Acetylcholine ameliorated TNF-α-induced primary trophoblast malfunction via muscarinic receptors†

Abstract

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.

Comparative metabolomics analysis of milk components between Italian Mediterranean buffaloes and Chinese Holstein cows based on LC-MS/MS technology

“…The acetylcholine is one of the metabolites synthesized by choline [ 34 ]. Several studies have stated that acetylcholine, acting as a neurotransmitter or an immune modulator, has beneficial effects on the reduction of the oxidative stress, inflammation and apoptosis in a variety of human diseases [ 35 , 36 ]. A recent study also suggested that acetylcholine is required for the acute inflammatory response [ 37 ].…”

“…human diseases [35,36]. A recent study also suggested that acetylcholine is required for the acute inflammatory response [37].…”

The Novel Peptide AEDPPE Alleviates Trophoblast Cell Dysfunction Associated With Preeclampsia by Regulating the NF-κB Signaling Pathway

“…Trophoblast dysfunction is central to the pathogenesis of PE through the action of TNF-α on trophoblast cells, and LPSinjected rats sustain injuries to trophoblast cells; the activation of NF-κB is then the inevitable result of the action of TNF-α and LPS (27)(28)(29)(30)32). It has been demonstrated in several studies that the NF-κB pathway was robustly activated in patients with PE (12,13), and the NF-κB pathway has thus been identified as a potential signaling pathway in the treatment of PE by improving trophoblast function or symptoms in animal models (55).…”

“…The immortalized human first-trimester trophoblast cell-line HTR-8/SVneo cells (ATCC, Rockville, MD, USA) were grown in RPMI 1640 medium (Gibco, Rockville, MD, USA) containing 10% fetal bovine serum (FBS; Gibco, Rockville, MD, USA) in a humid environment containing 5% CO 2 and 95% air at 37 • C. Different concentrations of AEDPPE (1, 5, 10, and 20 µM) were used to stimulate HTR-8/SVneo cells for 24 h. For the treatment of HTR-8/SVneo cells, TNF-α obtained from Sigma-Aldrich (T6674-10UG; St. Louis, MO, USA) was dissolved in double-distilled water, and TNF-α at 20 ng/ml was used to induce inflammation and dysfunction of HTR-8/SVneo cells (27)(28)(29)(30).…”

“…The dysfunction associated with PE in vivo was mimicked by the exploitation of an in vitro TNF-α-injured HTR-8/SVneo cell model (27)(28)(29)(30), and the effects of AEDPPE on cellular proliferation and apoptosis were explored using CCK-8 assays and flow cytometry. We observed no effect on the proliferation or apoptosis of HTR-8/SVneo cells after TNF-α treatment with or without AEDPPE compared to the control group (Figures 3A,B).…”