Anti-apoptotic effect of morphine-induced delayed preconditioning on pulmonary artery endothelial cells with anoxia/reoxygenation injury.
- Author:
Wen-gang DING
1
;
Hua-cheng ZHOU
;
Xiao-guang CUI
;
Wen-zhi LI
;
Yue-ping GUO
;
Bing ZHANG
;
Wei LIU
Author Information
- Publication Type:Journal Article
- MeSH: Analgesics, Opioid; pharmacology; Animals; Apoptosis; drug effects; Cell Hypoxia; Cells, Cultured; Endothelial Cells; cytology; drug effects; metabolism; Enzyme Inhibitors; pharmacology; Glyburide; pharmacology; In Situ Nick-End Labeling; Morphine; pharmacology; NG-Nitroarginine Methyl Ester; pharmacology; Naloxone; pharmacology; Narcotic Antagonists; pharmacology; Nitric Oxide; metabolism; Nitric Oxide Synthase Type III; antagonists & inhibitors; genetics; metabolism; Oxygen; pharmacology; Pulmonary Artery; cytology; RNA, Messenger; genetics; metabolism; Reverse Transcriptase Polymerase Chain Reaction; Swine
- From: Chinese Medical Journal 2008;121(14):1313-1318
- CountryChina
- Language:English
-
Abstract:
BACKGROUNDOpioid preconditioning (PC) reduces anoxia/reoxygenation (A/R) injury to various cells. However, it remains unclear whether opioid-induced delayed PC would show anti-apoptotic effects on pulmonary artery endothelial cells (PAECs) suffering from A/R injury. The present study was conducted to elucidate this issue and to investigate the potential mechanism of opioid-induced delayed PC.
METHODSCultured porcine PAECs underwent 16-hour anoxia followed by 1-hour reoxygenation 24 hours after pretreatment with saline (NaCl; 0.9%) or morphine (1 micromol/L). To determine the underlying mechanism, a non-selective K(ATP) channel inhibitor glibenclamide (Glib; 10 micromol/L), a nitric oxide (NO) synthase blocker NG-nitro-L-arginine methyl ester (L-NAME; 100 micromol/L), and an opioid receptor antagonist naloxone (Nal; 10 micromol/L) were given 30 minutes before the A/R load. The percentage of apoptotic cells was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. eNOS mRNA level was measured by real-time polymerase chain reaction (PCR). NO content of PAECs supernatants was measured with the Griess reagent.
RESULTSCompared to the A/R PAECs, morphine-induced delayed PC significantly reduced PAECs apoptosis ((18.1 +/- 1.9)% vs (5.5 +/- 0.3)%; P < 0.05), increased NO release ((11.4 +/- 1.3) micromol/L vs (20.5 +/- 2.1) micromol/L, P < 0.05), and up-regulated eNOS gene expression nearly 9 times (P < 0.05). The anti-apoptosis effect of morphine was abolished by pretreatment with Glib, L-NAME and Nal, but the three agent-selves did not aggravate the A/R injury. Furthermore, L-NAME and Nal offset the enhanced release of NO caused by pretreatment with morphine.
CONCLUSIONSMorphine-induced delayed PC prevents A/R injury of PAECs. This effect may be mediated by activation of K(ATP) channel via opioid receptor and NO signaling pathways.