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.

中文翻译：

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乙酰胆碱通过毒蕈碱受体改善 TNF-α 诱导的原发性滋养细胞功能障碍。

滋养层细胞的氧化应激和凋亡与先兆子痫 (PE) 相关。大量研究表明，乙酰胆碱 (ACh) 是主要的迷走神经递质，在减轻各种人类疾病的氧化应激、炎症和细胞凋亡方面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 处理的人原代滋养层细胞的影响。进行蛋白质印迹、CCK-8、DHE、TUNEL 免疫荧光染色、transwell 测定和伤口愈合测定以评估 ACh 在体外的作用。我们发现先兆子痫妇女胎盘的 TNF-α 表达和凋亡指数均高于正常胎盘。TNF-α 增强氧化应激并增加 TUNEL 阳性细胞核的数量、Bax/Bcl-2 比值、和切割的 caspase-3/caspase-3 比率，同时降低原代人滋养层细胞的细胞活力。TNF-α 促进细胞迁移和侵袭。PDTC 是一种选择性 NF-κB 抑制剂，可显着减弱 TNF-α 诱导的作用。ACh 处理减弱了氧化应激和细胞凋亡，同时进一步促进了 TNF-α 处理的原代滋养层细胞的迁移和侵袭。ACh 的作用可以被毒蕈碱受体拮抗剂阿托品逆转。总体而言，我们的研究结果表明，ACh 通过毒蕈碱受体显着改善 TNF-α 诱导的人原代滋养层细胞的氧化应激和凋亡。这是第一次将迷走神经活动的改善作为 PE 样滋养层细胞的治疗策略，表明其在临床实践中的潜在价值。