The Role of Neutrophil Extracellular Traps in Early Microthrombosis and Brain Injury After Subarachnoid Hemorrhage in Mice

Abstract

Microthrombosis plays an important role in secondary brain injury after experimental subarachnoid hemorrhage (SAH), but the specific mechanism of microthrombosis remains unclear. The purpose of this study was to investigate the role of neutrophil extracellular traps (NETs) in microthrombosis after SAH. SAH was induced in male C57BL/6 mice using an endovascular perforation technique. The marker protein of NETs, citrullinated histone H3 (CitH3), was significantly elevated in the cerebral cortex after SAH, and was co-labeled with microthrombi. Both depletion of neutrophils by anti-Ly6G antibody and DNase I treatment significantly reduced the formation of NETs and microthrombi, and ameliorated neurological deficits, brain edema, BBB disruption, and neuronal injury at 24 h after SAH induction. Cerebral hypoperfusion in the first hours after SAH is a major determinant of poor neurological outcome; in this study, we found that DNase I treatment significantly improved the restoration of early cortical perfusion after SAH. In addition, DNase I treatment also significantly attenuated cerebrospinal fluid (CSF) flow after SAH, which was associated with the diffusion barrier caused by microthrombi in the paravascular space after SAH. In conclusion, NETs are associated with early microthrombosis after SAH; they may be a novel therapeutic target for early brain injury (EBI) after SAH.

Keywords: Early brain injury; Microthrombosis; Neutrophil extracellular traps; Subarachnoid hemorrhage.