Department of Critical Care Medicine, The Affiliated Hospital of Putian University, Putian, China.
School of Wenzhou Medical University, Wenzhou, China.
Int J Immunopathol Pharmacol. 2023 Jan-Dec;37:3946320231192175. doi: 10.1177/03946320231192175.
Acute lung injury (ALI) attracted attention among physicians because of its high mortality. We aimed to determine whether the phosphatidylinositol-3 kinase (PI3K)/protein kinase B (Akt) pathway is involved in the protective effects of penehyclidine hydrochloride (PHC) against lipopolysaccharide (LPS)-induced ALI.
H&E staining was used to observed pathological changes in the lung tissues. ELISA was used to evaluate the concentration of inflammatory mediators in the bronchoalveolar lavage fluid (BALF). White-light microscopy was performed to observe the TUNEL-positive nuclei. The viability of NR8383 alveolar macrophages was determined by using CCK-8. The levels of MPO, MDA, SOD, and GSH-Px were analyzed using ELISA kits. Western blotting was used to evaluate the ERS-associated protein levels and the phosphorylation of PI3K and Akt.
PHC administration defended against LPS-induced histopathological deterioration and increased pulmonary edema and lung injury scores, while all of these beneficial effects were inhibited by LY. In addition, PHC administration mitigated oxidative stress as indicated by decreases in lung myeloperoxidase (MPO) and malondialdehyde (MDA) concentrations, and increases in glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) concentrations. It also alleviated LPS-induced inflammation. PHC administration attenuated apoptosis-associated protein levels, improved cell viability, and decreased the number of TdT-mediated dUTP Nick-End Labeling (TUNEL)-positive cells. Furthermore, PHC inhibited ERS-associated protein levels. Meanwhile, the protection of PHC against inflammation, oxidative stress, apoptosis, and ERS was inhibited by LY. Moreover, PHC administration increased PI3K and Akt phosphorylation, indicating that the upregulation of the PI3K/Akt pathway, while this pathway was inhibited by LY.
PHC significantly activates the PI3K/Akt pathway to ameliorate the extent of damage to pulmonary tissue, inflammation, oxidative stress, apoptosis, and ERS in LPS-induced ALI.
急性肺损伤（ALI）因其高死亡率而引起了医生们的关注。我们旨在确定磷脂酰肌醇-3 激酶（PI3K）/蛋白激酶 B（Akt）通路是否参与了盐酸戊乙奎醚（PHC）对脂多糖（LPS）诱导的 ALI 的保护作用。
通过 H&E 染色观察肺组织的病理变化。通过 ELISA 测定支气管肺泡灌洗液（BALF）中炎症介质的浓度。通过白光显微镜观察 TUNEL 阳性核。用 CCK-8 测定 NR8383 肺泡巨噬细胞的活力。用 ELISA 试剂盒分析 MPO、MDA、SOD 和 GSH-Px 的水平。用 Western blot 评估 ERS 相关蛋白水平和 PI3K 和 Akt 的磷酸化。
PHC 给药可抵抗 LPS 诱导的组织病理学恶化和增加的肺水肿和肺损伤评分，而 LY 抑制了所有这些有益作用。此外，PHC 给药减轻了氧化应激，表现为肺髓过氧化物酶（MPO）和丙二醛（MDA）浓度降低，谷胱甘肽过氧化物酶（GSH-Px）和超氧化物歧化酶（SOD）浓度升高。它还缓解了 LPS 诱导的炎症。PHC 给药可降低凋亡相关蛋白水平，提高细胞活力，并减少 TdT 介导的 dUTP 缺口末端标记（TUNEL）阳性细胞数。此外，PHC 抑制了 ERS 相关蛋白水平。同时，LY 抑制了 PHC 对炎症、氧化应激、凋亡和 ERS 的保护作用。此外，PHC 给药增加了 PI3K 和 Akt 的磷酸化，表明 PI3K/Akt 通路的上调，而 LY 抑制了该通路。
PHC 显著激活 PI3K/Akt 通路，减轻 LPS 诱导的 ALI 中肺组织损伤、炎症、氧化应激、凋亡和 ERS 的程度。
