2. Introduction
AD has become one of the diseases that seriously affect the health of the elderly.
The most important pathologic hallmarks in AD are neurofibrillary tangles and
amyloid plaques.
Excessive metal elements can aggravate dementia symptoms.
ROS can act on DNA fragments to cause DNA breakage and base oxidation,
thereby aggravating nerve damage.
Glycogen synthase kinase-3 (GSK-3), a key role in the pathogenesis
of Alzheimer’s disease (AD), has been linked.
PIMPC: metal-chelating imine GSK-3 inhibitor.
In the study, Aβ/Cu2+-induced AD rat model was established and
treated with PIMPC.
PIMPC can´t only down-regulate the high expression levels of Aβ, tau
and p-tau proteins, but also chelate Cu and aluminum.
3. Study on multi-target effects of
PIMPC on Aβ/Cu2+-induced
Alzheimer’s disease model of rats
General Objective
4. 1.RNA of hippocampus was
extracted with trizol (50mg)
reagent
2. Centrifugation (10min),
supernatant was transferred with
chloroform.
3. Supernatant + Isopropanol
= RNA precipitate
4. RNA precipitate + Ethanol.
Centrifuged and evaporated to
dryness.
5. Dissolved in amount of RNase-
free water
6.Reverse transcription: RNA
solution to a PCR tube containing
Oligo and Random Hexamer Prime
7.Solution incubated at 65 degrees
in PCR amplifier and then cooled
on ice.
8. Remaining solvent added to the
tube placed at 42 degrees (1h) , and
then incubated at 70 degrees.
9. RTPCR reaction system. Copy
number of the gene was
calculated.
Methods
Real-time quantitative PCR analysis
5. Methods
Western blotting
Hippocampal tissue was added with protease inhibitors to
form a homogenate which was then lysed on ice for 30 minutes
and centrifuged to obtain protein solution, determined by BCA
Kit.
The protein sample was separated by electrophoresis (SDS-
PAGE), to be subsequently transferred. Milk was used to avoid
absorption of antibodies. Membranes were incubated with
antibodies for Aβ tau, phospho-tau (p-tau) and Bcl-2.
Signals were determined by an enhanced chemiluminescence
method, and the film was scanned for gray-scale value
analysis.
Hematoxylin and eosin staining and terminal dUTP nick
end labelling (TUNEL) assay
Two randomly selected rats in each group were anesthetized by
injection of 10% chloral hydrate after 24 hours of fasting. Brain
tissue was removed and fixed in 4% paraformaldehyde solution
for 24 hours.
Brain tissues were dehydrated in ethanol solutions and washed
with xylene and cut into sections. Sections were stained with
hematoxylin and eosin for observation. Remaining sections
were used for TUNEL assay.
Passed through another series of processes, three visual fields
were randomly selected for cell counting under a fluorescence
microscope.
Determination of ATPase, antioxidant indexes, AChE,
NOS and antioxidant indexes
After a 24-h fast, decapitated blood from the rats was
centrifuged to obtain serum, brain tissue, and hippocampus was
separated. The brain tissue was added in precooled saline to
make a 10% tissue homogenate. Homogenates were centrifuged
and supernatants were separated for analysis.
According to the kit instructions, total protein and MDA
concentrations and MDA activities were determined.
Total protein and MDA and ATPase, SOD, GSH-Px, AChE and NOS
activities in the brain were determined.
6. Results
Alpha Beta Myeloid expression at high doses of
PIMPC was decreased.
Tau and P tau are decreased in the low and high
dose PIMPC group making the comparison with
Copper for example.
No significant changes in Bcl-2, Bax and Caspase
expression.
Group AB/Cu2+ hippocampal cleaved caspase was
slightly elevated compared to control.
High and low dose PIMPC groups had decreased
expression of cleaved caspase.
Note: A, control group; B, Aβ/Cu2+ group; C,
donepezil group; D, low-dose PIMPC group; E, high-
dose PIMPC group.
7. Results
Considering the CONTROL image we observe them
in order and narrow.
When compared with the presence of Alpha Beta
Miloid or Copper the cellular vacuoles start to
appear.
The change with drug and PIMPC the distribution
returns to acquire a narrow order and clearly a
reduction of the vacuoles.
8. Results
It is observed how apoptosis was generated
in hippocampal cells of the Alpha Beta/Cu2+
group.
Therefore, the rate of apoptosis in
hippocampal neurons is seen to be higher
compared to the control image.
Again, the presence of drug and PIMPC were
significantly lower.
10. Conclusions
The genetic study, there are many diseases
that thanks to this contribution have been
benefited at the time of granting a diagnosis
or search for the cause of the pathology from
the innermost part with the aspiration of
finding a good treatment.
In relation to the article, you can clearly see
the point I made above. The study of a
component such as PIMPC with the respective
tests has given the green light to the
prevention or improvement of the quality of
life of individuals with Alzheimer's disease,
Molecular biology has proven to be an indispensable discipline in the medical
field. There are several reasons, but in my personal opinion, the following stand
out:
