Activated WNK3 induced by intracerebral hemorrhage deteriorates brain injury maybe via WNK3/SPAK/NKCC1 pathway

Exp Neurol. 2020 Oct:332:113386. doi: 10.1016/j.expneurol.2020.113386. Epub 2020 Jun 23.


Background: Intracerebral hemorrhage (ICH) is the common brain diseases in middle-aged and elderly people, with high disability and/or mortality rate, and is a serious public health concern. Both WNK3 kinase and the WNK3/SPAK/NKCC1 signaling pathway play an integral role in maintaining normal cell homeostasis. However, their role and underlying mechanisms in ICH-induced secondary brain injury (SBI) have yet to be elucidated.

Methods: We established an ICH model using male Sprague-Dawley (SD) rats by injecting autologous arterial blood into the unilateral basal ganglia. To establish ICH model in vitro, oxyhemoglobin (OxyHb; 20 μM) and neurons were cultured for 6 h at 37 °C, 5% CO2 atmosphere. To investigate the role of WNK3 and the WNK3/SPAK/NKCC1 signaling pathway in SBI, after genetic interventions, rotation and water maze test, brain edema and neuroinflammation were detected, and terminal-deoxynucleoitidyl transferase mediated dUTP nick end labeling (TUNEL), Fluoro-Jade C (FJC), and Nissl staining were performed.

Results: Our data showed that WNK3 expression in brain tissue were upregulated after ICH induction. In addition, silencing of WNK3 reduced neuronal apoptosis, and inflammatory responses in rats that underwent ICH. Inhibition of WNK3 expression reduced the damaged blood-brain barrier (BBB), alleviated the impaired degree of cerebral edema, and improved disruptive neurobehavioral cognition caused by ICH. Moreover, overexpression of WNK3 had the opposite effects. Finally, WNK3/SPAK/NKCC1 signaling pathway may be involved in the above-mentioned processes.

Conclusions: In conclusion, our findings showed that WNK3 and WNK3/SPAK/NKCC1 signaling pathway play a vital biological function in ICH-induced SBI. Depletion of WNK3 attenuated brain injury after ICH both in vivo and in vitro. Thus, WNK3 and WNK3/SPAK/NKCC1 signaling pathway are potential targets for treating SBI after ICH.

Keywords: Brain edema; Cell apoptosis; Intracerebral hemorrhage; Secondary brain injury; WNK3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Basal Ganglia / pathology
  • Behavior, Animal
  • Blood-Brain Barrier
  • Brain Edema / pathology
  • Cerebral Hemorrhage / genetics*
  • Cerebral Hemorrhage / pathology
  • Cerebral Hemorrhage / psychology
  • In Situ Nick-End Labeling
  • Inflammation / pathology
  • Male
  • Maze Learning
  • Protein Kinases / genetics*
  • Protein Serine-Threonine Kinases / genetics*
  • Psychomotor Performance
  • Rats
  • Rats, Sprague-Dawley
  • Rotation
  • Signal Transduction / genetics*
  • Solute Carrier Family 12, Member 2 / genetics*
  • Up-Regulation


  • Slc12a2 protein, rat
  • Solute Carrier Family 12, Member 2
  • Protein Kinases
  • Wnk3 protein, rat
  • PAS domain kinases
  • Protein Serine-Threonine Kinases