Objective To investigate the uppressive effects of par-4 antisense oligodeoxyn ucleotide on the up-
regulation of intracellular
calcium concentration in PC12 c ell induced by glutamate and its anti-apoptosis effects. Methods Cationic lipid-mediated par-4 antisense oligodeoxynucleotide (par-4-AS-ODN) was transfected into
PC12 cells and followed by glutamate for treatment. Misma tch oligodeoxy-nucleotide (MS-ODN) was used as the control. Morphological ass essment and evaluation of the anti-apoptosis effects of par-4-AS-ODN on PC12 cells were performed by laser scanning confocal microscopy by double staining o f the cells with Hoechest 33258/propidium iodide (Hoe/PI) and flow cytometry res pectively. The mRNA and protein levels of calpain 10 and par-4 were measured b y RT-PCR and Western blot. Intracellular calcium concentration was determined by using laser scanning confocal microscope with Fura-2/AM as the fluorescent d ye. Results Par-4-AS-ODN repress the increase of par-4 protein in PC12 cell( 52.3±5.0 vs 90.0±3.2, P<0.01).Par-4-AS-ODN significantly inhibit ed the apoptosis of PC12 cells induced by glutamate(53% vs 31%, P<0.01). Par-4-AS-ODN significantly suppress the up-
regulation of intracellular calc ium concentration in PC12 cells induced by glutamate(Rate of fluorescent densit y: 167.9±32.4 vs 228.8±36.8, P<0.01).Par-4-AS-ODN inhibited the increase of calpain 10 mRNA in PC12 cells induced by glutamate(46.3±3.7 vs 3 4.8±2.1, P<0.01). Conclusions par-4-AS-ODN enables to inhibit apoptosis of PC12 cells induced by glutamate. The mechanism of the inhibition may be closely related to suppressi on of the up-regulation of intracellular calcium concentration and calpain tran scription expression.