Metabolic Syndrome Clinical Trial
Official title:
FTO rs9939609 and PPARy rs1801282 Polymorphisms in Mexican Adolescents With Overweight and Obesity at High Risk for Developing Diabetes
Background and Aims: The presence of the FTO rs9939609 and PPARy rs1801282 polymorphisms suggests changes in energy metabolism; this variation may be responsible for the development of various diseases including obesity. The aim of this study was to identify the presence of these polymorphisms in Mexican adolescents with overweight and obesity at high risk for developing diabetes. Methods and Results: This was a descriptive cross-sectional study, where 71 healthy adolescents (average age of 16) were included. Anthropometric measurements, Body mass index, as well as the determination of glucose, insulin and HOMA index were calculated from all the patients. The FTO rs9939609 and PPARy rs1801282 polymorphisms were determined by real-time PCR.
Study Population Students without known disease, between 14 and 18 years from the "Lic.
Adolfo Lopez Mateos" Preparatory school of the Autonomous University of the State of Mexico
(UAEMex), were invited to participate.
Anthropometry The body mass index (BMI) was calculated as weight (kg) divided by height (m)
squared with conventional stadiometer; dividing the results in normal, overweight and
obesity according to the classification of the World Health Organization (WHO), specifically
for adolescents by gender and age.
Laboratory 3 mL of venous blood (Vacutainer tubes) were taken in fasting. The concentration
of glucose, total cholesterol, HDL-cholesterol, LDL-cholesterol and triglycerides were
determined by enzymatic methods (Randox Laboratories Ltd, County Antrim, UK); whereas leptin
and adiponectin concentrations were determined by the immunosorbent assay (ELISA)
(Invitrogen MR Frederyck MD USA). A sample remained frozen at -70°C (900 series Thermo
Scientific) until DNA extraction.
Genotyping DNA extraction was performed in the Magna Pure LC 2.0 Instrument (Roche, Germany)
using the MagNA Pure LC DNA Isolation Kit 1 (Roche, Germany). After extraction, the DNA was
quantified using a NanoPhotometer (Implen GmbH, Germany), reporting concentration (μg/ml)
and purity (260/280 absorbance). Genotyping was performed by polymerase chain reaction (PCR)
Life express thermal cycler (Bioer, China). The primers and conditions for each polymorphism
are listed in Table 1.
The oligonucleotides were designed using the Primer Quest web tool (Integrated DNA
Technologies, Inc., CA, USA) and synthesized at the Synthesis and DNA Sequencing Unit of the
National Autonomous University of Mexico (UNAM) Institute of Biotechnology (Cuernavaca,
Morelos, Mexico). A search was performed in BLAST to verify the correct hybridization. The
final products were visualized in 2% agarose gels, stained with ethidium bromide, and
visualized under an UV transilluminator (Gel Logic 212 Pro, Carestream, USA). To confirm the
detection of polymorphisms sequencing was performed at the National Institute of Genomic
Medicine (INMEGEN), Mexico City.
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Observational Model: Ecologic or Community, Time Perspective: Prospective
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