Background: Obesity in pregnancy can contribute to epigenetic changes. Aim: To assess whether body mass index (BMI) in pregnancy is associated with changes in the methylation of the peroxisome proliferatorâÂ€Â‘activated receptor γ (PPAR) promoter region (−359 to − 260) in maternal and neonatal leukocytes. Subjects and Methods: In this matched, cohort study 41 pregnant women were allocated into two groups: (a) Normal weight (n = 21) and (b) overweight (n = 20). DNA was extracted from maternal and neonatal leukocytes (4000–10,000 cells) in MagNA Pure (Roche) using MagNA Pure LC DNA Isolation Kit 1 (Roche, Germany). Treatment of DNA (2 μg) was performed with sodium bisulfite (EZ DNA MethylationâÂ€Â‘Direct™ Kit; Zymo Research). RealâÂ€Â‘time quantitative polymerase chain reaction (qPCR) was performed in a LightCycler 2.0 (Roche) using the SYBR® Advantage® qPCR Premix Kit (Clontech). The primers used for PPARγ coactivator (PPARG) M3 were 5’âÂ€Â‘aagacggtttggtcgatcâÂ€Â‘3’ (forward), and5’âÂ€Â‘cgaaaaaaaatccgaaatttaaâÂ€Â‘3’ (reverse) and those for PPARG unmethylated were: 5’âÂ€Â‘gggaagatggtttggttgattâÂ€Â‘3’ (forward) and 5’âÂ€Â‘ttccaaaaaaaaatccaaaatttaaâÂ€Â‘3’ (reverse). Intergroup differences were calculated using the Mann–Whitney UâÂ€Â‘test, and intragroup differences, with the Wilcoxon test (IBM SPSS Statistics for Windows, Version 19.0. Armonk, NY: IBM Corp.). Results: Significant differences were found in BMI, pregestational weight, and postdelivery weight between groups but not in the methylation status of the PPARγ promoter region (−359 to − 260). Conclusion: The PPARγ promoter region (−359 to − 260) in peripheral leukocytes is unlikely to get an obesityâÂ€Â‘induced methylation in pregnancy.
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