race: Caucasian gender: male age: 43 years disease status: healthy, obese tissue: white adipose treatment: resveratrol subject: 2
Treatment protocol
The design of the study was a randomized cross-over design with two different treatments: placebo and resVida (150mg/day trans-resveratrol [99.9%; provided by DSM Nutritional Products, Ltd.]) with a 4-week washout period. Subjects were instructed to take the first supplement on the day after baseline measurements (d1) and the last supplement in the evening on day 29. In the morning of day 30, after 36h in the respiration chamber, a microdialysis test was conducted to measure adipose tissue and skeletal muscle lipolysis before and for 6 hours after ingestion of a high-fat liquid test meal. At the end of the microdialysis test, a subcutaneous adipose tissue biopsy was taken.
Growth protocol
The current work was conducted as part of a previously published randomized double-blind crossover study (Timmers et al., Cell Metab 2011 Nov 2;14(5):612-22. PMID: 22055504 (GSE32357)). Nine healthy obese men (age 45–65 years; BMI 30-35 kg/m2) participated in a double-blind randomized cross-over study. Exclusion criteria were unstable body weight (weight gain or loss > 3 kg in the last three months), total body fat (%) < 25%, fasting plasma glucose > 6.1 mmol/l, hemoglobin < 7.8 mmol/l, engagement in programmed exercise > 2 hours total per week, impaired kidney and/ or hepatic function, first- or second-degree family member with type 2 diabetes mellitus, any medical condition requiring treatment and/or medication use, intake of dietary supplements except vitamins and minerals, unwilling to restrict high-resveratrol-containing foods, current alcohol consumption > 20 grams/day, participation in another biomedical study within 1 month before the first screening visit, a contraindication to MRI scanning. All subjects were informed about the nature of the study and written informed consent was obtained before study participation. The local Medical Ethical Committee of Maastricht University Medical Centre approved the study protocol.
Extracted molecule
total RNA
Extraction protocol
Adipose tissue biopsies were taken in the morning of day 30 after an overnight fast. Biopsies were obtained, immediately frozen in isopentane at its melting point, and stored at -80C until analysis. Total RNA was extracted from frozen adipose tissue specimens using TRIzol reagent (Invitrogen, Breda, the Netherlands) and purified on columns using the Qiagen RNeasy Micro Kit (Qiagen, Venlo, Netherlands). RNA quantity was measured with the ND-100 spectrophotometer (Isogen Life Science, Ijsselstein, the Netherlands), and RNA integrity was analysed on an Agilent 2100 BioAnalyzer (Agilent Technologies, Amsterdam, The Netherlands) using nanochips according to manufacturer's instructions.
Label
biotin
Label protocol
One hundred nanogram of RNA was used for whole transcript cDNA synthesis with the Ambion WT expression kit [catalog number 4411974] (Applied Biosystems/Life Technologies, Nieuwekerk a/d IJssel, The Netherlands).
Hybridization protocol
Hybridization and washing of the Affymetrix GeneChip Human Gene 1.1 ST peg arrays were performed on a GeneTitan Instrument according to the manufacturer's recommendations.
Scan protocol
Arrays were scanned on an Affymetrix GeneTitan instrument (Affymetrix, Santa Clara, CA).
Description
A09_03_2_1_Res
Data processing
Expression estimates were calculated applying the RMA algorithm in the Bioconductor library 'Oligo' (v1.20.4).
