Department of Critical Care Medicine, The Affiliated Hospital of Putian University, Putian 351100, China.
Department of Respiratory and Critical Care, The Affiliated Hospital of Putian University, Putian 351100, China.
Gene. 2019 Dec 30;721:144095. doi: 10.1016/j.gene.2019.144095. Epub 2019 Aug 30.
Penehyclidine hydrochloride (PHC) is a novel anticholinergic drug applied broadly in surgeries as a preanesthetic medication. A substantial amount of research indicates that PHC has lung defensive properties. Considering that endoplasmic reticulum (ER) stress exerts a crucial function in cell apoptosis associated with the lipopolysaccharides (LPS)-induced acute lung injury (ALI) model, we aimed to determine whether regulation of ER stress in the LPS-induced ALI model was associated with the lung defensive role of PHC. Adult male SD rats were administered LPS (5 mg/kg, intratracheally) followed by PHC (1.0 mg/kg, intravenously) for 24 h. The NR8383 alveolar macrophages were randomly separated into Sham, LPS (100 ng/mL), and PHC (1, 2.5, or 5 μg/mL) + LPS groups. PHC (1, 2.5, or 5 μg/mL) + LPS groups were treated with PHC alone for 1 h after LPS exposure. Posttreatment with PHC relieved LPS-induced pulmonary impairment and blocked LPS-mediated lung apoptosis, indicated by the downregulation of the lung apoptotic indicators malondialdehyde and superoxide dismutase in serum at 24 h after LPS-induced ALI. PHC (1-5 μg/mL) did not influence the activity of cultivated NR8383 alveolar macrophages in vitro. However, postconditioning with PHC dosage-dependently reduced LPS-mediated cell apoptosis. Additionally, many studies have indicated that PHC administration inhibits ER stress and initiates hairy and enhancer of split 1 (Hes1)/(Notch1) signaling by decreasing phosphorylated α subunit of eukaryotic initiation factor 2α (p-eIF2α)/eukaryotic translation initiation factor 2α (eIF2α) and Phospho-protein kinase R-like ER kinase (p-PERK)/ protein kinase R-like ER kinase (PERK) proportions; inhibiting C/EBP-homologous protein (CHOP), activating transcription factor 4 (ATF4), caspase-3, and Bcl2-associated x (Bax) activity; and enhancing notch1 intracellular domain (NICD), Notch1, B-cell lymphoma-2 (Bcl-2), and Hes1 activity in vivo and in vitro. In addition, the defensive functions of PHC on LPS-activated NR8383 alveolar macrophages were abrogated through the Notch1 pathway antagonist [(3,5-difluorophenacetyl)-1-alanyl] -phenylglycine-butyl ester (DAPT). In conclusion, PHC alleviates LPS-induced ALI by ameliorating ER stress-mediated apoptosis and promoting Hes1/Notch1 signaling in vivo and in vitro.
盐酸戊乙奎醚（PHC）是一种新型的抗胆碱能药物，广泛应用于手术中作为麻醉前用药。大量研究表明，PHC 具有肺保护作用。鉴于内质网（ER）应激在脂多糖（LPS）诱导的急性肺损伤（ALI）模型中与细胞凋亡有关，我们旨在确定 LPS 诱导的 ALI 模型中 ER 应激的调节是否与 PHC 的肺保护作用有关。成年雄性 SD 大鼠给予 LPS（5mg/kg，气管内），然后给予 PHC（1.0mg/kg，静脉内）24h。NR8383 肺泡巨噬细胞随机分为 Sham、LPS（100ng/mL）和 PHC（1、2.5 或 5μg/mL）+LPS 组。PHC（1、2.5 或 5μg/mL）+LPS 组在 LPS 暴露后单独用 PHC 处理 1h。用 PHC 处理可减轻 LPS 诱导的肺损伤，并阻断 LPS 介导的肺细胞凋亡，这表现在 LPS 诱导的 ALI 后 24h 血清中肺凋亡指标丙二醛和超氧化物歧化酶的下调。PHC（1-5μg/mL）不影响体外培养的 NR8383 肺泡巨噬细胞的活性。然而，用 PHC 后处理可剂量依赖性地减少 LPS 介导的细胞凋亡。此外，许多研究表明，PHC 给药通过降低磷酸化真核起始因子 2α（p-eIF2α）/真核翻译起始因子 2α（eIF2α）和磷酸蛋白激酶 R 样内质网激酶（p-PERK）/蛋白激酶 R 样内质网激酶（PERK）比例，抑制内质网应激并启动 hairy 和 enhancer of split 1（Hes1）/Notch1 信号通路，从而抑制 C/EBP 同源蛋白（CHOP）、激活转录因子 4（ATF4）、半胱天冬酶-3 和 Bcl2 相关 X（Bax）的活性，并增强 notch1 细胞内结构域（NICD）、Notch1、B 细胞淋巴瘤-2（Bcl-2）和 Hes1 的活性，在体内和体外。此外，通过 Notch1 通路拮抗剂 [(3,5-二氟苯乙酰基)-1-丙氨酰基]-苯丙氨酸-丁酯（DAPT），PHC 对 LPS 激活的 NR8383 肺泡巨噬细胞的防御功能被阻断。总之，PHC 通过改善 ER 应激介导的凋亡并促进体内和体外的 Hes1/Notch1 信号通路，减轻 LPS 诱导的 ALI。
