The ROCK inhibitor Y-27632 ameliorates blood-spinal cord barrier disruption by reducing tight junction protein degradation via the MYPT1-MLC2 pathway after spinal cord injury in rats

Brain Res. 2021 Dec 15:1773:147684. doi: 10.1016/j.brainres.2021.147684. Epub 2021 Oct 9.

Abstract

The blood-spinal cord barrier (BSCB) is a physiological barrier between the blood and spinal cord parenchyma. This study aims to determine whether Y-27632, a Rho-associated protein kinase (ROCK) inhibitor, can protect the BSCB using in vivo models. The Evans blue fluorescence assay was used to detect leakage of the BSCB. Western blotting was used to define alterations in ROCK-related and tight junction (TJ) protein expression. Immunofluorescence triple-staining was used to evaluate histologic alterations in TJs. Locomotor function was evaluated using the open-field test, the Basso-Beattie-Bresnahan score, and footprint analysis. Two peaks of BSCB leakage after spinal cord injury (SCI) occurred at 24 h and 5 days. The ROCK inhibitor reduced the BSCB leakage at the second peak after SCI. Moreover, the ROCK inhibitor ameliorated the integrity of the BSCB and improved motor function recovery after SCI by regulating the phosphorylation of myosin phosphatase subunit-1 (MYPT1) and cofilin. ROCK inhibitors might protect the BSCB, which provides a new strategy for transitioning SCI treatment from the bench to bedside.

Keywords: Blood-spinal cord barrier; Evans blue; ROCK inhibitor; Spinal cord injury; Tight junction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amides / pharmacology*
  • Animals
  • Blood-Brain Barrier / drug effects*
  • Blood-Brain Barrier / metabolism
  • Blood-Brain Barrier / pathology
  • Cardiac Myosins / metabolism
  • Enzyme Inhibitors / pharmacology*
  • Male
  • Myosin Light Chains / metabolism
  • Phosphorylation / drug effects
  • Protein Phosphatase 1 / metabolism
  • Proteolysis / drug effects
  • Pyridines / pharmacology*
  • Rats
  • Rats, Sprague-Dawley
  • Signal Transduction / drug effects*
  • Spinal Cord Injuries / metabolism*
  • Spinal Cord Injuries / pathology
  • Tight Junction Proteins / metabolism*
  • Tight Junctions / metabolism*
  • rho-Associated Kinases / antagonists & inhibitors

Substances

  • Amides
  • Enzyme Inhibitors
  • Myosin Light Chains
  • Pyridines
  • Tight Junction Proteins
  • myosin light chain 2
  • Y 27632
  • rho-Associated Kinases
  • Ppp1r12a protein, rat
  • Protein Phosphatase 1
  • Cardiac Myosins