Pardo F, Silva L, Sáez T, Salsoso R, Gutiérrez J, Sanhueza C, Leiva A, Sobrevia L
Cellular and Molecular Physiology Laboratory (CMPL), Division of Obstetrics and Gynaecology, School of Medicine, Faculty of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile.
1] Cellular and Molecular Physiology Laboratory (CMPL), Division of Obstetrics and Gynaecology, School of Medicine, Faculty of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile [2] Cellular Signalling and Differentiation Laboratory (CSDL), Faculty of Health Sciences, Universidad San Sebastián, Santiago, Chile.
Int J Obes (Lond). 2015 Aug;39(8):1264-73. doi: 10.1038/ijo.2015.57. Epub 2015 Apr 14.
Human foetal development and growth in an environment of maternal obesity associates with high risk of cardiovascular disease and adverse neonatal outcome. We studied whether supraphysiological gestational weight gain results in human fetoplacental endothelial dysfunction and altered fetoplacental vascular reactivity.
Primary cultures of human umbilical vein endothelial cells (HUVECs) and umbilical vein rings were obtained from pregnant women (112 total of patients recruited, 7 patients dropped out) exhibiting prepregnancy normal weight that ended with a physiological (pGWG (n=67), total weight gain 11.5-16 kg, rates of weight gain ≤0.42 kg per week) or supraphysiological (spGWG (n=38), total weight gain >16 kg, rates of weight gain >0.42 kg per week) gestational weight gain (reference values from US Institute of Medicine guidelines). Vascular reactivity to insulin (0.1-1000 nmol l(-1), 5 min) in KCl-preconstricted vein rings was measured using a wire myograph. Protein levels of human equilibrative nucleoside transporter 1 (hENT1), total and Ser(1177)- or Thr(495)-phosphorylated endothelial nitric oxide synthase (eNOS) were detected by western blot or immunofluorescence, and adenosine transport (0-250 μmol l(-1) adenosine, 2 μCi ml(-1) [(3)H]adenosine, 20 s, 25 °C) was measured in the presence or absence of 1 μmol l(-1) nitrobenzylthioinosine (hENT1 inhibitor) or 10 μmol l(-1) chlorpromazine (CPZ, endocytosis inhibitor) in HUVECs.
spGWG associates with reduced NOS activity-dependent dilation of vein rings (P=0.001), lower eNOS expression and higher Thr(495) (P=0.044), but unaltered Ser(1177)eNOS phosphorylation. hENT1-adenosine maximal transport activity was reduced (P=0.041), but the expression was increased (P=0.001) in HUVECs from this group. CPZ increased hENT1-adenosine transport (P=0.040) and hENT1 plasma membrane accumulation only in cells from pGWG.
spGWG in women with a normal prepregnancy weight causes lower fetoplacental vascular reactivity owing to the downregulation of eNOS activity and adenosine transport in HUVECs. Maternal spGWG is a detrimental condition for human fetoplacental endothelial function and reducing these alterations could result in a better neonate outcome.
在母体肥胖环境下的人类胎儿发育和生长与心血管疾病高风险及不良新生儿结局相关。我们研究了超生理妊娠体重增加是否会导致人胎盘胎儿内皮功能障碍及胎盘胎儿血管反应性改变。
从孕前体重正常且妊娠体重增加为生理水平（pGWG，n = 67，总体重增加11.5 - 16 kg，体重增加速率≤0.42 kg/周）或超生理水平（spGWG，n = 38，总体重增加>16 kg，体重增加速率>0.42 kg/周）（参考美国医学研究所指南的数值）的孕妇中获取人脐静脉内皮细胞（HUVECs）原代培养物和脐静脉环（共招募112例患者，7例退出）。使用线肌张力测定仪测量KCl预收缩静脉环对胰岛素（0.1 - 1000 nmol·l⁻¹，5分钟）的血管反应性。通过蛋白质印迹法或免疫荧光法检测人平衡核苷转运体1（hENT1）、总内皮型一氧化氮合酶（eNOS）以及Ser(1177)-或Thr(495)-磷酸化eNOS的蛋白水平，并在HUVECs中存在或不存在1 μmol·l⁻¹硝基苄硫肌苷（hENT1抑制剂）或10 μmol·l⁻¹氯丙嗪（CPZ，内吞作用抑制剂）的情况下测量腺苷转运（0 - 250 μmol·l⁻¹腺苷，2 μCi·ml⁻¹ [³H]腺苷，20秒，25°C）。
spGWG与静脉环中一氧化氮合酶（NOS）活性依赖性扩张降低相关（P = 0.001），eNOS表达降低且Thr(495)升高（P = 0.044），但Ser(1177)eNOS磷酸化未改变。该组HUVECs中hENT1 - 腺苷最大转运活性降低（P = 0.041），但表达增加（P = 0.001）。CPZ仅在pGWG组细胞中增加hENT1 - 腺苷转运（P = 0.040）和hENT1质膜积累。
孕前体重正常的女性发生spGWG会导致胎盘胎儿血管反应性降低，这是由于HUVECs中eNOS活性和腺苷转运下调所致。母体spGWG对人胎盘胎儿内皮功能是一种有害状况，减少这些改变可能会带来更好的新生儿结局。
