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Area of Science:

  • Cell Biology
  • Cardiovascular Physiology
  • Biochemistry

Background:

  • Histones are increasingly recognized for their extracellular functions beyond nuclear roles.
  • Endothelial cell dysfunction is a critical factor in shock and cardiovascular disease.
  • Calcium signaling plays a pivotal role in cellular homeostasis and response to injury.

Purpose of the Study:

  • To investigate the mechanism of histone-induced endothelial cell damage.
  • To determine the role of store-operated calcium entry (SOCE) in histone-mediated effects.
  • To explore the influence of extracellular calcium concentrations on histone-endothelial cell interactions.

Main Methods:

  • Genetic ablation of ORAI1/2/3 channels to assess SOCE contribution.
  • Live cell video microscopy with FM1-43 membrane dye to observe membrane dynamics.
  • Exposure to varying extracellular calcium concentrations and treatment with gadolinium (Gd3+).

Main Results:

  • Histone-induced endothelial cell membrane damage was independent of SOCE.
  • Histone-induced membrane permeabilization was transient, with some cells recovering integrity.
  • Low extracellular calcium exacerbated histone-induced membrane damage, while high calcium protected cells.
  • Histone-phospholipid interactions, mediated by electrostatic forces, were identified as the primary mechanism of damage.

Conclusions:

  • Endothelial cell membrane damage by histones is primarily driven by electrostatic interactions with phospholipids, exacerbated by low extracellular calcium.
  • These findings support the clinical strategy of aggressive calcium repletion during resuscitation to stabilize cell membranes and improve outcomes in shock.
  • Understanding these mechanisms can inform therapeutic interventions for conditions involving histone-mediated endothelial injury.