Association Between HDL Functions and Atherosclerotic Burden in Healthy Individuals

This is a study of HDL function in healthy individuals classified in three groups according to their HDL-cholesterol levels (Low HDL-C, Intermediate HDl-C and High HDL-C), with the purpose of investigating which characteristics of the HDL particle might be associated to atherosclerotic burden, characterized by carotid intima-media thickness above 1mm.

Study Overview

• Status: Completed
• Conditions: Low HDL Cholesterol

Detailed Description

Selection: Study participants are initially selected trough the evaluation of consecutive lipid profiles from individuals who spontaneously seek governmental primary care centers of the city of Campinas, Sao Paulo, Brazil. Telephone-based screening interviews are performed, followed by in-person clinical evaluation and blood exams.

Biochemical analyses: Glucose, triglycerides, HDL-C and c-reactive protein (CRP) are measured in an automated Modular® Analytics Evo (Roche Diagnostics, Burgess Hill, West Sussex, UK), using Roche Diagnostics® reagents (Mannheim, Germany). LDL-cholesterol is calculated by the Friedewald formula. Serum insulin levels are measured using ELISA (Millipore, Massachusetts, USA). The Homeostasis Model Assessment (HOMA) Calculator version 2.2 (University of Oxford, UK) is used to estimate insulin sensitivity. Apolipoproteins A-I and B-100 and lipoprotein (a) are determined by nephelometry in an automated system and reagents from Dade-Behring® (Marburg, Germany).

Carotid artery ultrasound: Carotid artery atherosclerosis is estimated by using high-resolution B-mode ultrasound, at the posterior wall of the common carotid artery.

Lipoprotein isolation: HDL from each study participant is isolated from plasma, through ultracentrifugation (Beckman Coulter Inc., Palo Alto, USA).

HDL chemical composition: Using commercially available enzymatic kits, HDL content of total proteins, total cholesterol, free cholesterol, phospholipids, triglycerides and apolipoprotein A-I are measured. Cholesteryl ester (CE) is calculated as the difference between total cholesterol and free cholesterol times 1.67. HDL molar concentration is estimated based on particle total mass and molecular weight.

HDL physical-chemical characterization: HDL particle size is determined using dynamic light scattering, and zeta potential using laser Doppler micro-electrophoresis.

Determination of proteins involved in HDL metabolism: Cholesteryl ester transfer protein, phospholipids transfer protein, lipoprotein lipase, hepatic lipase and lecithin:cholesterol acyl transferase activities are determined trough radiometric exogenous assays.

HDL functions: Cholesterol efflux capacity, antioxidant activity, susceptibility to oxidation, anti-inflammatory activity and platelet aggregation inhibition are measured using straightforward and consolidated methodologies.

Statistical Analyses: Differences between groups are evaluated using ANOVA or Kruskal-Wallis, with Bonferroni's post-hoc multiple comparison analysis, according to variable distribution. Chi-Square is used to compare categorical data. Analysis of covariance (ANCOVA) adjusted for confounding variables is also used, after checking variables with histograms, normality plots, and residual scatter plots that tested for linearity, normality, and variance. Pearson's correlation test is used to assess the relationships between variables. A two-sided p-value of 0.05 is considered significant.

• Study Type: Observational
• Enrollment (Actual): 101

Contacts and Locations

Study Locations

• Brazil
  • Sao Paulo
    • Campinas, Sao Paulo, Brazil, 13083-887
      • State University of Campinas

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 20 years to 75 years (Adult, Older Adult)
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: All

• Sampling Method: Non-Probability Sample

Study Population

Individuals who sought governmental primary care centers of the city of Campinas, SP, Brazil.

Description

Inclusion Criteria:

• glucose <100 mg/dL
• Urea <71 mg/dL
• Creatinine < 1.20 mg/dL
• Uric acid <7.0 mg/dL
• Alanine aminotransferase <50 U/L
• Aspartate aminotransferase < 33 U/L
• Gamma-glutamyl transferase <71 U/L
• Alkaline phosphatase <129 U/L
• Thyroid stimulating hormone between 0.41 and 4.50 μUI/mL
• Free thyroxin between 0.9 and 1.8 ng/dL

Exclusion Criteria:

• Symptomatic cardiovascular disease
• LDL-cholesterol ≥130 mg/dL
• Triglycerides ≥150 mg/dL
• Metabolic syndrome
• BMI ≥ 30 kg/m2
• Smoking habit
• Daily intake of alcohol >14g
• Regular use of medical treatments

Study Plan

How is the study designed?

Number of groups / cohorts

3

Cohorts and Interventions

Group / Cohort
Low HDL-C; Study participants with HDL-C levels below the 10th percentile (HDL-C ≤32 mg/dL)
Intermediate HDL-C; Study participants with HDL-C levels between 40th and 60th percentiles (40≤HDL-C≤67 mg/dL)
High HDL-C; Study participants with HDL-C levels above the 90th percentile (HDL-C ≥78mg/dL)

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Cholesterol efflux capacity of HDL	This is a cross-sectional study and as such the cholesterol efflux capacity will be measured using the plasma sample collected at admission into the study	The analysis will be performed using plasma obtained at admission into the study

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Antioxidant activity of HDL	This is a cross-sectional study and as such the cholesterol efflux capacity will be measured using the plasma sample collected at admission into the study	The analysis will be performed using plasma obtained at admission into the study
Susceptibility to oxidation of HDL	This is a cross-sectional study and as such the cholesterol efflux capacity will be measured using the plasma sample collected at admission into the study	The analysis will be performed using plasma obtained at admission into the study
Anti-inflammatory activity of HDL	This is a cross-sectional study and as such the cholesterol efflux capacity will be measured using the plasma sample collected at admission into the study	The analysis will be performed using plasma obtained at admission into the study
Platelet aggregation inhibition by HDL	This is a cross-sectional study and as such the cholesterol efflux capacity will be measured using the plasma sample collected at admission into the study	The analysis will be performed using plasma obtained at admission into the study

Collaborators and Investigators

• Sponsor: University of Campinas, Brazil
• Collaborators: Fundação de Amparo à Pesquisa do Estado de São Paulo; Conselho Nacional de Desenvolvimento Científico e Tecnológico
• Investigators: Principal Investigator: Andrei C Sposito, PhD MD, University of Campinas, Brazil

Study record dates

Study Major Dates

• Study Start: April 1, 2008
• Primary Completion (Actual): November 1, 2013
• Study Completion (Actual): March 1, 2014

Study Registration Dates

• First Submitted: March 30, 2014
• First Submitted That Met QC Criteria: April 2, 2014
• First Posted (Estimate): April 7, 2014

Study Record Updates

• Last Update Posted (Estimate): April 7, 2014
• Last Update Submitted That Met QC Criteria: April 2, 2014
• Last Verified: April 1, 2014

More Information

Terms related to this study

• Keywords: HDL; Carotid artery intima-media thickness; Cholesterol efflux capacity; Antioxidant effect of HDL; Susceptibility to oxidation of HDL; Anti-inflammatory effect of HDL; Platelet aggregation inhibition by HDL
• Other Study ID Numbers: BHS2