Department of Thoracic Surgery, Shengjing Hospital of China Medical University, Shenyang, People's Republic of China.
Department of Anesthesiology, Shengjing Hospital of China Medical University, Shenyang, People's Republic of China.
Redox Rep. 2022 Dec;27(1):139-149. doi: 10.1080/13510002.2022.2096339.
Sevoflurane is identified as an effective candidate drug for acute lung injury (ALI) treatment, but its curing effects and detailed mechanisms have not been fully disclosed. The present study was designed to resolve this academic issue.
The ALI mice models were established, and Hematoxylin-eosin staining assay was performed to examine tissue morphologies. Cell viability was determined by CCK-8 assay, and Annexin V-FITC/PI double staining assay was used to examine cell apoptosis. The expression levels of proteins were determined by performing Western Blot analysis and immunofluorescence staining assay. ROS levels were examined by using DCFH-DA staining assay.
In this study, we investigated this issue and the ALI models were respectively established by treating the BALB/c mice and the murine macrophage cell line RAW264.7 with different concentrations of lipopolysaccharide (LPS) in vivo and in vitro, which were subsequently subjected to sevoflurane co-treatment. The results showed that sevoflurane reduced LPS-induced ALI in mice and suppressed LPS-triggered oxidative stress and apoptotic cell death in the RAW264.7 cells. Interestingly, we evidenced that the elimination of reactive oxygen species (ROS) by N-acetyl-L-cysteine (NAC) reversed LPS-induced cell apoptosis in RAW264.7 cells. Then, we verified that sevoflurane suppressed oxidative damages to restrain LPS-induced apoptotic cell death in the RAW264.7 cells through activating the anti-oxidant Keap1/Nrf2 pathway. Mechanistically, sevoflurane down-regulated Keap1 and upregulated Nrf2 in nucleus to activate the downstream anti-oxidant signaling cascades, which further ameliorated LPS-induced cell apoptosis and lung injury by eliminating oxidative damages.
Taken together, our study illustrated that the sevoflurane attenuates LPS-induced ALI by inhibiting oxidative stress-mediated apoptotic cell death and inflammation, and the Keap1/Nrf2 pathway played an important role in this process.
七氟醚被认为是急性肺损伤（ALI）治疗的有效候选药物，但它的治疗效果和详细机制尚未完全揭示。本研究旨在解决这一学术问题。
建立 ALI 小鼠模型，通过苏木精-伊红（Hematoxylin-eosin）染色检测组织形态。通过 CCK-8 检测细胞活力，通过 Annexin V-FITC/PI 双染法检测细胞凋亡。通过 Western blot 分析和免疫荧光染色检测蛋白表达水平。通过 DCFH-DA 染色检测 ROS 水平。
本研究通过体内和体外分别用不同浓度脂多糖（LPS）处理 BALB/c 小鼠和小鼠巨噬细胞 RAW264.7 细胞建立 ALI 模型，然后用七氟醚进行联合处理，研究了这一问题。结果表明，七氟醚减轻了 LPS 诱导的小鼠 ALI，并抑制了 RAW264.7 细胞中 LPS 触发的氧化应激和凋亡性细胞死亡。有趣的是，我们发现 N-乙酰-L-半胱氨酸（NAC）消除活性氧（ROS）可逆转 LPS 诱导的 RAW264.7 细胞凋亡。然后，我们验证了七氟醚通过抑制氧化损伤来抑制 LPS 诱导的 RAW264.7 细胞凋亡性细胞死亡，从而激活抗氧化剂 Kelch 样环氧氯丙烷相关蛋白 1（Keap1）/核因子红细胞 2 相关因子 2（Nrf2）通路。在机制上，七氟醚下调 Keap1 并在上调核内 Nrf2 以激活下游抗氧化信号级联反应，从而通过消除氧化损伤进一步改善 LPS 诱导的细胞凋亡和肺损伤。
综上所述，本研究表明，七氟醚通过抑制氧化应激介导的凋亡性细胞死亡和炎症来减轻 LPS 诱导的 ALI，Keap1/Nrf2 通路在这一过程中发挥重要作用。
