Guan Enqin, Wang Yue, Wang Caixia, Zhang Ruiyun, Zhao Yiming, Hong Jiang
a Department of Pediatrics , the Affiliated Hospital of Qingdao University , Qingdao , Shandong , China.
b Department of Pediatrics , Qingdao Municipal Hospital , Qingdao , Shandong , China.
Exp Lung Res. 2017 Nov-Dec;43(9-10):378-387. doi: 10.1080/01902148.2017.1384083. Epub 2017 Dec 4.
Receptor-interacting protein (RIP) kinase family members are involved in several biological processes. However, their role in acute lung injury (ALI) is still unclear. In the present study, we aim to determine the expression and function of RIP kinase family in ALI.
In the present study, ALI was induced in BALB/c male mice by intravenously injecting lipopolysaccharide (LPS). The expression levels of the RIP kinase family in ALI mice were determined using western blotting and immunohistochemical staining. The specific RIP-1 inhibitor, necrostatin-1, was used to treat LPS-induced ALI mice, followed by survival time recording, as well as histopathological and immunohistochemical staining of lung tissues, western blotting, myeloperoxidase (MPO) assay and enzyme-linked immunosorbent assay (ELISA) of related cytokines and downstream target expression.
We found that RIP-1 expression was upregulated in the lung of ALI mice and inhibition of RIP-1 by necrostatin-1 significantly prolonged the survival time of mice, which was accompanied by less serve lung injury. Furthermore, lower expression of pro-inflammatory cytokines (interleukin [IL]-6, tumor necrosis factor [TNF]-α, IL-8, cyclooxygenase [COX]-2, monocyte chemoattractant protein [MCP]-1, and IL-1β), MPO and nuclear factor (NF)-κB activation were found in bronchoalveolar lavage fluid (BALF) and lung tissues of necrostatin-1-treated ALI mice. Necrostatin-1 also attenuated LPS-induced pro-inflammatory cytokine expression and NF-κB activation in RAW 264.7 cells.
In summary, necrostatin-1 protected against LPS-induced ALI in mice by inhibiting inflammation and pulmonary NF-κB activation. Thus, necrostatin-1 could be a novel therapeutic strategy for ALI.
受体相互作用蛋白（RIP）激酶家族成员参与多种生物学过程。然而，它们在急性肺损伤（ALI）中的作用仍不清楚。在本研究中，我们旨在确定RIP激酶家族在ALI中的表达及功能。
在本研究中，通过静脉注射脂多糖（LPS）诱导BALB/c雄性小鼠发生ALI。采用蛋白质免疫印迹法和免疫组织化学染色法测定ALI小鼠中RIP激酶家族的表达水平。使用特异性RIP-1抑制剂Necrostatin-1治疗LPS诱导的ALI小鼠，随后记录生存时间，并对肺组织进行组织病理学和免疫组织化学染色、蛋白质免疫印迹分析、髓过氧化物酶（MPO）检测以及相关细胞因子和下游靶点表达的酶联免疫吸附测定（ELISA）。
我们发现ALI小鼠肺组织中RIP-1表达上调，Necrostatin-1抑制RIP-1可显著延长小鼠生存时间，同时减轻肺损伤程度。此外，在Necrostatin-1治疗的ALI小鼠的支气管肺泡灌洗液（BALF）和肺组织中，促炎细胞因子（白细胞介素[IL]-6、肿瘤坏死因子[TNF]-α、IL-8、环氧化酶[COX]-2、单核细胞趋化蛋白[MCP]-1和IL-1β）、MPO的表达降低，核因子（NF）-κB激活减少。Necrostatin-1还可减弱LPS诱导的RAW 264.7细胞中促炎细胞因子的表达和NF-κB激活。
综上所述，Necrostatin-1通过抑制炎症和肺组织中NF-κB激活来保护小鼠免受LPS诱导的ALI。因此，Necrostatin-1可能是一种治疗ALI的新策略。
