Activate Of ERK1/2Pathway By CaMEK Gene Via Adeno-associated Virus Serotyoe9in Cardiomyocytes

Objective:To determine whether recombinant adeno-associated virus serotyoe9with CaMEK gene transfected on myocardial cells leads to active ERK1/2and reduceI/R-induced apoptosis. Methods:Isolated and cultured neonatal rat myocardial cells,with different promoter rAAV9transfected myocardial cells isolated from the flow cyto-metry and Western blot determination of adeno-associated virus in vitro transfectionefficiency, select the appropriate promoter, and to detect rAAV9cardiac-specifictransfection. Immunofluorescence localization of detection rAAV9the mediated CaMEKgene transfection of myocardial cells by Western blot to detect the expression of ERK1/2pathway related proteins and transcription factor expression of ERK1/2pathwaysignaling molecules of the translocation activated. Star streptozotocin-induced cardiacmyocyte apoptosis by Western blot detection of each group of apoptosis related proteinexpression. Results:(1)flow cytometry results showed that the CBA promoter and theCMV promoter mediated rAAV9of transfection efficiency in cardiomyocytes(91.5±1.0)%and(73.2±1.3)%(P<0.05), by Western blot the results showed that the eGFPprotein expression rAAV9-CBA-eGFP group was significantly stronger thanrAAV9-CMV-eGFPwas group. Conclusion:Insummary, CBA start-mediated rAAV9vector specificity and efficient transfection of myocardial cells, peak expression in themyocardial cells in the first five days. rAAV9mediated CaMEKgene transfection toenhance the expression of ERK1/2pathway activity in myocardial cells, can effectivelyreduce the Star streptozotocin-induced myocardial apoptosis.