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Related Experiment Video

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Detection of Inflammasome Activation and Pyroptotic Cell Death in Murine Bone Marrow-derived Macrophages
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Nuclear DAMP complex-mediated RAGE-dependent macrophage cell death.

Ruochan Chen1, Sha Fu2, Xue-Gong Fan2

  • 1Department of Surgery, University of Pittsburgh, Pittsburgh, PA 15213, USA; Department of Infectious Diseases and State Key Lab of Viral Hepatitis, Xiangya Hospital, Central South University, Changsha, Hunan 410008, China.

Biochemical and Biophysical Research Communications
|February 17, 2015
PubMed
Summary
This summary is machine-generated.

Nuclear DNA-HMGB1-histone complexes (nDCs) induce macrophage death by promoting TNFα release via RAGE signaling. This highlights a novel mechanism linking DAMPs and inflammation in immune cell demise.

Keywords:
AktDNAHMGB1HistoneRAGETNFα

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Highly Efficient Transfection of Human THP-1 Macrophages by Nucleofection
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Highly Efficient Transfection of Human THP-1 Macrophages by Nucleofection

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Area of Science:

  • Immunology
  • Cell Biology
  • Molecular Biology

Background:

  • High mobility group box 1 (HMGB1), histones, and DNA are nuclear components with extracellular damage-associated molecular pattern (DAMP) functions.
  • Extracellular nuclear DNA-HMGB1-histone complexes (nDCs) act synergistically as DAMP complexes.
  • The impact of nDCs on immune cells is not well understood.

Purpose of the Study:

  • To investigate the effects of nDCs on immune cells and cancer cells.
  • To elucidate the signaling pathways involved in nDC-induced cell death.

Main Methods:

  • Treatment of macrophages (RAW264.7, peritoneal) and cancer cells (HCT116, HepG2, Hepa1-6) with nDCs.
  • Assessment of cell survival, TNFα production, and reactive oxygen species (ROS).
  • Evaluation of the roles of RAGE, TLR-4, TLR-2, and Akt signaling.
  • Intervention with RNAi for RAGE, N-Acetyl-l-cysteine (antioxidant), and TNFα neutralizing antibody.

Main Results:

  • nDCs reduced the survival of macrophages but not cancer cells.
  • nDCs induced TNFα release, leading to ROS-dependent apoptosis and necrosis.
  • Receptor for advanced glycation end products (RAGE) signaling was essential for TNFα release and cell death, while TLR-4 and TLR-2 were not involved.
  • RAGE depletion, antioxidant treatment, and TNFα neutralization attenuated nDC-induced cell death.

Conclusions:

  • nDCs trigger macrophage cell death through RAGE-dependent TNFα production and ROS generation.
  • This study reveals novel signaling pathways connecting nDCs, inflammation, and macrophage apoptosis.
  • Findings suggest a potential role for nDCs in inflammatory processes affecting macrophages.