Download THE EFFECTS OF PIRACETAM ON COGNITIVE PERFORMANCE

THE EFFECTS OF PIRACETAM ON COGNITIVE PERFORMANCE IN ADULT
FEMALE MICE (Mus musculus).
Behrad Salehian, Karla Hernandez, and ln Kyung Kim.
Department of Biological Science
Saddleback College
Mission Viejo, CA 92692.
Piracetam is a nootropic drugs that has been widely studied for its ability to enhance
memory and learning factors in the cerebrum. It is often referred to as a “smart drug”
because it boosts verbal memory and allows for deeper cognitive retention. Piracetam is a
non-toxic and non addictive supplement with few known side effects. In this experiment,
three groups of 6 female mice (Mus musculus) were being tested on their learning ability to
complete a constructed branching maze and evaluated on the basis of completion times. All
three groups were trained and also timed to run the maze three times for the first week
without any given drug. The control group was orally fed with 0.5 mL of water. The two
experimental groups consisted of low (0.5mg) and high (1.5 mg) dosages were given a 0.5 ml
water solution of Piracetam. This feeding pattern was continued daily for the next two
weeks as they were being tested in the maze every other day. The average time for nondosage group was 77.2±5.31 seconds (±SEM, N=6) while it was 56.3±6.16 seconds for low
dosage group (±SEM, N=6) and 46.0±6.98 seconds for high dosage group (±SEM, N=6).
The study conducted yielded that there was a significant difference between all three
groups (P=0.002, ANOVA). Further analysis using Bonferroni Post-Hoc revealed that there
was only a significant difference between high dosage and the control group (p<0.05).
Introduction
Piracetam is a commonly used and known nootropic drug. The term nootropic meaning to
“mind-bend” was coined in 1972 by Professor Corneliu E. Giurgea. He characterized such drugs
for their ability to activate brain mechanisms that triggered memory improvements, enhance
daily brain functions, and stimulating nerve activity (Giurgea, 1982). Of these brain mechanisms,
Piracetam allows for an increase of calcium influx in neural cells and an increase of L-glutamate
neurotransmitter (Bartsch et al, 1997). L-Glutamate is a common excitatory neurotransmitter
found in the central nervous system. Nerve impulses activate the release of glutamate from a presynaptic cell which binds to a NMDA glutamate receptor located on the post-synaptic cell;
stimulating it. Glutamate is responsible for cognitive functions in learning, memorization, and
motor functions (Malykh and Sadaie, 2010). In a research study performed in 1999 researchers
showed that Piracetam enhanced the facilitation of glutamate on to NMDA glutamate receptors
by binding to ligand’s on the glutamate. They concluded that this could be a therapeutic
approach to treat patients with Schizophrenia (Amini-Nooshabadi H et al, 1999). When
combined these two mechanism’s have been shown to increase the overall activity of the
hemispheres of the cerebrum in working collaboration (Abdel-Salam, et al, 2011).
The main focus of this investigation is to determine the effects of Piracetam® on the
cognitive performance of adult female mice (Mus musculus). In a previous study done by
Bartsch et. al (1997) showed that Piracetam increased membrane fluidity in the brain of aged
rats that were given Piracetam orally for several weeks. They concluded that increased
membrane fluidity is especially important for cell regeneration, receptor function, and brain
performance. It is hypothesized that female mice (Mus musculus) given higher dosages of
Piracetam will show relatively more motor functions and memory capabilities than the control
group and the low dosage group.
Methods and Materials
Subjects
In this experiment eighteen six to seven week old female feeding mice were bought from
PetCo in Mission Viejo, California. The mice were grouped into three separate groups containing
six mice. Each group was housed in individual ten gallon glass tanks in Mission Viejo,
California. All three housing tanks were identical in dimensions. Each housing tank measured
50.8 x 25.4 x 30.5 centimeters with a wire cover top. Each of the tanks had Aspen wood and
paper shavings for bedding, a suspended water bottle, a bowl for food, a small paper house made
from shoe boxes, and a small mesh exercise wheel. All mice were acclimated into their housing
tanks where they were fed Kaytee Forti-diet pro health nuggets and fresh water daily.
Piracetam was purchased from a health online store that offers nutritional supplements
(www.healthsuperstore.com).
Procedures
On the first day, the mice were grouped into three groups (No dosage/control group, Low
dosage, and High dosage). They would remain in these groups until the end of the experiment.
Before the experiment all mice were pre-weighed. Piracetam dosages were determined by weight
for each individual mouse. Formula used for calculations:
dosage (mg)/kg x weight (kg) = mg of dosage per mouse.
The suggested minimal dosage (25mg per kg body weight) and the maximal dosage
(75mg per kg bodyweight) were respectively used for low and high dosage groups. After final
calculations, low dosage experimental group received 5 mg of Piracetam per mouse and 15 mg
of Piracetam per mouse for high dosage group. During week one each mouse were taken from
their assigned groups and placed into a branched maze and timed for completion to the end of the
maze. The maze was constructed from plastic Lego building blocks that measured 86.4 by 61
centimeters in diameter and placed on a white board. This would allow for easy clean up to
eliminate the odor produce by the mice. No drugs were administered at any time during this
week; mice were only timed every other day for a full week length. In the second week
Piracetam dosages were pre-weighed at Saddleback College and placed into small test tubes, 0.5
milliliters of distilled water were dispensed into each test tube using a micropipette. Throughout
this week and the following week each group were given their corresponding amount’s every day
orally in the morning using a disposable five milliliter pipette. The mice were held by pinching
the extra skin located on their back of neck known as scruffing. Each mouse was placed back
into the maze every other day during the evening hours and timed for completion of the maze
circuit. Six sets of data were collected and were used for further analysis.
Results
The average time of the maze completion for the control group with no Piracetam collect
was 77.22 ± 5.32 seconds (Mean ± S.E.M., N=6); low dosage group fed with 5mg Piracetam
averaged 56.31 ± 6.16 seconds (Mean ± S.E.M., N=6); High dosage group with 15mg of
Piracetam averaged 46.00± 6.98 seconds (Mean ± S.E.M., N=6). A comparison graph of
completion times of maze for all groups is presented in figure 1. All sets of data were then
analyzed with ANOVA and further corrected with Bonferroni Post Hoc. ANOVA analysis
yields that there is a significance difference between all groups (p < 0.002).
Figure 1. The effects of Piracetam on cognitive performance in adult female mice to finish the maze.
High dosage of Piracetam however did significantly affect mice’s cognitive performance compare to nondosage (p < 0.002, ANOVA with Bonferroni correction). Error bars indicate ± SEM.
Using the Bonferroni post hoc correction indicates that there was not a significant
difference within the runs of the control group and low dosage experimental group (p > 0.05);
there was also no statistical difference between the runs of low dosage and high dosage
experimental groups (p > 0.05). There was, however a significant difference between the control
group and the high dosage experimental group (p < 0.05). These data are presented in table 1.
Comparison
Significant? (P <0.05?)
t
Non-dosage vs. Low dosage
No
2.388
Non-dosage vs. High dosage
Yes
3.566
Low dosage vs. High dosage
No
1.178
Table 1. Statistical comparison between three groups using Bonferroni Post Hoc correction. Post Hoc test
collected by 95% confidence level.
Discussion
This experiment demonstrated that our hypothesis was proven wrong. Although there was
a significant difference between the control group and the high dosage group, there was however
no significant difference between the low dosage group when compared to the high dosage and
control group (See Table 1). The investigators suggested that the reason why this occurred is
because the low dosage amount of (5mg/0.5mL of water) is not a sufficient dosage amount to
make a noticeable difference. On the other hand, the experimental group with the higher dosage
showed a large improvement on times and performance throughout the experiment. Concluding
that Piracetam given in larger quantities does in fact, allow the mice to complete the maze in a
faster time period due to improved memorization recall. These results were generated from
ANOVA and Bonferroni correction analysis on the mean values of maze completion time for
each group during the last two weeks (see Figure 1).
Throughout this experiment measures were taken to control the odor and scent left behind
by the mice when placed into the maze. Mice leave scent and feces as a tracking mechanism
(Bodyak, 1999). The maze was wiped down using a diluted anti-bacterial spray after each run.
Since some mice expel more waste then others some, it could have potentially caused a slight
error in time.
For further study, we propose that a follow-up experiment be performed to observe if
there are any behavioral or physical side effects on mice that could potentially cause serious
health risks. Such investigations include observing the rate of CO2 production, heart rate, and
blood pressure. Behavioral test would examine stress, anxiety, and aggression levels; which have
been seen to decrease in humans while taking Piracetam.
Acknowledgments
The researchers would like to sincerely thank Professor Teh for the guidance, assistance,
and resource preparation that made this experiment possible. The researchers would also like to
thank Saddleback College and the Department of Biological Sciences for allowing us to borrow
the equipment needed to conduct our research.
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