Zhou Mi, Meng Ling, He Qinke, Ren Chunguang, Li Changyi
Department of Respiratory and Critical Care, Second Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Laboratory of Developmental Biology, Department of Cell Biology and Genetics, School of Basic Medical Sciences, Chongqing Medical University, Chongqing, China.
Front Pharmacol. 2024 Jan 15;15:1321095. doi: 10.3389/fphar.2024.1321095. eCollection 2024.
Acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) is a common respiratory disease characterized by persistent hypoxemia and an uncontrolled inflammatory response. Valsartan, an angiotensin II type 1 receptor antagonist, is clinically used to treat hypertension and has anti-inflammatory and antioxidant effects on gefitinib-induced pneumonia in rats. However, the potential therapeutic effects of valsartan on lipopolysaccharide (LPS)-induced ALI remain unclear. This study investigated the protective role of valsartan in LPS-induced ALI and its underlying mechanisms. LPS-treated BEAS-2B cells and ALI mouse model were established. BEAS-2B cells were treated with LPS (10 μg/mL) for 24h, with or without valsartan (20, 40, and 80 µM). For ALI mouse models, LPS (5 mg/kg) was administered through intratracheal injection to treat the mice for 24h, and valsartan (10 or 30 mg/kg) was injected intraperitoneally twice 2 h before and 12 h after the LPS injection. Pulmonary functional parameters were examined by an EMKA pulmonary system. Hematoxylin and eosin staining, flow cytometry, CCK-8 assay, qRT-PCR, ELISA, immunofluorescence, Western blotting, and related commercial kits were used to assess the pathological damage to the lungs, neutrophil recruitment in the lung tissue and bronchoalveolar lavage fluid (BALF), cell viability, inflammation, oxidative activity, and mucus production, respectively. Potential mechanisms were further explored using network pharmacology and Western blotting. Valsartan rescued LPS-reduced cell viability of BEAS-2B cells, improved the pulmonary function, ameliorated pathological lung injury in mice with ALI, ameliorated LPS-induced neutrophil recruitment in BALF and lung tissue of mice, attenuated oxidative stress by increasing the level of SOD and decreasing that of MDA and GSSG, inhibited LPS-induced MUC5AC overproduction, decreased the LPS-induced increase in expression of pro-inflammatory cytokines/chemokines including TNF-α, IL-6, IL-1β, CXCL-1 and CXCL-2, and restored the expression of anti-inflammatory IL-10. Mechanistic studies showed that valsartan inhibits LPS-induced phosphorylation of nuclear factor-kappa B (NF-κΒ) and mitogen-activated protein kinases (MAPKs) including P38, extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK) in both LPS-treated cells and the mouse model of ALI. Valsartan protects against LPS-induced ALI by attenuating oxidative stress, reducing MUC5AC production, and attenuating the inflammatory response that may involve MAPK and NF-κΒ pathways.
急性肺损伤（ALI）/急性呼吸窘迫综合征（ARDS）是一种常见的呼吸系统疾病，其特征为持续性低氧血症和失控的炎症反应。缬沙坦是一种血管紧张素II 1型受体拮抗剂，临床上用于治疗高血压，对吉非替尼诱导的大鼠肺炎具有抗炎和抗氧化作用。然而，缬沙坦对脂多糖（LPS）诱导的ALI的潜在治疗作用仍不清楚。本研究调查了缬沙坦在LPS诱导的ALI中的保护作用及其潜在机制。建立了LPS处理的BEAS-2B细胞和ALI小鼠模型。BEAS-2B细胞用LPS（10μg/mL）处理24小时，分别添加或不添加缬沙坦（20、40和80μM）。对于ALI小鼠模型，通过气管内注射给予LPS（5mg/kg）处理小鼠24小时，在LPS注射前2小时和注射后12小时腹腔注射两次缬沙坦（10或30mg/kg）。通过EMKA肺功能系统检测肺功能参数。分别使用苏木精和伊红染色、流式细胞术、CCK-8测定、qRT-PCR、ELISA、免疫荧光、蛋白质印迹以及相关商业试剂盒评估肺组织的病理损伤、肺组织和支气管肺泡灌洗液（BALF）中的中性粒细胞募集、细胞活力、炎症、氧化活性和黏液分泌。使用网络药理学和蛋白质印迹进一步探索潜在机制。缬沙坦挽救了LPS降低的BEAS-2B细胞活力，改善了肺功能，减轻了ALI小鼠的肺病理损伤，减轻了LPS诱导的小鼠BALF和肺组织中的中性粒细胞募集，通过提高超氧化物歧化酶（SOD）水平并降低丙二醛（MDA）和谷胱甘肽二硫化物（GSSG）水平减轻了氧化应激，抑制了LPS诱导的MUC5AC过度产生，降低了LPS诱导的促炎细胞因子/趋化因子（包括肿瘤坏死因子-α、白细胞介素-6、白细胞介素-1β、CXC趋化因子配体1（CXCL-1）和CXC趋化因子配体2（CXCL-2））表达的增加，并恢复了抗炎性白细胞介素-10的表达。机制研究表明，缬沙坦在LPS处理的细胞和ALI小鼠模型中均抑制LPS诱导的核因子-κB（NF-κB）和丝裂原活化蛋白激酶（MAPK）（包括P38、细胞外信号调节激酶（ERK）和c-Jun氨基末端激酶（JNK））的磷酸化。缬沙坦通过减轻氧化应激、减少MUC5AC产生以及减轻可能涉及MAPK和NF-κB途径的炎症反应来预防LPS诱导的ALI。
