Role of protein kinase C in rat myocardial cells with anoxia-reoxygenation injury preconditioned with sufentanil / 医学研究生学报

ABSTRACT

Objective The protein kinase C ( PKC) is an essential signaling substance in the early protection of cells in pre-conditioning.This study was to investigate the role of PKC in the myocardial cells of the rat model of anoxia -reoxygenation (A-R) in-jury preconditioned with sufentanil . Methods Primary myocardial cells isolated and cultured for 5 days were allocated to a control , an A-R, a sufentanil preconditioning ( SF) , and a phorbol+sufentanil preconditioning ( PMA +SF) group.The A-R injury model was established with cultured myocardial cells from neonatal rats , which were preconditioned with sufentanil at the concentration of 0.000 3 μmol/L in the SF group or phorbol followed by sufentanil 10 minutes later in the PMA +SF group.Then the proliferation of the cells was detected , the optical density of the Cx 43 protein observed by immunofluorescence confocal technology , the apoptosis rate of the cells determined by flow cytometry, and the total Cx43 proteins calculated by Western blot. Results Cell proliferation was signifi-cantly increased in the A-R, SF, and PMA+SF groups as compared with the control (P<0.05), higher in the SF and PMA +SF than in the A-R group (0.498 0 ±0.0432 4 and 0.7240 ±0.1234 vs 0.325 8 ±0.023 5, P<0.05), and in the SF than in the PMA+SF group (P<0.05).Flow cytometry showed significantly increased rates of early , late, and total cell apoptosis in the A -R ([4.96 ±0.59], [18.77 ±0.92], and [23.73 ±0.51]%), SF ([5.86 ±0.38], [10.37 ±0.38], and [16.23 ±0.32]%), and PMA+SF group ([5.71 ±0.58], [5.54 ±0.43], [11.24 ±0.62]%) as compared with the control (P<0.05), remarkably lower in the SF and PMA +SF than in the A-R group (P<0.05), and the late and total cell apoptosis rates markedly lower in the SF than in the PMA+SF group (P<0.05). Conclusion Sufentanil has a protective effect and PKC agonists combined with sufentanil may add to the effect on myocardial cells in A -R injury.