Department of Interventional Radiology, Renmin Hospital of Wuhan University, Wuhan, Hubei, China (mainland).
Department of Thoracic Surgery, Renmin Hospital of Wuhan University, Wuhan, Hubei, China (mainland).
Med Sci Monit. 2021 Apr 3;27:e930429. doi: 10.12659/MSM.930429.
BACKGROUND Acute lung injury (ALI) results from damage to the alveolar capillary endothelial cells and can result in acute respiratory distress syndrome (ARDS). This study aimed to investigate murine lung vascular endothelial cells (MLECs) damage in a murine model of lipopolysaccharide (LPS)-induced ALI. MATERIAL AND METHODS Mice were injected with LPS to induce an acute lung injury model. An adenovirus transfection system was used to overexpress or knockdown DUSP12 in mice. MLECs were isolated, cultured and transfected with DUSP12-overexpressing adenovirus or with DUSP12 siRNA to knockdown DUSP12. LPS was used to establish a cell injury model. ELISA and RT-PCR were used to examine cell inflammation. LPS-induced oxidative stress was also evaluated using commercial kits. RESULTS A decreased level of DUSP12 was observed in MLECs treated with LPS. DUSP12 overexpression in mice attenuated LPS-induced lung inflammation and lung injury, as reflected by reduced levels of proinflammatory cytokines. Mice with DUSP12 knockdown exhibited worsened lung inflammation and injury. In vitro, DUSP12 overexpression in endothelial cells ameliorated LPS-induced inflammation, apoptosis, and oxidative stress. DUSP12 silencing in endothelial cells aggravated LPS-induced inflammation, apoptosis, and oxidative stress. Furthermore, we found that DUSP12 directly bound to apoptosis signal-regulating kinase 1 (ASK1) to inhibit Jun N-terminal kinase activation (JNK). A JNK1/2 inhibitor and ASK1 siRNA ameliorated the exacerbating effects of DUSP12 knockdown in vitro. CONCLUSIONS Our data demonstrated that DUSP12 suppressed MLEC injury in response to LPS insult by regulating the ASK1/JNK pathway.
急性肺损伤 (ALI) 是由肺泡毛细血管内皮细胞损伤引起的，并可能导致急性呼吸窘迫综合征 (ARDS)。本研究旨在探讨脂多糖 (LPS) 诱导的 ALI 小鼠模型中肺血管内皮细胞 (MLEC) 的损伤。
用 LPS 注射诱导小鼠急性肺损伤模型。采用腺病毒转染系统过表达或敲低 DUSP12。分离、培养 MLEC 并转染 DUSP12 过表达腺病毒或 DUSP12 siRNA 以敲低 DUSP12。用 LPS 建立细胞损伤模型。ELISA 和 RT-PCR 用于检测细胞炎症。使用商业试剂盒评估 LPS 诱导的氧化应激。
用 LPS 处理的 MLEC 中观察到 DUSP12 水平降低。DUSP12 在小鼠中的过表达减轻了 LPS 诱导的肺炎症和肺损伤，反映在促炎细胞因子水平降低。DUSP12 敲低的小鼠表现出更严重的肺炎症和损伤。在体外，内皮细胞中 DUSP12 的过表达改善了 LPS 诱导的炎症、凋亡和氧化应激。内皮细胞中 DUSP12 的沉默加剧了 LPS 诱导的炎症、凋亡和氧化应激。此外，我们发现 DUSP12 直接与凋亡信号调节激酶 1 (ASK1) 结合以抑制 Jun N-末端激酶激活 (JNK)。JNK1/2 抑制剂和 ASK1 siRNA 改善了体外 DUSP12 敲低的加剧作用。
我们的数据表明，DUSP12 通过调节 ASK1/JNK 通路抑制 LPS 损伤诱导的 MLEC 损伤。
