Tozasertib attenuates neuronal apoptosis via DLK/JIP3/MA2K7/JNK pathway in early brain injury after SAH in rats

Cheng Yin, Guang Fu Huang, Xiao Chuan Sun, Zongduo Guo, John H. Zhang

Research output: Contribution to journalArticlepeer-review

Abstract

Background and purpose Since tozasertib is neuroprotective for injured optic nerve, this study is intended to test whether tozasertib reduces early brain injury after subarachnoid hemorrhage (SAH) in a rat model. Methods Two hundred sixteen (216) male Sprague-Dawley rats were randomly subjected to endovascular perforation model of SAH and sham group. SAH grade, neurological score, and brain water content were measured at 24 and 72 h after SAH. Dual leucine zipper kinase (DLK) and its downstream factors, JNK-interacting protein 3 (JIP3), MA2K7, p-JNK/JNK (c-Jun N-terminal kinase), and apoptosis related proteins cleaved caspase-3 (CC-3), Bim, Bcl-2, and cleaved caspase-9 (CC-9) were analyzed by western blot at 24 h after SAH. Apoptotic cells were detected by terminal deoxynucleotid transferase-deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL). DLK small interfering RNA (siRNA), JIP3 siRNA and MA2K7 siRNA, the JNK, p38MAPK, and MEK inhibitors SP600125, SB203580, and PD98059 were used for intervention. Results Tozasertib reduced neuronal apoptosis, attenuated brain edema and improved neurobehavioral deficits 24 and 72 h after SAH. At 24 h After SAH, DLK/JIP3/MA2K7/p-JNK/CC-3 expressions were elevated markedly and tozasertib reduced DLK, MA2K7/p-JNK/CC-3 expressions but enhanced JIP3 expression. In the presence of tozasertib, DLK/JIP3/MA2K7 siRNA and SP600125, SB203580 and PD98059 deteriorated the neurobehavioral deficits, brain edema and increased the expression of CC-3. SAH potentiated the expression of Bim, CC-9, and CC-3 but reduced Bcl-2, while tozasertib reduced expression of Bim, CC-9, and CC-3 but enhanced Bcl-2. Conclusions Tozasertib reduced neuronal apoptosis and improved outcome possibly via DLK/JIP3/MA2K7/JNK pathways after SAH.

Original languageEnglish
Pages (from-to)316-323
Number of pages8
JournalNeuropharmacology
Volume108
DOIs
StatePublished - Sep 1 2016

ASJC Scopus Subject Areas

  • Pharmacology
  • Cellular and Molecular Neuroscience

Keywords

  • Apoptosis
  • DLK
  • Early brain injury
  • Key words Subarachnoid hemorrhage
  • Tozasertib
  • Adaptor Proteins, Signal Transducing/antagonists & inhibitors
  • Apoptosis/drug effects
  • Injections, Intraventricular
  • MAP Kinase Kinase Kinases/antagonists & inhibitors
  • Rats
  • Male
  • Random Allocation
  • Rats, Sprague-Dawley
  • Dose-Response Relationship, Drug
  • MAP Kinase Signaling System/drug effects
  • Subarachnoid Hemorrhage/complications
  • Brain Injuries/enzymology
  • Protein Kinase Inhibitors/administration & dosage
  • Animals
  • Nerve Tissue Proteins/antagonists & inhibitors
  • Piperazines/administration & dosage

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