Cardiomyocyte Apoptosis Were Inhibited by Anti-Fas Ribozyme

We investigate the effects of anti-Fas ribozyme on Fas expression and apoptosis in primary cultured Neonatal mouse ventricular myocytes. Neonatal mouse ventricular myocytes were isolated by using collagenase irrigation. A hammerhead ribozyme targeting the Fas mRNA was constructed, and transfected into Neonatal mouse ventricular myocytes via Effectene. Then Fas expression in Neonatal mouse ventricular myocytes was detected by RT-PCR and western blotting. After being treated with anti-Fas antibody (JO2), myocytes viability was measured with MTT assay. Caspase-3 proteolytic activity was detected, and cell apoptosis was measured according to Annexin V-FITC apoptosis detection kit. Fas expressed in primary mouse myocytes. Fas expression in myocytes transfected with anti-Fas ribozyme was decreased remarkably and correlated with the resistance to Fas-mediated apoptosis as determined by flow cytometry and caspase-3 proteolytic activity. Anti-Fas ribozyme can remarkably decrease the Fas expression in mouse myocytes, thus inhibit Fasmediated apoptosis in myocytes. It is suggested that anti-Fas ribozyme could significantly increase the resistance of transplanted myocytes to apoptosis and improve the survival of transplanted myocytes.