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Methylation of the PPARg Promoter Region in Pregnancy

17. marts 2015 opdateret af: Hugo Mendieta Zeron, Materno-Perinatal Hospital of the State of Mexico

Methylation Evaluation of the PPARg Promoter Region (-351 to -260) in Pregnancy

The main objective of this study was to assess whether clinical, anthropometric, and biochemical variables of the mother were associated with changes in the methylation of the PPARg promoter region (-351 to -260). Methodology: This was a matched cohort study with two groups: a) normal weight (NW) pregnant women (n = 21) and their offspring, and b) overweight (OW) pregnant women (n = 20) and their offspring. DNA was extracted from leukocytes (4000-10,000 cells) in the MagnaPure (Roche) using the MagNAPure LC DNA Isolation Kit 1 (Roche, Germany). The treatment of DNA (2 µg) was performed with sodium bisulfite (EZ DNA Methylation-Direct Kit, ZymoResearch). Real-time polymerase chain reaction (qPCR) was performed in a LightCycler 2.0 (Roche) using the SYBR® Advantage® qPCR Premix Kit (Clontech).

Studieoversigt

Status

Afsluttet

Betingelser

Detaljeret beskrivelse

Women were recruited in the first trimester of pregnancy not including cases with congenital heart and disabling or autoimmune diseases. Those whose clinical follow-up were lost or, if in the postpartum period, who had to be attended in the obstetric intensive care unit were eliminated from the study.

A clinical visit per month was established. Body weight and height were measured in an overnight fasting status using an adult scale (Seca, Hamburg, Germany). Prepregnancy Body Mass Index (BMI) was calculated as weight in kg divided by height in meters squared based on the prenatal chart or on the self-reported weight of women with no prenatal chart.

Blood pressure was recorded at each visit using a standard sphygmomanometer (Riester Big Ben® Square, Germany). Preeclampsia was diagnosed and classified according to the American College of Obstetricians and Gynecologists (ACOG).

Fasting blood samples (10 ml) were taken at the HMPMP laboratory in an early morning after an overnight fasting. Serum samples were analyzed for glucose and lipid profile (Dimension Rx L Max, Dade Behring, USA). At the end of pregnancy, 1 to 2 ml of neonatal peripheral blood sample for leukocyte DNA extraction was taken.

Dietetic treatment was calculated according to height, weeks of gestation, and weight, considering an energy intake of 30 kcal/kg of ideal weight and a macronutrient distribution of: 55-65% carbohydrates, 10-20% fat, and the remainder as proteins. On each nutritional visit, the Healthy Eating Index for Pregnancy (HEI) was evaluated, and all women were recommended to include methionine-rich foods (beans, eggs, fish, garlic, lentils, onion, and soy) and those containing folic acid and vitamin B12 (beef liver, cereals, whole grains, yeast, etc.) in adequate quantities in their diet. The information was complemented with the Food Frequency Questionnaire (FFQ), and the diet adherence was considered adequate with 80% compliance to the indicated calories, at least in four visits.

This project had no risk to pregnant women and their infants, according to the regulations of the General Health Research Law of Mexico. We followed the Declaration of Helsinki, and all patients were asked to sign the written informed consent.

Undersøgelsestype

Observationel

Tilmelding (Faktiske)

41

Deltagelseskriterier

Forskere leder efter personer, der passer til en bestemt beskrivelse, kaldet berettigelseskriterier. Nogle eksempler på disse kriterier er en persons generelle helbredstilstand eller tidligere behandlinger.

Berettigelseskriterier

Aldre berettiget til at studere

18 år til 42 år (Voksen)

Tager imod sunde frivillige

Ja

Køn, der er berettiget til at studere

Kvinde

Prøveudtagningsmetode

Ikke-sandsynlighedsprøve

Studiebefolkning

Pregnant women attended at the Maternal-Perinatal Hospital "Mónica Pretelini Sáenz".

Beskrivelse

Inclusion Criteria: Women attended in the first trimester of pregnancy -

Exclusion Criteria: congenital heart and disabling or autoimmune diseases

-

Studieplan

Dette afsnit indeholder detaljer om studieplanen, herunder hvordan undersøgelsen er designet, og hvad undersøgelsen måler.

Hvordan er undersøgelsen tilrettelagt?

Design detaljer

Kohorter og interventioner

Gruppe / kohorte
Normal weight (NW) pregnant women

Pregnant women with Body Mass Index (BMI) > = 30. Body weight and height were measured in an overnight fasting status using an adult scale (Seca, Hamburg, Germany). Prepregnancy BMI was calculated as weight in kg divided by height in meters squared based on the prenatal chart or on the self-reported weight of women with no prenatal chart.

Dietetic treatment was calculated according to height, weeks of gestation, and weight, considering an energy intake of 30 kcal/kg of ideal weight and a macronutrient distribution of: 55-65% carbohydrates, 10-20% fat, and the remainder as proteins.

Overweight (OW) pregnant women

Pregnant women with Body Mass Index (BMI) < 30. Body weight and height were measured in an overnight fasting status using an adult scale (Seca, Hamburg, Germany). Prepregnancy BMI was calculated as weight in kg divided by height in meters squared based on the prenatal chart or on the self-reported weight of women with no prenatal chart.

Dietetic treatment was calculated according to height, weeks of gestation, and weight, considering an energy intake of 30 kcal/kg of ideal weight and a macronutrient distribution of: 55-65% carbohydrates, 10-20% fat, and the remainder as proteins.

Hvad måler undersøgelsen?

Primære resultatmål

Resultatmål
Foranstaltningsbeskrivelse
Tidsramme
Evidence of the effect of Body Mass Index (BMI) on the methylation status of the PPAR gamma promoter region (-351 to -260).
Tidsramme: Pregnant women were followed until delivery.

The treatment of DNA (2 µg) was performed with sodium bisulfite (EZ DNA Methylation-Direct Kit, ZymoResearch). For the control group, we used purified human methylated and unmethylated DNA (Zymo Research) with specific oligonucleotides. Lymphocyte DNA from healthy donors was used as negative control, and methylated DNA "in vitro" with Sss I enzyme (New England Biolabs) was used as positive control for methylation.

The methylated (M3) and unmethylated (U3) primers used were those proposed by Pancione et al.

Pregnant women were followed until delivery.

Samarbejdspartnere og efterforskere

Det er her, du vil finde personer og organisationer, der er involveret i denne undersøgelse.

Efterforskere

  • Ledende efterforsker: Hugo Mendieta Zerón, PhD., Asociación Científica Latina A.C.

Datoer for undersøgelser

Disse datoer sporer fremskridtene for indsendelser af undersøgelsesrekord og resumeresultater til ClinicalTrials.gov. Studieregistreringer og rapporterede resultater gennemgås af National Library of Medicine (NLM) for at sikre, at de opfylder specifikke kvalitetskontrolstandarder, før de offentliggøres på den offentlige hjemmeside.

Studer store datoer

Studiestart

1. september 2009

Primær færdiggørelse (Faktiske)

1. august 2010

Studieafslutning (Faktiske)

1. marts 2011

Datoer for studieregistrering

Først indsendt

11. marts 2015

Først indsendt, der opfyldte QC-kriterier

17. marts 2015

Først opslået (Skøn)

24. marts 2015

Opdateringer af undersøgelsesjournaler

Sidste opdatering sendt (Skøn)

24. marts 2015

Sidste opdatering indsendt, der opfyldte kvalitetskontrolkriterier

17. marts 2015

Sidst verificeret

1. marts 2015

Mere information

Begreber relateret til denne undersøgelse

Yderligere relevante MeSH-vilkår

Andre undersøgelses-id-numre

  • 05-06-2009

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