Document Detail


HL-1 myocytes exhibit PKC and K(ATP) channel-dependent delta opioid preconditioning.
MedLine Citation:
PMID:  14559445     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Opioid preconditioning protects the myocardium against ischemia/reperfusion (IR) injury. By enhancing cardiomyocyte viability, opioids can enhance cardiac function and recovery from IR injury during acute cardiac care. The myocyte model HL-1 is an immortalized, mouse atrial cell line that expresses functional delta-opioid receptors. The HL-1 myocyte may be useful for IR injury research exploring opioid cardioprotection. MATERIALS AND METHODS: In study I, microplates of HL-1 were subjected to 10 min pre-treatment with either basal media, delta-opioid agonist DADLE(10uM), or DADLE(10uM) + delta-antagonist naltrindole (10uM). Study II treatment groups included PKC inhibitor chelerythrine (2uM), K(ATP) channel closer glybenclamide (100uM), or mitochondrial K(ATP) channel opener diazoxide (100uM) administered in various combinations followed by DADLE (10uM) or control. Microplates were subjected to normal oxygen/substrate conditions or ischemic (<1% 0(2)) and substrate deficient (10 uM 2-Deoxyglucose versus 10 mM glucose) conditions, then reperfused with normal oxygen and glucose-containing media. Microplate supernatants were subjected to lactate dehydrogenase (LDH) assay. RESULTS: Compared to untreated control, the LDH assay showed significant reduction in opioid-only pretreated groups at all time points. These effects were attenuated with delta-opioid antagonist co-administration. Co-administration of non-selective K(ATP) channel closer glybenclamide and DADLE abolished DADLE cytoprotection, while selective mitochondrial K(ATP) opener diazoxide mimicked DADLE cytoprotection Co-administration of chelerythrine and DADLE significantly reduced chelerythrine cytotoxicity. CONCLUSION: Delta-opioid preconditioning of HL-1 myocytes significantly decreased necrosis from in vitro simulated ischemia/reperfusion as measured by LDH release; this effect was reversed by delta-antagonist naltrindole. Cytoprotection was PKC and K(ATP) channel-dependent. HL-1 myocytes exhibit opioid-induced cytoprotection from IR injury, and present a novel model of pharmacologic preconditioning.
Authors:
Elisabeth M Seymour; Shu-Yung James Wu; Melissa A Kovach; Matthew A Romano; Jonathan R Traynor; William C Claycomb; Steven F Bolling
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The Journal of surgical research     Volume:  114     ISSN:  0022-4804     ISO Abbreviation:  J. Surg. Res.     Publication Date:  2003 Oct 
Date Detail:
Created Date:  2003-10-15     Completed Date:  2003-11-12     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  0376340     Medline TA:  J Surg Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  187-94     Citation Subset:  IM    
Affiliation:
Department of Cardiac Surgery, B558 MSRBII 0686, University of Michigan Medical School, Ann Arbor, MI, USA.
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MeSH Terms
Descriptor/Qualifier:
Alkaloids
Animals
Benzophenanthridines
Cell Line
Enkephalin, Leucine-2-Alanine / pharmacology
Enzyme Inhibitors / pharmacology
Heart / drug effects,  physiology*
Ischemic Preconditioning, Myocardial / methods*
Kinetics
Membrane Proteins / drug effects,  physiology*
Muscle Cells / physiology*
Myocardial Reperfusion*
Myocardium / cytology
Naltrexone / analogs & derivatives*,  pharmacology
Phenanthridines / pharmacology
Potassium Channels
Protein Kinase C / antagonists & inhibitors,  metabolism*
Receptors, Opioid, delta / drug effects,  physiology*
Grant Support
ID/Acronym/Agency:
HL58781/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Alkaloids; 0/Benzophenanthridines; 0/Enzyme Inhibitors; 0/Membrane Proteins; 0/Phenanthridines; 0/Potassium Channels; 0/Receptors, Opioid, delta; 0/mitochondrial K(ATP) channel; 111555-53-4/naltrindole; 16590-41-3/Naltrexone; 34316-15-9/chelerythrine; 63631-40-3/Enkephalin, Leucine-2-Alanine; EC 2.7.11.13/Protein Kinase C

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