Inhibition of mouse hepatocyte apoptosis via anti-Fas ribozyme

AIM: TO investigate the effects of anti-Fas ribozyme on Fas expression and apoptosis in primary cultured mouse hepatocytes.METHODS: Mouse hepatocytes were isolated by using collagenase irrigation. A hammerhead ribozyme targeting the Fas mRNA was constructed, and transfected into mouse hepatocytes via Effectene. Then Fas expression in mouse hepatocytes was detected by RT-PCR and western blotting.After being treated with anti-Fas antibody (JO2), hepatocytes viability was measured with MTI- assay. Caspase-3 proteolytic activity was detected, and cell apoptosis was measured according to Annexin V-FITC apoptosis detection kit.RESULTS: Fas expressed in primary mouse hepatocytes.Fas expression in hepatocytes 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.CONCLUSION: AnU-Fas ribozyme can remarkably decrease the Fas expression in mouse hepatocytes, thus inhibit Fasmediated apoptosis in hepatocytes, It is suggested that anti-Fas ribozyme could significantly increase the resistance of transplanted hepatocytes to apoptosis and improve the survival of transplanted hepatocytes,

AIM:To investigate the effects of anti-Fas ribozyme on Fas expression and apoptosis in primary cultured mouse hepatocytes. METHODS:Mouse hepatocytes were isolated by using collagenase irrigation.A hammerhead ribozyme targeting the Fas mRNA was constructed,and transfected into mouse hepatocytes via Effectene.Then Fas expression in mouse hepatocytes was detected by RT-PCR and western blotting. After being treated with anti-Fas antibody(JO_2),hepatocytes 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. RESULTS:Fas expressed in primary mouse hepatocytes. Fas expression in hepatocytes 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. CONCLUSION:Anti-Fas ribozyme can remarkably decrease the Fas expression in mouse hepatocytes,thus inhibit Fas- mediated apoptosis in hepatocytes.It is suggested that anti-Fas ribozyme could significantly increase the resistance of transplanted hepatocytes to apoptosis and improve the survival of transplanted hepatocytes.