ABSTRACT
Objective:
To investigate the improvement and possible mechanism of dl-3-n-butylphthalide (NBP) on the emotional
memory of chronic
alcoholism model
mice.
Methods:
Twenty-four
adult male C57/BL6
mice were randomly divided into
control group (CON, n=8), model group (AT, n=8),
treatment group (AT+ NBP, n=8).
Mice in AT group and AT+ NBP group were administrated with alcohol to establish chronic
alcoholism model. In the AT+ NBP group, the
mice was administrated with NBP (40 mg/kg) by gavage once a day for 14 days during the alcohol modeling period. Ang the
mice in AT group and CON group was given the same
dose of
corn oil by gavage.
Open field test was used to evaluate
anxiety-like
behavior,
tail suspension test to evaluate
depression-like
behavior,
Morris water maze and new object recognition to evaluate
memory ability, and
TUNEL staining to evaluate the number of
neuron apoptosis. The primary cultured
neurons were interfered by alcohol and NBP at the
cell level, and the
calcium concentration in the
neurons was detected by
fluorescence calcium imaging. Descriptive
analysis, t-test and one-way
ANOVA were processed by SPSS 17.0.
Results:
The results of
open field test showed that the exploration
time of AT+ NBP group was longer than that of AT group ((50.68±7.82)s, (38.50±13.93)s; t=-2.16, P<0.05)). In the
spatial memory test, the target quadrant exploration
time of AT+ NBP group was longer than that of AT group ((28.02±7.13)s, (20.98±5.58)s; t=-2.20, P<0.05). In short
memory test, the cognitive coefficient RI (0.83±0.08) of AT+ NBP group was higher than that of AT group (0.68±0.10) ( t=-3.13, P<0.05). Compared with CON group, the number of
neuron apoptosis in
prefrontal cortex in AT group increased ((17.33±2.51), (115.67±6.50); t=-24.41, P<0.001), and that in AT+ NBP group decreased compared with AT group((45.00±5.57)) ( t=14.29, P<0.001). The number of
apoptosis neurons in the
dentate gyrus of the
hippocampus in AT group (13.75±4.79) was also less than that in the AT+ NBP group (5.75±3.30) ( t=2.75, P<0.05).
Calcium concentration in
nerve cells was detected that the three concentrations of alcohol (100 mmol/L, 200 mmol/L and 300 mmol/L) led to a significant increase in the RFU within the
nerve cells (△F/F) ((1.43±0.32), (2.31±1.39), (1.21±0.73); t=-7.67, -2.85, -2.86, all P<0.05). In comparison, the changes of RFU in the three groups treated with NBP
treatment were relatively stable ((0.04±0.01), (-0.03±0.01), (-0.04±0.02); t=7.96, 2.96, 2.92, all P< 0.05).
Conclusion:
The 3-n-Butylphthalide can improve the
learning and
memory ability of chronic
alcoholism model
mice, which may be related with the inhibition of
neuron apoptosis and the influence of intracellular
calcium homeostasis.