Influence of the nitric oxide-donor of sodium nitroprusside on the expression of hippocampal neurons gene cpp32

Yongjun LIU; Haifeng ZHANG; Qifeng ZHU; Ailing LIANG

ABSTRACT

BACKGROUND:

Alzheimer disease is a senile degenerative disease of nervous system. Neuron apoptosis is regarded as one of possible reasons,and neuron culture in vitro is a common method to research the mechanism of apoptosis.

OBJECTIVE:

To observe the influences of nitric oxide-donor, sodium nitroprusside (SNP), on the expression of gene cpp32 in the cultured hippocarnpal neurons in vitro.

DESIGN:

A randomized controlled animal experiment.

SETTING:

Institute of Biochemistry and Molecular Biology in Guangdong Medical College.MATERIALS The experiment was carried out in the Institute of Biochemistry and Molecular Biology, Guangdong Medical College, and newborn (＜ 24 hours) Sprague-Dawley rats were used.

METHODS:

The hippocampl neurons of rats were primarily cultured, and then treated with SNP of different terminal concentrations (0, 25, 50, 100,200, 400 μmol/L) for 24 hours, and the expressions of mRNA and protein were analyzed with RT-PCR and Western blot respectively. The hippocampl neurons of rats were treated with SNP of different terminal concentrations (0, 25, 50, 100, 200, 400 μmol/L) for 12 hours, and the activity of CPP32 enzyme was detected with CPP32 activity detected kit.MAIN OUTCOME

MEASURES:

The expressions cpp32 mRNA and CPP32 protein and activity of CPP32 were detected.

RESULTS:

The cpp32 mRNA expression was unchanged as the increasing dose of SNP, but the pro-CPP32 was activated and the activity of CPP32 was increased significantly at 50 μmol/L SNP which was 3.02 times of that in the control group, and reached to the maximal value at 100 μmol/L which was 3.47 times of that in the control group.

CONCLUSION:

SNP cannot increase the cpp32 mRNA expression, but can increase degradation of pro-CPP32 and activate CPP32.