Zhang Hong-Hai, Chen Jennifer C, Sheibani Lili, Lechuga Thomas J, Chen Dong-Bao
Department of Obstetrics & Gynecology, University of California, Irvine, California 92697.
J Clin Endocrinol Metab. 2017 Jul 1;102(7):2382-2393. doi: 10.1210/jc.2017-00437.
Augmented uterine artery (UA) production of vasodilators, including nitric oxide (NO) and hydrogen sulfide (H2S), has been implicated in pregnancy-associated and agonist-stimulated rise in uterine blood flow that is rate-limiting to pregnancy health.
Developing a human UA endothelial cell (hUAEC) culture model from main UAs of nonpregnant (NP) and pregnant (P) women for testing a hypothesis that pregnancy augments endothelial NO and H2S production and endothelial reactivity to vascular endothelial growth factor (VEGF).
Main UAs from NP and P women were used for developing hUAEC culture models. Comparisons were made between NP- and P-hUAECs in in vitro angiogenesis, activation of cell signaling, expression of endothelial NO synthase (eNOS) and H2S-producing enzymes cystathionine β-synthase (CBS) and cystathionine γ-lyase, and NO/H2S production upon VEGF stimulation.
NP- and P-hUAECs displayed a typical cobblestone-like shape in culture and acetylated low-density lipoprotein uptake, stained positively for endothelial and negatively for smooth muscle markers, maintained key signaling proteins during passage, and had statistically significant greater eNOS and CBS proteins in P- vs NP-hUAECs. Treatment with VEGF stimulated in vitro angiogenesis and eNOS protein and NO production only in P-hUEACs and more robust cell signaling in P- vs NP-hUAECs. VEGF stimulated CBS protein expression, accounting for VEGF-stimulated H2S production in hUAECs.
Comparisons between NP- and P-hUAECs reveal that pregnancy augments VEGF-stimulated in vitro angiogenesis and NO/H2S production in hUAECs, showing that the newly established hUAEC model provides a critical in vitro tool for understanding human uterine hemodynamics.
子宫动脉（UA）产生的血管舒张剂，包括一氧化氮（NO）和硫化氢（H2S），与妊娠相关以及激动剂刺激引起的子宫血流量增加有关，而子宫血流量增加对妊娠健康具有限速作用。
从非妊娠（NP）和妊娠（P）女性的主要子宫动脉中建立人子宫动脉内皮细胞（hUAEC）培养模型，以检验妊娠会增强内皮细胞产生NO和H2S以及内皮细胞对血管内皮生长因子（VEGF）反应性这一假设。
使用NP和P女性的主要子宫动脉建立hUAEC培养模型。对NP-hUAEC和P-hUAEC在体外血管生成、细胞信号激活、内皮型一氧化氮合酶（eNOS）和产生H2S的酶胱硫醚β-合酶（CBS）及胱硫醚γ-裂解酶的表达，以及VEGF刺激后的NO/H2S产生情况进行比较。
NP-hUAEC和P-hUAEC在培养中呈现典型的鹅卵石样形态，具有乙酰化低密度脂蛋白摄取能力，内皮标志物染色呈阳性，平滑肌标志物染色呈阴性，传代过程中维持关键信号蛋白，且P-hUAEC中的eNOS和CBS蛋白在统计学上显著高于NP-hUAEC。VEGF处理仅在P-hUEAC中刺激体外血管生成、eNOS蛋白表达和NO产生，且P-hUAEC中的细胞信号比NP-hUAEC更强烈。VEGF刺激CBS蛋白表达，这解释了hUAEC中VEGF刺激产生H2S的现象。
NP-hUAEC和P-hUAEC的比较表明，妊娠增强了VEGF刺激的hUAEC体外血管生成和NO/H2S产生，表明新建立的hUAEC模型为理解人类子宫血流动力学提供了关键的体外工具。
