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J Korean Soc Emerg Med > Volume 23(1); 2012 > Article
Journal of The Korean Society of Emergency Medicine 2012;23(1): 111-119.
The Neuroprotective Effect of delta-opioid Receptor Stimulation with [D-Ala2, D-Leu5] Enkephalin Against Ischemic Neuronal Injury
Hoon Kim, Suk Woo Lee, Jung Soo Park, Jin Hong Min, Mun Ki Min
1Department of Emergency Medicine, College of Medicine, Chungbuk National University, Korea. nichekh2000@chungbuk.ac.kr
2Department of Emergency Medicine, Busan National University Yangsan Hospital, Korea.
Oxygen is indispensable for survival and aerobic metabolism in all mammalian cells. Inadequate oxygen triggers a multifaceted cellular response negatively impacting important physiological functions which are observed in clinical diseases such as stroke, drowning, cardiac arrest, hazardous gas poisoning, myocardial infarction and vascular dementia. In this study, we investigated the neuroprotective effect of a synthetic delta-opioid agonist, [D-Ala2, D-Leu5] enkephalin (DADLE), and its role in ischemic neuronal injury.
This experiment was conducted in vitro using a primary culture of rat cortical neurons. Ischemia induction was performed using a hypoxic chamber. To test the degree of neuronal viability, as protected by delta-opioid stimulation with DADLE under ischemia, we used three independent approaches including a lactate dehydrogenase assay, MTT assay, and an immunofluorescent staining assay for viable cells. In addition, the gene expressions of caspase-3 and heat shock protein 70 were analyzed using real-time PCR.
Incubation of the cortical neurons with DADLE protected them from ischemia-induced cytotoxicity, as observed by all three independent viability assays. Also, we found that its neuroprotective effect might be related with suppression of the caspase-3 gene.
The results of this study suggested that DADLE exhibits a neuroprotective effect against ischemia-induced neuronal cell death.
Key words: Ischemia, [D-Ala2, D-Leu5] enkephalin (DADLE), Cortical neurons
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