Protective Effects of the Delta Opioid Peptide [ D ‐ A la2, D ‐ L eu5]enkephalin in an iEx Vivo/i Model of Ischemia/Reperfusion in Brain Slices

摘要

Summary Introduction The delta opioid peptide [ D ‐ A la2, D ‐ L eu5]enkephalin ( DADLE ) plays a key role in neuronal protection against both hypoxic and ischemic conditions. However, the cellular mechanisms of action of DADLE under these conditions remain unclear. Methods Ischemia was simulated with perfusing the brain slices with glucose‐free artificial cerebrospinal fluid. Apoptosis was examined using an in situ cell death detection kit and expressed as the percentage of positively labeled neurons relative to total number of neurons. PCR was performed by adding c DNA , 5 pm dNTP , 1 μL Taqase, and primers. PCR products were separated with electrophoresis, stained with ethidium bromide, and visualized under ultraviolet light. Aims To investigate the potential effects of DADLE in an ex vivo model of cerebral ischemia/reperfusion. Results DADLE attenuated lactic dehydrogenase release and neuronal apoptosis in a concentration‐dependent manner. The protective effects of DADLE were attenuated by representative selective delta2, but not delta1 opioid antagonists. Treatment with PD 98059, a selective inhibitor of ERK kinase ( MEK ), also blocked the protective effect of DADLE as well as ERK phosphorylation induced by DADLE . Conclusions Endogenous opioid peptides could promote cell survival via delta2 opioid receptors, possibly through the downstream MEK ‐ ERK pathway.