Department of Obstetrics & Gynecology, University of California, Irvine, Irvine, California, USA.
Biol Reprod. 2017 Sep 1;97(3):478-489. doi: 10.1093/biolre/iox105.
Endogenous hydrogen sulfide (H2S), mainly synthesized by cystathionine β-synthase (CBS) and cystathionine γ-lyase (CTH), has been implicated in regulating placental angiogenesis; however, the underlying mechanisms are unknown. This study was to test a hypothesis that trophoblasts synthesize H2S to promote placental angiogenesis. Human choriocarcinoma-derived BeWo cells expressed both CBS and CTH proteins, while the first trimester villous trophoblast-originated HTR-8/SVneo cells expressed CTH protein only. The H2S producing ability of BeWo cells was significantly inhibited by either inhibitors of CBS (carboxymethyl hydroxylamine hemihydrochloride, CHH) or CTH (β-cyano-L-alanine, BCA) and that in HTR-8/SVneo cells was inhibited by CHH only. H2S donors stimulated cell proliferation, migration, and tube formation in ovine placental artery endothelial cells (oFPAECs) as effectively as vascular endothelial growth factor. Co-culture with BeWo and HTR-8/SVneo cells stimulated oFPAEC migration, which was inhibited by CHH or BCA in BeWo but CHH only in HTR-8/SVneo cells. Primary human villous trophoblasts (HVT) were more potent than trophoblast cell lines in stimulating oFPAEC migration that was inhibited by CHH and CHH/BCA combination in accordance with its H2S synthesizing activity linked to CBS and CTH expression patterns. H2S donors activated endothelial nitric oxide synthase (NOS3), v-AKT murine thymoma viral oncogene homolog 1 (AKT1), and extracellular signal-activated kinase 1/2 (mitogen-activated protein kinase 3/1, MAPK3/1) in oFPAECs. H2S donor-induced NOS3 activation was blocked by AKT1 but not MAPK3/1 inhibition. In keeping with our previous studies showing a crucial role of AKT1, MAPK3/1, and NOS3/NO in placental angiogenesis, these data show that trophoblast-derived endogenous H2S stimulates placental angiogenesis, involving activation of AKT1, NOS3/NO, and MAPK3/1.
内源性硫化氢 (H2S) 主要由半胱氨酸 β-合酶 (CBS) 和半胱氨酸 γ-裂解酶 (CTH) 合成，被认为参与调节胎盘血管生成；然而，其潜在机制尚不清楚。本研究旨在验证一个假设，即滋养细胞合成 H2S 以促进胎盘血管生成。人绒毛膜癌细胞系 BeWo 表达 CBS 和 CTH 蛋白，而来源于早期妊娠绒毛滋养层的 HTR-8/SVneo 细胞仅表达 CTH 蛋白。BeWo 细胞的 H2S 产生能力被 CBS 抑制剂（羟甲基半胱氨酸盐酸盐，CHH）或 CTH 抑制剂（β-氰基-L-丙氨酸，BCA）显著抑制，而 HTR-8/SVneo 细胞的 H2S 产生能力仅被 CHH 抑制。H2S 供体刺激绵羊胎盘动脉内皮细胞（oFPAECs）的细胞增殖、迁移和管形成，与血管内皮生长因子的效果相当。BeWo 和 HTR-8/SVneo 细胞共培养可刺激 oFPAEC 迁移，而 CHH 或 BCA 在 BeWo 细胞中抑制该迁移，而仅在 HTR-8/SVneo 细胞中 CHH 抑制该迁移。与胎盘滋养层细胞系相比，原代人绒毛滋养层（HVT）更能刺激 oFPAEC 迁移，而 CHH 和 CHH/BCA 联合抑制其 H2S 合成活性，与 CBS 和 CTH 表达模式相关。H2S 供体激活内皮型一氧化氮合酶（NOS3）、v-AKT 鼠胸腺瘤病毒癌基因同源物 1（AKT1）和细胞外信号调节激酶 1/2（丝裂原激活蛋白激酶 3/1，MAPK3/1）在 oFPAECs 中。H2S 供体诱导的 NOS3 激活被 AKT1 而非 MAPK3/1 抑制所阻断。与我们之前的研究表明 AKT1、MAPK3/1 和 NOS3/NO 在胎盘血管生成中的关键作用一致，这些数据表明，滋养细胞衍生的内源性 H2S 刺激胎盘血管生成，涉及 AKT1、NOS3/NO 和 MAPK3/1 的激活。
