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Norepinephrine induces apoptosis in neonatal rat endothelial cells via a ROS-dependent JNK activation pathway

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Abstract

Our previous study demonstrated that norepinephrine (NE) induces endothelial apoptosis mainly through down-regulation of Bcl-2 protein and activation of the β-adrenergic and caspase-2 pathways. However, whether reactive oxygen species (ROS) and mitogen-activated protein kinases (MAPKs) are involved in this signal transduction remains unknown. Endothelial cells cultured from neonatal rat heart were treated with 100 μM NE. Proteins of MAPKs and Bcl-2 family were assayed by Western blotting. Apoptosis was determined by terminal deoxynucleotidyl transferase-mediated nick end-labeling assay. ROS was analyzed with flow cytometry. Caspase activity was measured using specific fluorogenic substrates. Treatment with NE increased intracellular ROS level and extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 phosphorylation. Whereas the phosphorylated form of Akt was decreased. The NE-induced apoptosis was abrogated by SP600125 (a specific inhibitor of JNK). Antioxidants such as vitamin C and N-acetyl cysteine inhibited NE-induced ROS production, JNK phosphorylation, caspase activation and apoptosis. Exogenously added superoxide dismutase or catalase markedly diminished NE-induced ROS production and cell death. In conclusions, our study is the first report documenting that NE induces apoptosis in neonatal rat endothelial cells via a ROS-dependent JNK activation pathway. Antioxidants may be useful in the prevention and management of NE-mediated endothelial apoptosis during heart failure.

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Authors and Affiliations

  1. Department of Pediatrics, Taichung Veterans General Hospital, Taichung, Taiwan

    Yun-Ching Fu & Ching-Shiang Chi

  2. Department of Pediatrics, National Yang-Ming University, Taipei, Taiwan

    Yun-Ching Fu, Ching-Shiang Chi & Betau Hwang

  3. Department of Education and Research, Taichung Veterans General Hospital, 160, Section 3, Chung-Kang Road, Taichung, 40705, Taiwan, Republic of China

    Sui-Chu Yin & Shih-Lan Hsu

  4. Department of Pediatrics, Taipei Veterans General Hospital, Taipei, Taiwan

    Betau Hwang

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  1. Yun-Ching Fu
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  5. Shih-Lan Hsu
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Correspondence to Shih-Lan Hsu.

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Fu, YC., Yin, SC., Chi, CS. et al. Norepinephrine induces apoptosis in neonatal rat endothelial cells via a ROS-dependent JNK activation pathway. Apoptosis 11, 2053–2063 (2006). https://doi.org/10.1007/s10495-006-0192-8

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