Methylation Evaluation of the PPARg Promoter Region (-351 to -260) in Pregnancy
试验速览
- 阶段
- 不适用
- 状态
- 已完成
- 发起方
- 入组人数
- 41
- 主要终点
- Evidence of the effect of Body Mass Index (BMI) on the methylation status of the PPAR gamma promoter region (-351 to -260).
研究概览
简要总结
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).
详细描述
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.
研究设计
- 研究类型
- Observational
- 观察模型
- Cohort
- 时间视角
- Prospective
入排标准
- 年龄范围
- 18 Years 至 42 Years(Adult)
- 性别
- Female
- 接受健康志愿者
- 是
入选标准
- •Women attended in the first trimester of pregnancy -
排除标准
- •congenital heart and disabling or autoimmune diseases
结局指标
主要结局
Evidence of the effect of Body Mass Index (BMI) on the methylation status of the PPAR gamma promoter region (-351 to -260).
时间窗: 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.
次要结局
未报告次要终点
研究者
Hugo Mendieta Zeron
Médico Internista adscrito a la Unidad de Cuidados Intensivos Obstétricos
Materno-Perinatal Hospital of the State of Mexico
