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Activation of necroptosis in a rat model of acute respiratory distress syndrome induced by oleic acid

PAN Long, YAO Dun-Chen, YU Yu-Zhong, CHEN Bing-Jun, LI Sheng-Jie, HU Gui-He, XI Chang, WANG Zi-Hui, LI Jian-Hua, LONG Jie, TU Yong-Sheng

Department of Physiology, School of Basic Sciences； The Third Clinical Medical College； The First Clinical Medical College； The Second Clinical Medical College； 5Department of Pathology, School of Basic Sciences, Guangzhou Medical University, Guangzhou 511436, China

Abstract

The present study was aimed to investigate the role of necroptosis in the pathogenesis of acute respiratory distress syndrome (ARDS). The rat model of ARDS was induced by intravenous injection of oleic acid (OA), and observed for 4 h. The lung injury was evaluated by arterial blood gas, lung wet-dry weight ratio (W/D) and histological analyses. Simultaneously, bronchoalveolar lavage fluid (BALF) was collected for total and differential cell analysis and total protein determination. Tumor necrosis factor alpha (TNF-α) level in BALF was determined with a rat TNF-α ELISA kit. Expressions of receptor interacting protein kinase 1 (RIPK1), RIPK3 and mixed lineage kinase domain-like protein (MLKL) in lung tissue were determined by Western blot and immunohistochemical staining. The interaction between RIPK1 and RIPK3 was explored by immunoprecipitation. The results showed that, compared with those in control group, total white blood cells count (WBC), polymorphonuclear percentage (PMN%), total protein concentration, TNF-α level in BALF, W/D, and the alveolar-arterial oxygen tension difference (P(A-a)O2) in OA group were significantly increased at 4 h after OA injection. Western blot and immunostaining further showed remarkably increased expressions of RIPK1, RIPK3 and MLKL in lung tissue from OA group. Additionally, immunoprecipitation results indicated an enforced interaction between RIPK1 and RIPK3 in OA group. Collectively, the TNF-α level in BALF and the RIPK1-RIPK3-MLKL signaling pathway in lung tissue were found to be upregulated and activated with the process of ARDS. These findings implicate that RIPK1/RIPK3-mediated necroptosis plays a possible role in the pathogenesis of ARDS, which may provide a new idea to develop novel drugs for the therapy of ARDS.

Key words: Acute respiratory distress syndrome; necroptosis; TNF-α; receptor interacting protein kinase 1 (RIPK1); RIPK3; mixed lineage kinase domain-like protein

Received: 2016-04-28　　Accepted: 2016-07-22

Corresponding author: 涂永生　　E-mail: tuys@gzhmu.edu.cn

Citing This Article：

PAN Long, YAO Dun-Chen, YU Yu-Zhong, CHEN Bing-Jun, LI Sheng-Jie, HU Gui-He, XI Chang, WANG Zi-Hui, LI Jian-Hua, LONG Jie, TU Yong-Sheng. Activation of necroptosis in a rat model of acute respiratory distress syndrome induced by oleic acid. Acta Physiol Sin 2016; 68 (5): 661-668 (in Chinese with English abstract).