Role of opioid receptors and PI3K/Akt and ERK signaling pathways in reduction of hypoxia/reoxygenationinduced injury to cardiomyocytes by remifentanil preconditioning in rats / 中华麻醉学杂志

ABSTRACT

Objective To evaluate the role of opioid receptors and phosphatidylinositol 3-kinase/proteinserine-threonine kinases (PI3K/Akt) and extracellular signal-regulated kinase (ERK) signaling pathways in reduction of hypoxia/reoxygenation (H/R)-induced injury to cardiomyocytes by remifentanil preconditioning in rats.Methods Primary cardiomyocytes were obtained from adult male Sprague-Dawley rats and cultured in DMEM culture medium.The cells were seeded in 48-well plates (density 2 × 104 cells/ml,500 μl/well) and randomly divided into 12 groups (n =9 each)control group (group C),group H/R,hypoxia preconditioning group (group HPC),remifentanil preconditioning (RPC) group,naltrindole (δ receptor antagonist) + RPC group,nor-binaltorphimine (κ receptor antagonist) + RPC group (BNI + RPC group),wortmannin (PI3K inhibitor) + RPC group (W+ RPC group),PD98059 (ERK inhibitor) + RPC group (PD + RPC group),NTD group,BNI group,W group and PD group.In group H/R,the cardiomyocytes were exposed to 90 min of hypoxia,followed by 120 min of reoxygenation.In group HPC,the cardiomyocytes were exposed to 10 min of hypoxia,followed by 30 min of reoxygenation before H/R.In group RPC,the cardiomyocytes were preconditioned with remifentanil with the final concentration of 1 μmol/L for 10 min,followed by 30 min routine culture before H/R.In NTD + RPC,BNI + RPC,W + RPC and PD + RPC groups,naltrindole 5μmol/L (final concentration),nor-binaltorphimine 5 μmol/L (final concentration),wortmannin 0.1 μmol/L (final concentration) and PD98059 30μmol/L (final concentration)were added,respectively,and then the cells were coincubated with remifentanil for 10 min,followed by 30 min routine culture before H/R.The viability of cardiomyocytes,cell apoptosis and activity of lactate dehydrogenase (LDH) in the culture medium were detected.The apoptosis rate (AR) was calculated.Results Compared with group C,the viability of cardiomyocytes,AR and activity of LDH in the culture medium were significantly increased in group H/R (P ＜ 0.05).Compared with group H/R,the viability of cardiomyocytes,AR and activity of LDH in the culture medium were significantly decreased in HPC,RPC and BNI + RPC groups (P ＜ 0.05),and no significant changes were found in the parameters mentioned above in NTD + RPC,W + RPC,PD + RPC,NTD,BNI,W,and PD groups (P ＞ 0.05).The viability of cardiomyocytes was significantly lower,and the AR and activity of LDH in the culture medium were higher in NTD + RPC,BNI + RPC,W + RPC,and PD + RPC groups than in RPC group (P ＜ 0.05).Conclusion Remifentanil preconditioning activates PI3K/Akt and ERK signaling pathways possibly through activating δ opioid receptors thus attenuating H/R-induced injury to cardiomyocytes in rats.