Introduction Adequate maternal supply and placental delivery of long chain polyunsaturated fatty acids (LCPUFA) is essential for normal fetal development. supplementation increased placental uptake of DHA (P<0.05) while HFD alone had no measurable effect. Resveratrol increased AMP-activated protein kinase activity and mRNA expression of the fatty acid transporters FATP-4 CD36 and FABPpm (P<0.05). Placental DHA content was decreased in HFD dams; resveratrol had no effect on tissue fatty acid profiles. Discussion Maternal HFD did not significantly affect placental LCPUFA uptake. Furthermore resveratrol stimulated placental Rabbit polyclonal to F10. DHA uptake capacity AMPK activation and transporter expression. Placental handling of DHA is particularly sensitive to the dramatic alterations in the maternal metabolic phenotype and placental AMPK activity associated with resveratrol supplementation. fed a diet composed of 35% fat comparable to a typical western-style diet before and during pregnancy have offspring born with fatty liver who develop obesity and vascular dysfunction in later life [15 16 Characteristics of these pregnancies include maternal insulin resistance decreased uterine blood flow decreased plasma LCPUFA levels high placental triglycerides and placental inflammation [17] mimicking complications observed in pregnancies of obese women [18-20] and possibly altering placental nutrient handling. Resveratrol is a polyphenol that mimics calorie restriction and has been shown to improve outcomes in animal models of obesity [21-23]; effects in obese humans are more controversial [24-26]. Recently Roberts reported in the model that supplementation of a high fat diet (HFD) with resveratrol during pregnancy improves maternal fasting insulin levels and TAK-733 decreases fat mass in addition to resolving fetal outcomes such as high liver triglycerides [27]. Furthermore resveratrol infusions were shown to acutely increase uterine blood flow in this model of maternal high fat diet [27]. Resveratrol’s caloric restriction effects may depend upon activation of the AMP-activated protein kinase (AMPK) pathway [28] which as an energy sensor in turn stimulates numerous metabolic processes within the cell including fatty acid uptake and metabolism with the goal of increasing cellular energy production [29 30 AMPK activity is decreased in placentas of obese women [19]. The consequences of AMPK TAK-733 activation on placental fatty acid handling are unknown. We sought to determine the effect of HFD with and without resveratrol supplementation on placental fatty acid handling using the model. We hypothesized that a diet high in fat would suppress placental LCPUFA uptake and resveratrol supplementation would oppose this effect in association with the activation of the cellular energy sensor AMPK. Materials and Methods Experimental Design All animal procedures were in accordance with the guidelines of the Institutional Animal Care and Use Committee of the Oregon National Primate Research Center (ONPRC) and Oregon Health & Science University. were fed control chow (15% fat n=5) HFD (35% fat n=10) or HFD containing 0.37% resveratrol (n=5) prior to- and throughout pregnancy as previously described [27]. At ~130d gestation (term=173d) placentas were collected by caesarean section. placentas have two lobes: a primary lobe TAK-733 attached to the umbilical cord and a secondary lobe attached to the primary via interlobar vessels. Whether these lobes have different functional capacity is not well understood and therefore we collected samples (four separate cotyledons) from both lobes. Sets of samples were: 1) flash frozen immediately in liquid nitrogen and stored at ?80°C for molecular analyses; 2) formalin fixed for subsequent paraffin embedding for immunohistochemistry analyses; 3) collected fresh for uptake studies as described below. Fatty acid uptake Placental fatty acid uptake studies using 14C-labeled oleic acid (OA) arachidonic acid TAK-733 (AA) and docosahexanoic acid (DHA) were performed in placental explants as previously described [7]. Briefly villous tissue was collected from both placental lobes and washed in warm PBS. Small fragments (<2mm3) were dissected and one from each cotyledon was placed in each Netwell insert in warm uptake buffer (HBSS with 10mM HEPES pH 7.4) for 30 minute equilibration at 37°C. Buffer was removed and 200 μM albumin-bound fatty acids (ratio of 1 1:1 fatty acid:BSA) in uptake buffer.