusionChanges in HDL subclass distributions in HTG may be related to the APOA5-1131T>C polymorphism. This polymorphism leads to a general shift towards smaller-sized HDL.