Role Of PKC Signaling Pathways And Cx43 With Anoxia/Reoxygenation By Sufentanil Preconditioning In Rat Cardiomyocytes

Objective To evalute the Role of PKC signaling pathways and Cx43 with anoxia/reoxygenation by sufentanil preconditioning in rat cardiomyocytes.Methods Primary culltured cardiomyocytes, Using the 5d cardiomyocytes divided into eight groups: Normal control group(C group): culture process does not give any treatment. Hypoxia/reoxygenation injury group(I/R group): hypoxia 2h, reoxygenation 1h.Morphine preconditioning group(MF group): morphine incubate 15 min and remove the drug culture 20 min, the other do the same with the I/R group. Sufentanil preconditioning group(SF group): sufentanil incubate 15 min, remove the drug culture 20 min, the other do the same with the I/R group.PKC agonists PMA plus morphine preconditioning group(PMA+MF group): PMA incubated 10 min,then use morphine pretreat, the other do the same with the I/R group.PKC inhibitor Rottlerin plus morphine preconditioning group(ROT+MF group): Rottlerin incubated 10 min, then use morphine pretreat, the other do the same with the I/R group. PKC agonists PMA plus sufentanyl preconditioning group(PMA+SF group): PMA incubated 10 min, then use sufentanil pretreat,the other do the same with the I/R group. PKC inhibitor Rottlerin plus sufentanil preconditioning group(ROT+SF group): use Rottlerin incubated 10 min, then use sufentanil to pretreat, the other do the same with the I/R group. and each group made those treatment to detect cell survival. Confocal immunofluorescence detect connexin Connexin 43(Cx43) the average optical density(AOD),Western-blot detect expression of Cx43 total protein and its phosphorylation.Results Compared with C group, other all groups of the cardiomyocytes viability, Cx43 AOD,Cx43 total protein and P-Cx43 were decreased; Compared with I/R group, ROT plus MF group, ROT plus SF, MF group, SF group,PMA plus MF group, PMA plus SF group cardiomyocytes viability, Cx43 AOD,Cx43 total protein and P-Cx43 were increased(P<0.05);Compared with the MF group,ROT plus MF group cardiomyocytes viability,Cx43 AOD and Cx43 total protein were decreased(P<0.05), SF group cardiomyocytes viability, Cx43 AOD and Cx43 total protein were decreased(P<0.05),SF group P-Cx43 was increased(P<0.05), PMA plus MF group cardiomyocytes viability, Cx43 AOD, Cx43 total protein and P-Cx43 were increased(P<0.05); Compared with the SF group, ROT plus SF cardiomyocytes viability, Cx43 AOD,Cx43 total protein and P-Cx43 were decreased(P<0.05),PMA plus SF group cardiomyocytes viability,Cx43 AOD, Cx43 total protein and P-Cx43 were increased(P<0.05).Conclusion Pretreatment of morphine and sufentanil preconditioning can reduce injury of myocardial hypoxia/reoxygenation.and the cardiomyocytes' protective effect of Sufentanil are weaker than morphine, which may be related to cardiomyocytes activated PKC signaling pathways,which through PKC signaling pathway regulates the expression of Cx43 and its phosphorylation.