Pyridostigmine inhibits placental necroptosis and ameliorates preeclampsia-like symptoms in rats

Necroptosis contributes to the pathogenesis of preeclampsia (PE). However, effective therapeutic strategies targeting placental necroptosis remain limited. The significance of the pharmacologically enhanced non-neuronal cholinergic signaling by pyridostigmine (PYR) on placental necroptosis was investigated. The expression of necroptosis-related markers was assessed in placentas from PE women and rats with reduced uterine perfusion pressure (RUPP). RUPP rats were treated with PYR, in presence or absence of the α7 nicotinic acetylcholine receptor (α7nAChR) antagonist, the α-bungarotoxin (α-BGT), or the necroptosis inhibitor necrostatin-1 (Nec-1) to evaluate the therapeutic potential of targeting necroptosis in response to placental ischemia. At cellular level, the effects of acetylcholine (ACh) on hypoxic trophoblast cells were assessed in vitro. The placental expression of receptor-interacting protein kinase 1 (RIPK1), phosphorylated RIPK1, mixed lineage kinase domain-like protein (MLKL), and phosphorylated MLKL and the maternal blood pressure were elevated in PE women and RUPP rats. Nec-1 and PYR reversed these changes, and attenuated both oxidative stress and inflammation in RUPP rats. α-BGT abolished the effects of PYR. Moreover, ACh suppressed necroptosis and inflammatory responses and restored trophoblast migratory capacity under hypoxic conditions. This study revealed the potential of targeting necroptosis in placental ischemia-driven PE model. PYR alleviated PE features and placental necroptosis and inflammation potentially via activation of α7nAChR, which highlight the non-neuronal cholinergic system as a promising therapeutic target for PE.