Li Hong-Xia, Liang Xiao-Yan, Wu Jiong-He, Yuan Ya-Ping, Gao Yue, Cai Shao-Hua
Department of Respiratory and Critical Care Medicine, The Second Medical Center, National Clinical Research Center for Geriatric Diseases, Chinese PLA General Hospital, Beijing 100853, China.
Department of Respiratory Medicine in Special Wards, Chinese PLA General Hospital, Beijing 100853, China.
Toxicol Appl Pharmacol. 2021 Jul 1;422:115460. doi: 10.1016/j.taap.2021.115460. Epub 2021 Mar 24.
To explore the protective mechanism of simvastatin in acute lung injury (ALI), the lipopolysaccharide (LPS) induced (5 mg/kg) ALI rat model was used to examine the effects of simvastatin. Following simvastatin treatment, the histopathological evaluation of lung tissues was made using hematoxylin and eosin (H&E) staining. Also, myeloperoxidase (MPO) activity and the levels of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and IL-10 were determined by ELISA. Blood gas analyses of arterial blood samples were performed to assess the pulmonary gas exchange. Moreover, the neutrophil count and total protein content were determined in the bronchoalveolar lavage (BAL) fluid. The ratio of wet lung to dry lung (W/D) and the alveolar fluid clearance (AFC) were calculated to estimate the severity of edema. Lastly, the levels of A2BAR, CFTR, claudin4, and claudin18 were also measured by qRT-PCR and Western blotting. Simvastatin treatment, in a dose-related manner, markedly improved the lung histological injury and decreased the levels of TNF-α, IL-1β, and increased IL-10 in LPS induced ALI. Also, pulmonary neutrophil count was alleviated. Besides, a decreased ratio of W/D lung also confirmed the simvastatin intervention. Notably, simvastatin reduced the levels of A2BAR, CFTR, and claudin18 but upregulated claudin4 in lung tissues. Additionally, treatment with PSB1115, an antagonist of A2BAR, countered the protective effect of simvastatin in ALI. Our study demonstrates that simvastatin has a protective effect against LPS-induced ALI by activating A2BAR and should be exploited as a novel therapeutic target for the treatment of ALI.
为探究辛伐他汀在急性肺损伤（ALI）中的保护机制，采用脂多糖（LPS）诱导（5 mg/kg）的ALI大鼠模型来检测辛伐他汀的作用。给予辛伐他汀治疗后，使用苏木精-伊红（H&E）染色对肺组织进行组织病理学评估。此外，通过酶联免疫吸附测定（ELISA）法测定髓过氧化物酶（MPO）活性以及肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和IL-10的水平。对动脉血样本进行血气分析以评估肺气体交换情况。此外，测定支气管肺泡灌洗（BAL）液中的中性粒细胞计数和总蛋白含量。计算肺湿重与干重之比（W/D）和肺泡液体清除率（AFC）以评估水肿的严重程度。最后，通过定量逆转录聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法（Western blotting）测定A2B受体（A2BAR）、囊性纤维化跨膜传导调节因子（CFTR）、紧密连接蛋白4（claudin4）和紧密连接蛋白18（claudin18）的水平。辛伐他汀治疗以剂量相关的方式显著改善了肺组织学损伤，降低了LPS诱导的ALI中TNF-α、IL-1β的水平，并增加了IL-10的水平。此外，肺中性粒细胞计数也有所减轻。此外，肺W/D比值降低也证实了辛伐他汀的干预作用。值得注意的是，辛伐他汀降低了肺组织中A2BAR、CFTR和claudin18的水平，但上调了claudin4的水平。此外，使用A2BAR拮抗剂PSB1115进行治疗可抵消辛伐他汀在ALI中的保护作用。我们的研究表明，辛伐他汀通过激活A2BAR对LPS诱导的ALI具有保护作用，应将其开发为治疗ALI的新型治疗靶点。
