treatment: placebo subjectid: 229 before/after supplementation: before
Treatment protocol
The study was a randomized, double blind, placebo-controlled, cross-over trial in which subjects received two 4-week interventions; epicatechin (100mg/day) or placebo (microcrystalline cellulose). Details of the study have been previously described (Dower et al., 2015; PMID: 25934864). In brief, subjects received both interventions in random order with a wash-out period of 4-week between both interventions. Subjects consumed 2 capsules per day with a glass of water: one during breakfast and one during dinner. Subjects were asked to avoid consumption of epicatechin-rich foods throughout the study. Fasting peripheral blood mononuclear cells (PBMC) were collected before (T0) and after each intervention period (T4) using BD Vacutainer Cell Preparation Tubes. On the day prior to each study day, subjects received a standardized evening meal, refrained from alcohol consumption and strenuous exercise, and were not allowed to eat or drink anything except water after 10.00 pm.
Growth protocol
Apparently healthy men and women aged 40–80 y were recruited from Wageningen and surroundings, in The Netherlands. Eligible for participation were nonsmoking men and women, with a BMI (in kg/m2) between 20 and 40 and systolic blood pressure (SBP) between 125 and 160 mm Hg on 2 separate occasions. Exclusion criteria included the presence of diabetes mellitus; cardiovascular, gastrointestinal, or liver disease; usage of antihypertensive or cholesterol-lowering medication or corticosteroids; adherence to a prescribed diet; unstable weight in the past 2 mo; ≥10 h/wk moderate to vigorous physical activity (≥3 metabolic equivalent tasks); and pregnancy or lactation. All subjects gave written informed consent for participation in this study, which was reviewed and approved by the local Medical Ethical Committee of Wageningen University. All procedures were executed according to the declaration of Helsinki. This study was registered at clinicaltrials.gov as NCT01691404.
Extracted molecule
total RNA
Extraction protocol
RNA was isolated (RNeasy Micro kit, Qiagen, Venlo, the Netherlands), quantified (Nanodrop ND 1000, Nanodrop technologies, Wilmington, Delaware USA) and integrity was checked by an Agilent 2100 Bioanalyser with RNA 6000 microchips (Agilent Technologies, South Queensferry, UK). Samples were included for microarray analysis if the RNA integrity number (RIN) was > 7.
Label
biotin
Label protocol
One hundred nanogram of total RNA was used for whole transcript cDNA synthesis with the Whole-Transcript Sense Target Assay (Affymetrix, Santa Clara, CA, USA; P/N 900652).
Hybridization protocol
Samples were hybridized to Affymetrix GeneChip Human Gene 1.1 ST array plates (Santa Clara, CA) according to manufacturer's instructions.
Scan protocol
Arrays were scanned on an Affymetrix GeneTitan instrument (Affymetrix, Santa Clara, CA).
Data processing
Expression estimates were calculated applying the RMA algorithm in the Bioconductor library 'Oligo' (v1.36.1).
Pure epicatechin and inflammatory gene expression profiles in circulating immune cells in (pre) hypertensive adults; a randomized double-blind, placebo-controlled, crossover trial
