Isoflurane preconditioning inhibits the effects of tissue-type plasminogen activator on brain endothelial cell in an in vitro model of ischemic stroke

Int J Med Sci. 2017 Apr 8;14(5):425-433. doi: 10.7150/ijms.18037. eCollection 2017.

Abstract

Tissue-type plasminogen activator (tPA) is the only treatment for ischemic stroke. However, tPA could induce the intracranial hemorrhage (ICH), which is the main cause of death in ischemic stroke patient after tPA treatment. At present, there is no treatment strategy to ameliorate tPA-induced brain injury after ischemia. Therefore, we investigated the effect of pre-treated isoflurane, which is a volatile anesthetic and has beneficial effects on neurological dysfunction, brain edema and infarct volume in ischemic stroke model. In this study, we used oxygen/glucose deprivation and reperfusion (OGD/R) condition to mimic an ischemic stroke in vitro. Matrix metalloproteinases (MMP) activity was measured in endothelial cell media. Also, neuronal cell culture was performed to investigate the effect of pretreated isoflurane on the neuronal cell survival after tPA-induced injury during OGD/R. Isoflurane pretreatment prevented tPA-induced MMP-2 and MMP-9 activity and suppressed tPA-triggered LRP/NF-κB/Cox-2 signaling after OGD/R. Neuronal cells, incubated with endothelial cell conditioned medium (EC-CM) after tPA + OGD/R, showed upregulation of pro-apoptotic molecules. However, neurons incubated with isoflurane-pretreated EC-CM showed increased anti-apoptotic molecules. Our findings suggest that isoflurane pretreatment could attenuate tPA-exaggerated brain ischemic injury, by reducing tPA-induced LRP/NF-κB/Cox-2 in endothelial cells, endothelial MMP-2 and MMP-9 activation, and subsequent pro-apoptotic molecule in neurons after OGD/R.

Keywords: Tissue-type plasminogen activator; endothelial cell; isoflurane; matrix metalloproteinase; neuronal cell.; oxygen/glucose deprivation.

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Brain Injuries / complications
  • Brain Injuries / drug therapy*
  • Brain Injuries / genetics
  • Brain Injuries / pathology
  • Brain Ischemia / complications
  • Brain Ischemia / drug therapy*
  • Brain Ischemia / genetics
  • Brain Ischemia / pathology
  • Culture Media, Conditioned
  • Cyclooxygenase 2 / genetics
  • Disease Models, Animal
  • Endothelial Cells / drug effects
  • Endothelial Cells / pathology
  • Glucose / metabolism
  • Humans
  • Intracranial Hemorrhages / chemically induced
  • Intracranial Hemorrhages / drug therapy
  • Intracranial Hemorrhages / genetics
  • Intracranial Hemorrhages / pathology
  • Isoflurane / administration & dosage*
  • Low Density Lipoprotein Receptor-Related Protein-1
  • Matrix Metalloproteinase 2 / genetics
  • Mice
  • NF-kappa B / genetics
  • Neurons / drug effects
  • Neurons / pathology
  • Oxygen / metabolism
  • Receptors, LDL / genetics
  • Stroke / drug therapy*
  • Stroke / genetics
  • Stroke / pathology
  • Tissue Plasminogen Activator / administration & dosage*
  • Tissue Plasminogen Activator / adverse effects
  • Tissue Plasminogen Activator / genetics
  • Tumor Suppressor Proteins / genetics

Substances

  • Culture Media, Conditioned
  • Low Density Lipoprotein Receptor-Related Protein-1
  • Lrp1 protein, mouse
  • NF-kappa B
  • Receptors, LDL
  • Tumor Suppressor Proteins
  • Isoflurane
  • Ptgs2 protein, mouse
  • Cyclooxygenase 2
  • Tissue Plasminogen Activator
  • Matrix Metalloproteinase 2
  • Glucose
  • Oxygen