Ma Juan, Shen Leilei, Shi Yuan
Department of Neonatology Children's Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing Key Laboratory of Pediatric Metabolism and Inflammatory Diseases, Chongqing, China.
Department of Neonatology, SongShan General Hospital, Chongqing, China.
Front Pharmacol. 2025 Aug 15;16:1621190. doi: 10.3389/fphar.2025.1621190. eCollection 2025.
This study aimed to investigate whether heliox preconditioning (HePC) alleviates neonatal acute respiratory distress syndrome (ARDS) by inhibiting oxidative stress and apoptosis, and to explore its potential mechanism.
Blood samples and bronchoalveolar lavage fluid (BALF) were collected from rat pups were randomly divided into control group, sham group, ARDS group, ARDS + CaMKII group, ARDS + CaMKII group, and ARDS + HePC group. We also investigated the role of CaMKII by manipulating its expression . Inflammatory markers, oxidative stress, apoptosis and activation of signaling pathways were assessed using histological staining, ELISA, Western blotting, qRT-PCR, Ca, immunofluorescence staining, and flow cytometry.
, HePC significantly reduced the expression of CaMKII, inhibited the activation of CaMKII/RyR2, ameliorated the LPS-induced lung histopathological changes in rat pups, reduced lung wet/dry ratios, ROS and MDA levels, and pro-inflammatory cytokine levels, and significantly increased the expression of antioxidant proteins (Nrf2, HO-1 and SOD) and reduced LPS-induced apoptosis. , overexpression of CaMKII increases oxidative stress and activates RyR2, leading to cytoplasmic Ca overload and increased apoptosis. HePC can reverse the above reactions by inhibiting the expression of CaMKII.
HePC may attenuate oxidative stress through CaMKII and alleviate cytoplasmic Ca overload by regulating CaMKII/RyR2, which inhibits apoptosis, exerting lung protection against ARDS.
本研究旨在探讨氦氧混合气预处理（HePC）是否通过抑制氧化应激和细胞凋亡来减轻新生儿急性呼吸窘迫综合征（ARDS），并探索其潜在机制。
从大鼠幼崽中采集血液样本和支气管肺泡灌洗液（BALF），将其随机分为对照组、假手术组、ARDS组、ARDS + CaMKII组、ARDS + CaMKII组和ARDS + HePC组。我们还通过操纵CaMKII的表达来研究其作用。使用组织学染色、酶联免疫吸附测定（ELISA）、蛋白质免疫印迹法、实时定量聚合酶链反应（qRT-PCR）、钙离子测定、免疫荧光染色和流式细胞术评估炎症标志物、氧化应激、细胞凋亡和信号通路的激活情况。
HePC显著降低了CaMKII的表达，抑制了CaMKII/RyR2的激活，改善了脂多糖（LPS）诱导的大鼠幼崽肺组织病理学变化，降低了肺湿/干比、活性氧（ROS）和丙二醛（MDA）水平以及促炎细胞因子水平，并显著增加了抗氧化蛋白（Nrf2、血红素加氧酶-1（HO-1）和超氧化物歧化酶（SOD））的表达，减少了LPS诱导的细胞凋亡。此外，CaMKII的过表达增加了氧化应激并激活了RyR2，导致细胞质钙超载和细胞凋亡增加。HePC可通过抑制CaMKII的表达逆转上述反应。
HePC可能通过CaMKII减轻氧化应激，并通过调节CaMKII/RyR2减轻细胞质钙超载，从而抑制细胞凋亡，对ARDS发挥肺保护作用。
