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The roles of endogenous reactive oxygen species and nitric oxide in triptolide-induced apoptotic cell death in macrophages

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Abstract

Triptolide, a major active component extracted from the root of Tripterygium wilfordii Hook f, has been shown to possess potent immunosuppressive and anti-inflammatory properties. In the present report, we reported that triptolide increased the generation of reactive oxygen species (ROS) and nitric oxide (NO) and induced apoptosis of RAW 264.7 cells in a dose-dependent manner (5–25 ng/ml). The antioxidant, reduced glutathione (GSH), significantly inhibited triptolide-induced apoptosis and inhibited the degradation of Bcl-2 protein, disruption of mitochondrial membrane potential, release of cytochrome c from mitochondria into the cytosol, activation of caspase-3, and cleavage of poly-(ADP-ribose)-polymerase. The inducible nitric oxide synthase-specific inhibitor 1400w blocked triptolide-induced apoptosis, but did not alter mitochondria disruption and caspase-3 activation. These results, for the first time, implicated that the increased endogenous ROS and NO co-mediated triptolide-induced apoptosis in macrophages. ROS initiated triptolide-induced apoptosis by the mitochondria signal pathway, while the apoptotic cell death mediated by NO was not via mitochondria collapse and caspase-3 activation. In addition, combining mathematical calculation and computer simulation based on our conventional experimental results, we set and validated the apoptotic model and provided more dynamic processes of triptolide-induced apoptotic cascade in macrophages.

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Acknowledgements

We thank Dr. Zhiwei Wu (School of Dental Medicine, University of Pennsylvania) for his critical comments and grammatical correction of the manuscript. This work was supported by the “111” PROJECT and 985-PROJECT grant from Nanjing University.

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Authors and Affiliations

  1. State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, 210093, China

    Xiaofeng Bao, Jun Cui, Yuanyuan Wu, Cheng Gao, Zichun Hua & Pingping Shen

  2. Reproduction and Immunology Laboratory, Medical School of Nanjing University, Nanjing, 210093, China

    Xiaodong Han

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  1. Xiaofeng Bao
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Correspondence to Pingping Shen.

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Bao, X., Cui, J., Wu, Y. et al. The roles of endogenous reactive oxygen species and nitric oxide in triptolide-induced apoptotic cell death in macrophages. J Mol Med 85, 85–98 (2007). https://doi.org/10.1007/s00109-006-0113-x

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  • DOI: https://doi.org/10.1007/s00109-006-0113-x

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