Hydrogen sulphide and mild hypothermia activate the CREB signaling pathway and prevent ischemia-reperfusion injury.

Abstract

BACKGROUND:

Both hydrogen sulphide (H2S) and mild hypothermia have been reported to prevent brain damage caused by reperfusion assault through regulating the N-methyl-D-aspartate receptor (NMDAR). However, the relationship between the two treatments and how they exert neuro-protective effects through NMDARs remain to be elucidated.

METHODS:

Transient cerebral ischemia was induced using the Pulsinelli four-vessel occlusion method. We used sodium hydrosulphide (NaHS) as the H2S donor. We randomly divided 100 Sprague-Dawley rats into five groups of 20: Sham operation group (Sh), normothermic (36-37 °C) ischemia group (NT), mild hypothermic (32-33 °C) ischemia group (mHT), normothermic ischemia combined with NaHS treatment group (NT + NaHS), and mild hypothermic ischemia combined with NaHS treatment group (mHT + NaHS). After 6 hrs of reperfusion, rats were decapitated and hippocampus samples were immediately collected. We measured NR2A (GluN1), NR2B (GluN2) and p-CREB protein levels using western blotting. We further analyzed BDNF mRNA expression by real-time PCR. Hematoxylin and eosin (HE) staining was used to examine pyramidal cell histology at the CA1 region. All statistical analyses were carried out by ANOVA and LSD t-test as implemented by the SPSS 13.0 software.

RESULTS:

In the four test groups with ischemia-reperfusion, hippocampal H2S concentration increased following treatment, and administration of NaHS further increased H2S levels. Moreover, administration of both NaHS and mild hypothermia resulted in up-regulation of NR2A and NR2B protein expressions, as well as p-CREB protein and BDNF mRNA levels. At the cellular level, NaHS and mild hypothermia groups exhibited lower damage caused by ischemia-reperfusion in the CA1 region of the hippocampus. The strongest protective effect was observed in rats treated with combined NaHS and mild hypothermia, suggesting their effects were additive.

CONCLUSION:

Our results support previous findings that hydrogen sulphide and mild hypothermia can prevent ischemia-reperfusion injury. Both treatments caused an up-regulation of NMDA receptors, as well as an elevation in p-CREB protein and BDNF mRNA levels. Thus, hydrogen sulphide and mild hypothermia may provide neuro-protective effect through activating the pro-survival CREB signaling pathway.

• Authors: Hai-bin Dai, Xiangjun Ji, Si-hai Zhu, Yi-min Hu, Li-dong Zhang, Xiao-lei Miao, Ru-Meng Ma, Man-lin Duan, Wei-yan Li
• Journal: BMC anesthesiology (BMC Anesthesiol) Vol. 15 Pg. 119 (Aug 18 2015) ISSN: 1471-2253 [Electronic] England
• PMID: 26283659 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Brain-Derived Neurotrophic Factor
• Cyclic AMP Response Element-Binding Protein
• Neuroprotective Agents
• Hydrogen Sulfide

Topics

• Analysis of Variance
• Animals
• Blotting, Western
• Brain Injuries (metabolism, prevention & control)
• Brain Ischemia (metabolism)
• Brain-Derived Neurotrophic Factor (metabolism)
• Cyclic AMP Response Element-Binding Protein (metabolism)
• Disease Models, Animal
• Hippocampus (metabolism)
• Hydrogen Sulfide (metabolism, pharmacology)
• Hypothermia, Induced
• Male
• Neuroprotective Agents (metabolism, pharmacology)
• Rats
• Rats, Sprague-Dawley
• Real-Time Polymerase Chain Reaction
• Reperfusion Injury (metabolism, prevention & control)
• Signal Transduction (drug effects, physiology)