Preliminary study on functions of AKAP1 in mouse primary cardiomyocytes

AIM To investigate the functions of A kinase anchoring protein 1 (AKAP1) in cardiomyocytes by construction of AKAP1 recombinant adenovirus vector using AdEasy adenovirus vector system. METHODS AKAP1 was PCR-amplified using murine cDNA as a template and then recombinant Ad plasmid pAdEasy-AKAP1 was constructed. The recombinant adenovirus Ad-AKAP1 was packaged and amplified by using 293A cells. The virus titer was determined by green fluorescent protein labeling method. AKAP1 and Caspase-3 expression levels in mouse primary cardiomyocytes and diabetic cardiomyopathy model cells infecting recombinant adenovirus were detected by Western blot. The levels of ROS in mouse primary cardiomyocytes and the effect of AKAP1 on apoptosis of diabetic cardiomyopathy cells exposed to high glucose were detected by flow cytometry. RESULTS The recombinant adenovirus carrying AKAP1 gene was constructed and high titer recombinant adenovirus of 5×10¹⁰ pfu/ml was obtained. The recombinant adenovirus can infect primary myocardial cells and diabetic cardiomyopathy model H9c2 cells and mediates AKAP1 overexpression (P<0.05). Compared with the negative control group, overexpression of AKAP1 could inhibit the formation of ROS in cardiomyocytes (P<0.05) and could inhibit the apoptosis of cardiac myocytes and the activation of Caspase-3 (P<0.01) under high glucose and fat condition. CONCLUSION Overexpression of AKAP1 has a potential protective effect on cardiomyocytes under high glucose and fat condition, which plays an important role in myocardial function. The mechanism may be related to which AKAP1 inhibited the formation of ROS, inhibited the activation of Caspase-3 and inhibition the apoptosis in cardiomyocytes.