Department of Pharmacology, School of Medicine, Xi'an Jiaotong University, Xi'an, China.
Department of Pharmacy, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China.
Biol Reprod. 2020 Dec 1;103(6):1238-1248. doi: 10.1093/biolre/ioaa158.
Oxidative stress and apoptosis of trophoblasts are involved in preeclampsia (PE). Numerous studies have shown that acetylcholine (ACh), the principal vagal neurotransmitter, plays a crucial role in attenuating oxidative stress, inflammation, and apoptosis in a variety of human diseases. However, the role of ACh in PE management remains unclear. Here, we aimed to determine the effects of ACh on TNF-α-treated human primary trophoblast cells. Western blotting, CCK-8, DHE, TUNEL immunofluorescence staining, transwell assays, and wound-healing assays were performed to evaluate the role of ACh in vitro. We found that both TNF-α expression and the apoptotic index were higher in placentas from preeclamptic women than in normal placentas. TNF-α enhanced oxidative stress and increased the number of TUNEL-positive nuclei, Bax/Bcl-2 ratio, and the cleaved caspase-3/caspase-3 ratio while decreasing cell viability in primary human trophoblast cells. TNF-α promoted cell migration and invasion. PDTC, a selective NF-κB inhibitor, significantly blunted TNF-α-induced effects. ACh treatment attenuated oxidative stress and apoptosis while further promoting migration and invasion of TNF-α-treated primary trophoblast cells. The effects of ACh could be reversed by the muscarinic receptor antagonist atropine. Overall, our findings indicate that ACh significantly ameliorates TNF-α-induced oxidative stress and apoptosis of human primary trophoblast cells via muscarinic receptors. This is the first time that the improvement of vagal activity served as a therapeutic strategy for PE-like trophoblasts, suggesting its potential value in clinical practice.
氧化应激和滋养细胞凋亡参与了子痫前期（PE）的发生。大量研究表明，乙酰胆碱（ACh）作为主要的迷走神经递质，在多种人类疾病中对减轻氧化应激、炎症和细胞凋亡起着至关重要的作用。然而，ACh 在 PE 管理中的作用仍不清楚。在这里，我们旨在确定 ACh 对 TNF-α 处理的人原代滋养细胞的影响。采用 Western blot、CCK-8、DHE、TUNEL 免疫荧光染色、Transwell 检测和划痕愈合检测等方法在体外评估 ACh 的作用。我们发现，子痫前期患者胎盘组织中 TNF-α 的表达和凋亡指数均高于正常胎盘组织。TNF-α增强了氧化应激，增加了 TUNEL 阳性核、Bax/Bcl-2 比值和 cleaved caspase-3/caspase-3 比值，同时降低了原代人滋养细胞的活力。TNF-α促进了细胞迁移和侵袭。NF-κB 选择性抑制剂 PDTC 显著减弱了 TNF-α 诱导的作用。ACh 处理减轻了氧化应激和凋亡，同时进一步促进了 TNF-α 处理的原代滋养细胞的迁移和侵袭。毒蕈碱受体拮抗剂阿托品可逆转 ACh 的作用。总的来说，我们的研究结果表明，ACh 通过毒蕈碱受体显著改善了 TNF-α 诱导的人原代滋养细胞的氧化应激和凋亡。这是首次报道迷走神经活性的改善作为子痫前期样滋养细胞的治疗策略，提示其在临床实践中的潜在价值。
