Yang Xiaotao, Xu Ping, Zhang Fumei, Zhang Li, Zheng Yangxi, Hu Mingyu, Wang Lulu, Han Ting-Li, Peng Chuan, Wang Lianlian, Wen Li, Zeng Yiwen, Gao Rufei, Xia Yong, Tong Chao, Yang Zhu, Qi Hongbo, Baker Philip N
Department of Obstetrics, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
International Joint Laboratory of Reproduction and Development, Ministry of Education of China, Chongqing Medical University, Chongqing, China.
Cell Physiol Biochem. 2018;49(2):578-594. doi: 10.1159/000492995. Epub 2018 Aug 30.
BACKGROUND/AIMS: Preeclampsia (PE) has long been assumed to be an ischemic disease of the placenta, although there is limited evidence as to how the ischemia impacts on the placenta. AMP-activated protein kinase (AMPK) is a key regulator of cellular energy metabolism and plays an important role in a variety of ischemic diseases by enhancing energy production. The present study investigated placental metabolism in PE, and the role of AMPK in regulating trophoblast function.
placentas from normal and PE complicated pregnancies were subjected to GC-MS to identify fatty acids (FA) metabolic fingerprints, and total FA oxidation was assessed by malondialdehyde (MDA) measurement. The AMPK-ACC signaling pathway was assessed by q-PCR and Western Blotting. HTR8/SVneo trophoblast cultures were exposed to different oxygenation conditions to establish an in vitro PE cell model; further analysis by GC-MS for metabolite profiling was then undertaken. Trophoblasts invasion was assessed by a matrigel transwell assay in the presence/absence of AMPK expression and after manipulations of AMPK activity, and then further validated by human villi outgrowth experiments.
AMPK phosphorylation and MDA production were significantly elevated in placentas from pregnancies complicated by PE. Metabolism of cis double bond FA was inhibited while trans double bond FA metabolism was promoted in PE placentas. HTR8/SVneo cell culture conditions of persistent low oxygenation mimicked the hyper-activation of AMPK and enhanced the FA oxidation that was observed in PE. AMPK activation impaired trophoblast invasion, while AMPK inhibition promoted trophoblast invasion.
PE complicated placentas are associated with AMPK hyper-activation and consequent alterations in FA oxidation, which inhibit trophoblast invasion.
背景/目的：长期以来，子痫前期（PE）一直被认为是一种胎盘缺血性疾病，尽管关于缺血如何影响胎盘的证据有限。AMP激活的蛋白激酶（AMPK）是细胞能量代谢的关键调节因子，通过增强能量产生在多种缺血性疾病中发挥重要作用。本研究调查了PE患者的胎盘代谢以及AMPK在调节滋养层细胞功能中的作用。
对正常妊娠和合并PE的妊娠胎盘进行气相色谱-质谱联用（GC-MS）以鉴定脂肪酸（FA）代谢指纹图谱，并通过丙二醛（MDA）测量评估总FA氧化。通过定量聚合酶链反应（q-PCR）和蛋白质免疫印迹法评估AMPK-ACC信号通路。将HTR8/SVneo滋养层细胞培养物暴露于不同的氧合条件下以建立体外PE细胞模型；然后通过GC-MS进行代谢物谱分析。在有/无AMPK表达以及操纵AMPK活性后，通过基质胶Transwell试验评估滋养层细胞侵袭，然后通过人绒毛外植体实验进一步验证。
合并PE的妊娠胎盘中，AMPK磷酸化和MDA产生显著升高。PE胎盘顺式双键FA代谢受到抑制，而反式双键FA代谢则被促进。持续低氧的HTR8/SVneo细胞培养条件模拟了AMPK的过度激活，并增强了在PE中观察到的FA氧化。AMPK激活损害滋养层细胞侵袭，而AMPK抑制则促进滋养层细胞侵袭。
合并PE的胎盘与AMPK过度激活以及随之而来的FA氧化改变有关，这会抑制滋养层细胞侵袭。
