Neuroprotective effect of G14-humanin on global cerebral ischemia/reperfusion by activation of SOCS3 - STAT3 - MCL-1 signal transduction pathway in rats

Neurol Res. 2017 Oct;39(10):895-903. doi: 10.1080/01616412.2017.1352187. Epub 2017 Jul 18.

Abstract

Objective: Humanin (HN) has been identified to suppress neuron death. Gly14-HN (HNG), as a variant of HN, can decrease infarct volume after ischemia/reperfusion (I/R) injury. This study aimed to investigate the neuroprotective mechanism of HNG on global cerebral I/R (GI) in rats.

Methods: Rats were randomly divided into 13 groups: Sham group, GI groups and HNG groups. Both GI group and HNG groups included six time points (1, 3, 6, 12, 24, and 72 h). At 24 h after reperfusion, Nissl staining was used to observe positive neurons, and p-STAT3, MCL-1, SOCS3, Bax and Caspase-3 in different groups were detected by immunohistochemistry. qRT-PCR and western blot were used to evaluate the expression of STAT3, p-STAT3, MCL-1, and SOCS3.

Results: The immunohistochemistry also showed a significant increase in Bax (0.29 ± 0.007 vs. 0.22 ± 0.007, P < 0.01) and Caspase-3 (0.24 ± 0.02 vs. 0.18 ± 0.006, P < 0.01) in GI group compared with Sham group, while Bax (0.26 ± 0.01 vs. 0.29 ± 0.008, P < 0.01) and Caspase-3 (0.20 ± 0.008 vs. 0.24 ± 0.02, P < 0.01) were significantly decreased by HNG-treatment compared with GI group. Along with immunohistochemistry, western blot and qRT-PCR indicated that the protein and mRNA levels of STAT3, MCL-1, and SOCS3 were up-regulated after administration of HNG at six time points after global cerebral I/R in rat.

Conclusion: HNG might exert neuroprotective effects through alleviating apoptosis and activating of SOCS3 - STAT3 - MCL-1 signal transduction pathway. Highlights (1) Cerebral ischemia led to neuronal loss in hippocampal CA1 region of rats. (2) HNG had neuroprotective effects on ischemia/reperfusion rats. (3) The protective effect of HNG might be related to the SOCS3 - STAT3 - MCL-1 pathway.

Keywords: G14-humanin; STAT3; global cerebral ischemia/reperfusion; neuroprotective.

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Apoptosis / physiology
  • Brain Ischemia / drug therapy*
  • Brain Ischemia / metabolism
  • Brain Ischemia / pathology
  • Caspase 3 / metabolism
  • Disease Models, Animal
  • Male
  • Myeloid Cell Leukemia Sequence 1 Protein / metabolism
  • Neuroprotective Agents / pharmacology*
  • Nissl Bodies / drug effects
  • Nissl Bodies / metabolism
  • Nissl Bodies / pathology
  • Peptides / pharmacology*
  • RNA, Messenger / metabolism
  • Random Allocation
  • Rats, Sprague-Dawley
  • Reperfusion Injury / drug therapy*
  • Reperfusion Injury / metabolism
  • Reperfusion Injury / pathology
  • STAT3 Transcription Factor / metabolism
  • Signal Transduction / drug effects
  • Suppressor of Cytokine Signaling 3 Protein / metabolism
  • bcl-2-Associated X Protein / metabolism

Substances

  • Bax protein, rat
  • Gly(14)-Humanin
  • Mcl1 protein, rat
  • Myeloid Cell Leukemia Sequence 1 Protein
  • Neuroprotective Agents
  • Peptides
  • RNA, Messenger
  • STAT3 Transcription Factor
  • Socs3 protein, rat
  • Stat3 protein, rat
  • Suppressor of Cytokine Signaling 3 Protein
  • bcl-2-Associated X Protein
  • Casp3 protein, rat
  • Caspase 3