Effects of a Cardiotonic Medicine Danshen Pills, on Cognitive Ability and Expression of PSD-95 in a Vascular Dementia Rat Model
A widely used Chinese cardiotonic proprietary medicine, compound Danshen dripping pills (CDDP, Fufang Danshen Diwan) has also begun to be used for treatment of vascular dementia (VaD). We tried to explore the mechanism of CDDP action in this case. A VaD experimental model was built in rats by bil...
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Yanzhen Zhu Yanjun Zeng Caigu He Shu Jiang Linying Zhou Yumei Huang Changhui Qiang Yiyan Jiang 2019-02-18T10:16:51Z 2019-02-18T10:16:51Z 2016 Effects of a Cardiotonic Medicine Danshen Pills, on Cognitive Ability and Expression of PSD-95 in a Vascular Dementia Rat Model / Yanzhen Zhu, Yanjun Zeng, Caigu He, Shu Jiang, Linying Zhou, Yumei Huang, Changhui Qiang, Yiyan Jiang // Нейрофизиология. — 2016. — Т. 48, № 4. — С. 291-297. — Бібліогр.: 21 назв. — англ. 0028-2561 https://nasplib.isofts.kiev.ua/handle/123456789/148333 616.892:159.953 A widely used Chinese cardiotonic proprietary medicine, compound Danshen dripping pills (CDDP, Fufang Danshen Diwan) has also begun to be used for treatment of vascular dementia (VaD). We tried to explore the mechanism of CDDP action in this case. A VaD experimental model was built in rats by bilateral ligation of the common carotid arteries. The cognitive ability of experimental animals was evaluated in the Morris water maze test. Synaptic ultrastructural changes in the hippocampus were detected by transmission electron microscopy; expression of PSD-95 mRNA in the hippocampus was examined using hybridization in situ. The latter index (mRNA expression) in the VaD group was significantly lower than those in the CDDP and shamoperated groups (P < 0.05). CDDP treatment considerably improved disturbed ultrastructural synaptic characteristics in the hippocampus of VaD rats. The mean escape latency in the Morris water maze test was significantly shorter in CDDP-treated VaD rats, compared with that those of the VD group (P < 0.05). In the CDDP group compared to the VaD one, escape strategies improved from edge and random searches to more linear swim pathway (P < 0.05). Thus, decreasing expression of PSD-95 plays an important role in the pathogenesis of VaD. CDDP treatment improves the learning and memory ability of VaD rats by improving neural synaptic ultrastructural characteristics and increasing expression of PSD-95 mRNA in the hippocampus. Широко вживаний у Китаї патентований кардіотонічний засіб «складні пілюлі Даншен» (CDDP) почав також використовуватися для лікування васкулярної деменції (ВД). Ми досліджували можливі механізми дії цього засобу в даному аспекті. ВД моделювали у щурів, застосовуючи білатеральну перев’язку загальних сонних артерій. Когнітивні здатності експериментальних тварин оцінювали в тесті водного лабіринту Морріса. Ультраструктурні зміни синаптичних утворень у гіпокампі спостерігали, використовуючи трансмісійну електронну мікроскопію. Експресію мРНК білка PSD-95 у гіпокампі оцінювали із застосуванням методики гібридизації in situ. Останній показник (експресія мРНК) у щурів групи ВД був вірогідно нижчим, ніж у тварин контрольної групи та щурів із ВД, лікованих за допомогою CDDP. Середня затримка реакції уникання у тварин групи ВД істотно перевищувала відповідне значення в групі CDDP (P < 0.05). Стратегії уникання в останній групі були вірогідно кращими, ніж у групі ВД (збільшувалася пропорція лінійних маршрутів порівняно з «крайовими» та випадковими; P < 0.05). Зроблено висновок, що зниження експресії PSD-95 відіграє важливу роль у патогенезі ВД. Лікувальний ефект CDDP забезпечує покращення пам’яті та здатності до навчання у щурів з ВД; цей ефект опосередковується покращенням ультраструктурних показників синаптичних структур та збільшенням експресії мРНК білка PSD-95 у гіпокампі. We wish to thank H. Wu from the Animal Laboratory of the Fujian Institute of Traditional Chinese Medicine for raising the rats. This work was supported by the National Natural Science Foundation of China (grant No. 81001541) and the Natural Science Foundation of the Fujian Province (grant No. 2013J01331). en Інститут фізіології ім. О.О. Богомольця НАН України Нейрофизиология Effects of a Cardiotonic Medicine Danshen Pills, on Cognitive Ability and Expression of PSD-95 in a Vascular Dementia Rat Model Вплив кардіотонічного засобу «пілюлі Даншен» на когнітивні здатності та експресію білка PSD-95 у щурів з моделлю васкулярної деменції Article published earlier |
| institution |
Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| collection |
DSpace DC |
| title |
Effects of a Cardiotonic Medicine Danshen Pills, on Cognitive Ability and Expression of PSD-95 in a Vascular Dementia Rat Model |
| spellingShingle |
Effects of a Cardiotonic Medicine Danshen Pills, on Cognitive Ability and Expression of PSD-95 in a Vascular Dementia Rat Model Yanzhen Zhu Yanjun Zeng Caigu He Shu Jiang Linying Zhou Yumei Huang Changhui Qiang Yiyan Jiang |
| title_short |
Effects of a Cardiotonic Medicine Danshen Pills, on Cognitive Ability and Expression of PSD-95 in a Vascular Dementia Rat Model |
| title_full |
Effects of a Cardiotonic Medicine Danshen Pills, on Cognitive Ability and Expression of PSD-95 in a Vascular Dementia Rat Model |
| title_fullStr |
Effects of a Cardiotonic Medicine Danshen Pills, on Cognitive Ability and Expression of PSD-95 in a Vascular Dementia Rat Model |
| title_full_unstemmed |
Effects of a Cardiotonic Medicine Danshen Pills, on Cognitive Ability and Expression of PSD-95 in a Vascular Dementia Rat Model |
| title_sort |
effects of a cardiotonic medicine danshen pills, on cognitive ability and expression of psd-95 in a vascular dementia rat model |
| author |
Yanzhen Zhu Yanjun Zeng Caigu He Shu Jiang Linying Zhou Yumei Huang Changhui Qiang Yiyan Jiang |
| author_facet |
Yanzhen Zhu Yanjun Zeng Caigu He Shu Jiang Linying Zhou Yumei Huang Changhui Qiang Yiyan Jiang |
| publishDate |
2016 |
| language |
English |
| container_title |
Нейрофизиология |
| publisher |
Інститут фізіології ім. О.О. Богомольця НАН України |
| format |
Article |
| title_alt |
Вплив кардіотонічного засобу «пілюлі Даншен» на когнітивні здатності та експресію білка PSD-95 у щурів з моделлю васкулярної деменції |
| description |
A widely used Chinese cardiotonic proprietary medicine, compound Danshen dripping pills
(CDDP, Fufang Danshen Diwan) has also begun to be used for treatment of vascular dementia
(VaD). We tried to explore the mechanism of CDDP action in this case. A VaD experimental
model was built in rats by bilateral ligation of the common carotid arteries. The cognitive ability
of experimental animals was evaluated in the Morris water maze test. Synaptic ultrastructural
changes in the hippocampus were detected by transmission electron microscopy; expression of
PSD-95 mRNA in the hippocampus was examined using hybridization in situ. The latter index
(mRNA expression) in the VaD group was significantly lower than those in the CDDP and shamoperated groups (P < 0.05). CDDP treatment considerably improved disturbed ultrastructural
synaptic characteristics in the hippocampus of VaD rats. The mean escape latency in the Morris
water maze test was significantly shorter in CDDP-treated VaD rats, compared with that those
of the VD group (P < 0.05). In the CDDP group compared to the VaD one, escape strategies
improved from edge and random searches to more linear swim pathway (P < 0.05). Thus,
decreasing expression of PSD-95 plays an important role in the pathogenesis of VaD. CDDP
treatment improves the learning and memory ability of VaD rats by improving neural synaptic
ultrastructural characteristics and increasing expression of PSD-95 mRNA in the hippocampus.
Широко вживаний у Китаї патентований кардіотонічний засіб «складні пілюлі Даншен» (CDDP) почав також використовуватися для лікування васкулярної деменції (ВД). Ми
досліджували можливі механізми дії цього засобу в даному
аспекті. ВД моделювали у щурів, застосовуючи білатеральну перев’язку загальних сонних артерій. Когнітивні здатності експериментальних тварин оцінювали в тесті водного лабіринту Морріса. Ультраструктурні зміни синаптичних
утворень у гіпокампі спостерігали, використовуючи трансмісійну електронну мікроскопію. Експресію мРНК білка
PSD-95 у гіпокампі оцінювали із застосуванням методики
гібридизації in situ. Останній показник (експресія мРНК) у
щурів групи ВД був вірогідно нижчим, ніж у тварин контрольної групи та щурів із ВД, лікованих за допомогою
CDDP. Середня затримка реакції уникання у тварин групи
ВД істотно перевищувала відповідне значення в групі CDDP
(P < 0.05). Стратегії уникання в останній групі були вірогідно кращими, ніж у групі ВД (збільшувалася пропорція
лінійних маршрутів порівняно з «крайовими» та випадковими; P < 0.05). Зроблено висновок, що зниження експресії
PSD-95 відіграє важливу роль у патогенезі ВД. Лікувальний ефект CDDP забезпечує покращення пам’яті та здатності до навчання у щурів з ВД; цей ефект опосередковується
покращенням ультраструктурних показників синаптичних
структур та збільшенням експресії мРНК білка PSD-95 у гіпокампі.
|
| issn |
0028-2561 |
| url |
https://nasplib.isofts.kiev.ua/handle/123456789/148333 |
| citation_txt |
Effects of a Cardiotonic Medicine Danshen Pills, on Cognitive Ability and Expression of PSD-95 in a Vascular Dementia Rat Model / Yanzhen Zhu, Yanjun Zeng, Caigu He, Shu Jiang, Linying Zhou, Yumei Huang, Changhui Qiang, Yiyan Jiang // Нейрофизиология. — 2016. — Т. 48, № 4. — С. 291-297. — Бібліогр.: 21 назв. — англ. |
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NEUROPHYSIOLOGY / НЕЙРОФИЗИОЛОГИЯ.—2016.—T. 48, № 4 291
UDC 616.892:159.953
Yanzhen ZHU1, Yanjun ZENG2, Caigu HE1, Shu JIANG1, Linying ZHOU3, Yumei HUANG1, Changhui QIANG1, Yiyan JIANG1
EFFECTS OF A CARDIOTONIC MEDICINE DANSHEN PILLS, ON COGNITIVE
ABILITY AND EXPRESSION OF PSD-95 IN A VASCULAR DEMENTIA RAT MODEL
Received June 10, 2014
A widely used Chinese cardiotonic proprietary medicine, compound Danshen dripping pills
(CDDP, Fufang Danshen Diwan) has also begun to be used for treatment of vascular dementia
(VaD). We tried to explore the mechanism of CDDP action in this case. A VaD experimental
model was built in rats by bilateral ligation of the common carotid arteries. The cognitive ability
of experimental animals was evaluated in the Morris water maze test. Synaptic ultrastructural
changes in the hippocampus were detected by transmission electron microscopy; expression of
PSD-95 mRNA in the hippocampus was examined using hybridization in situ. The latter index
(mRNA expression) in the VaD group was significantly lower than those in the CDDP and sham-
operated groups (P < 0.05). CDDP treatment considerably improved disturbed ultrastructural
synaptic characteristics in the hippocampus of VaD rats. The mean escape latency in the Morris
water maze test was significantly shorter in CDDP-treated VaD rats, compared with that those
of the VD group (P < 0.05). In the CDDP group compared to the VaD one, escape strategies
improved from edge and random searches to more linear swim pathway (P < 0.05). Thus,
decreasing expression of PSD-95 plays an important role in the pathogenesis of VaD. CDDP
treatment improves the learning and memory ability of VaD rats by improving neural synaptic
ultrastructural characteristics and increasing expression of PSD-95 mRNA in the hippocampus.
Keywords: compound Danshen dripping pill, hippocampus, Morris water maze,
PSD-95, synapsis, vascular dementia.
1 Department of Integrative Medicine, Fujian University of Traditional
Chinese Medicine, Fujian, China.
2 Biomechanical and Medical Information Institute, Beijing University of
Technology, Beijing, China.
3 Department of Electron Microscopy, Fujian Medical University, Fujian,
China.
Correspondence should be addressed to Yanjun Zeng
(e-mail: yjzeng@bjut.edu.cn).
INTRODUCTION
Vascular dementia (VaD) is a clinical syndrome of
cognitive decline caused by ischemic, hemorrhagic,
or oligemic injury to the brain as a consequence of
cardiovascular and/or cerebrovascular diseases [1]. In
the United States and Europe, it accounts for 10-20%
of dementia cases [2, 3] and greatly impacts on the
patients’ life quality and national economies. Within an
aging population, VaD will become the more prevalent
disease in the future. Officially approved definitive
medical or surgical treatments for vascular dementia still
remain limited [4]; thus, the development of effective
treatments for VaD would be of great importance.
Compound Danshen dripping pills (CDDP; Fufang
Danshen Diwan in Chinese) is a well-known Chinese
proprietary medicinal formula. It is exclusively broadly
used in China and some other countries for treating
vascular diseases, including cardiovascular and
cerebrovascular ones [5, 6]. Clinically, CDDP have been
shown to produce beneficial effects on VaD and stroke
patients in China. Our previous experiments showed
that CDDP noticeably improved the learning and
memory ability of VaD rats by inhibiting expression of
Bax and caspase-3 in the hippocampus. These evidences
suggest a significant positive effect of CDDP on VaD.
However, the molecular mechanisms of such effect
remained poorly known, and this topic is worth further
study.
There is a widespread agreement that memory can
be encoded by activity-dependent changes in the
transmission strength of synapses. Considerable data
showed that synaptic ultrastructural and functional
impairment contributes to VaD [7]. One of important
postsynaptic density-related proteins is PSD-95.
It plays a central role in the regulation of synaptic
functions, and it is also significant for the learning and
memory ability [8]. At the same time, the role of PSD-
95 in the pathogenesis of VaD remained unclear. The
NEUROPHYSIOLOGY / НЕЙРОФИЗИОЛОГИЯ.—2016.—T. 48, № 4292
Yanzhen ZHU, Yanjun ZENG, Caigu HE, et al.
aim of this our study was to evaluate the role of PSD-
95 in the VaD pathogenesis and to estimate the effect
of CDDP on the expression of PSD-95 in VaD rats.
METHODS
Animal Selection. A total of 40 healthy female
Sprague–Dawley rats, belonging to a specific
pathogen-free grade, aged 12 months, and weighing
432 ± 30 g, were provided by the Research Center of
Laboratory Animals (Academy of Military Medical
Sciences, China, permission No. SCXK (Jun) 2007-
004). Animals were maintained under conditions of
the controlled temperature (22 ± 2°C), humidity (50-
70%), and illumination (12-h light-dark cycle with
lights on at 7:00) with free access to food and water.
Thirty-seven rats with good learning and memory
abilities were selected according to the Morris water
maze criteria, while three animals were eliminated.
Preparation and Grouping of Animal Models. Nine
rats were selected to the sham-operated (sham) group,
and 28 rats were used for VaD modeling, which was
induced by bilateral occlusion of the common carotid
arteries [9]. Under anesthesia (choralhydrate 3 mg/kg),
rats were fixed in a supine position, and a midline
incision of the cervical skin was made to expose
bilaterally the carotid arteries; the latter were ligated
with double silk sutures; then the rats were regularly
fed and observed. After induction of the VaD model,
the surviving rats were randomly assigned into the VD
(n = 10) and CDDP-treated (n = 10) groups. The sham
group received the same surgical procedures, but the
common carotid arteries were not occluded.
CDDP Used to Treat the VaD Rats. Ten days after
surgery, CDDP (Tasly Pharmaceutical Corporation,
Tanjian, China) was dissolved in normal saline and
administered to rats of the CDDP group. Each rat
this group was given CDDP at dose of 75 mg/kg by
gastrogavage once a day for 20 days. The rats of the
sham and VaD groups were reared under the same
conditions, without any treatment.
Morris Water Maze Test. Two months after
surgery, the behavioral Morris water maze test was
conducted. A circular tank (150 cm diameter, 50 cm
deep) was filled with 17-cm deep water of 22-26°C.
Four entering points were marked on the wall of the
pool, and the pool was divided into four quadrants.
The platform (12.5 cm diameter, 15 cm high) was
placed in the third quadrant; it was submerged 2 cm
below the water surface [9].
The Navigation Test for Evaluation of the
Learning and Memory Ability. The test lasted for
5 days with four trials per day. Each trial began by
releasing the rat into water, with its muzzle toward the
pool wall at one of the four placement points. The time
to find the platform within a 2-min limit was recorded.
If the rat failed to find the platform within 2 min, the
experimenter moved the animal to the platform, where
it remained for 10 sec, and the escape latency was
measured within 2-min-long interval.
Spatial Memory Ability of Rats Analyzed by the
Spatial Probe Test. The platform was removed from the
pool, and rats were allowed to search for the platform
within 120 sec. Swim paths were recorded to determine
search strategies; this included the search along an edge
swim path (edge type), that along a random swim path
(random type), a trend toward a straight swim path
(trend type), and a linear swim path (line type).
Electron Microscopy of the Hippocampus. After
cervical dislocation，the animals were sacrificed, the
rat brain was quickly taken off, and the hippocampus
was isolated on an ice plate. Hippocampal tissues were
kept in a fixative containing 3% glutaraldehyde and
1.5% paraformaldehyde (in 0.1 M PBS, pH 7.4). A tis-
sue block (about 1 mm ×1 mm × 3 mm) of the CA1 re-
gion was taken from the hippocampus and fixed in the
same fixative for 24 h. Then it was immersed in the
fixative including 1% osmic acid and 1.5% potassium
ferrocyanate for 1.5 h. The tissue was stained over-
night with uranyl acetate, dehydrated with ethanol and
acetone, and embedded in Epon 618. One-µm-thick
sections were taken from each block and stained with
uranyl acetate and lead citrate. Ultrathin sections were
cut, placed on uncoated or coated grids, and observed
with a Philips - EM208 electron microscope.
Preparation of Paraffin Sections. Under anesthesia
with choral hydrate, the rat brain was taken off, and
hippocampal tissues were isolated on an ice plate,
postfixed in 10% neutral formalin for 12 h, dehydrated
through an ethanol series, embedded in paraffin, and
serially sectioned (6-μm thick coronal sections).
In Situ Hybridization Used to Evaluate
Hippocampal PSD95 mRNA Expression. Hybridi za-
tion in situ was performed according to the instruction
for a kit of hybridization in situ given by the producer
(Boshide, China). Briefly, after deparaffinage and
rehydration, the brain section was kept in 3% H2O2,
at room temperature for 5-10 min, to inactivate the
endogenous enzyme, and then digested with peptase
for 2 min. After processing with 20 µl prehybridization
solution, each section was processed in 20 µl
NEUROPHYSIOLOGY / НЕЙРОФИЗИОЛОГИЯ.—2016.—T. 48, № 4 293
EFFECTS OF A CARDIOTONIC MEDICINE DANSHEN PILLS
hybridization solution overnight at 40℃, in biotinylated
mouse anti-digoxin at 37℃ for 1 h, and in biotinylated
peroxidase at 37℃ for 20 min. With horseradish
peroxidase cata lys is , 3 ,3N-diaminobenzidine
tertrahydrochloride (DAB) was used to color a
masculine position of the section. PSD-95 mRNA-
positive cells and nerve cells in the hippocampal CA1
region were observed using a Nikon 55i microscope.
The percentage of PSD-95 mRNA-positive cells in each
hippocampal CA1 section was calculated.
Statistical Analysis. Data were statistically treated
by the first coauthor using SPSS 13.0 software. Data
for the escape latency and results of hybridization in
situ were analyzed with one-way ANOVA and the LSD
post hoc test. The χ2 test and partition χ2 tests were
used for spatial probe analysis.
RESULTS
Learning and Memory Ability of VaD Rats Treated
with CDDP. The navigation test in the Morris water
maze was used to evaluate the learning and memory
ability of the examined rats. The mean escape latencies
in the sham, VaD, and CDDP groups were 10.72 ±
2.40, 43.00 ± 5.38 and 19.71 ± 4.28 sec, respectively.
The LSD test showed that the average latency in the
VaD group was significantly longer than those in
the sham and CDDP groups (P < 0.05). The χ2 test
showed that escape strategies in the spatial probe test
showed significant differences among the three groups
(P < 0.05), and partition by the χ2 test demonstrated
noticeable differences of escape strategies between
the VaD group and CDDP groups. Escape strategies
were clearly improved from edge and random searches
to more linear swim pathways in the CDDP group,
compared to those in the VaD group (Fig. 1).
F i g. 1. Proportions, %, of using different spatial strategies (1-4)
in the water maze test by rats of the experimental groups. 1-4) Line
type, 2) trend type, 3) random type, and 4) edge type.
Р и с. 1. Нормована частота (%) використання різних просто-
рових стратегій у тесті водного лабіринту щурами експеримен-
тальних груп.
F i g. 2. Representative examples of ultrastructural characteristics
of synaptic structures in the hippocampus of rats of the sham, VaD,
and CDDP groups (A–C, respectively).
Р и с. 2. Типові приклади ультраструктури синаптичних утво-
рень у гіпокампі щурів груп контролю, VaD та CDDP (А–С
відповідно).
50
%
A
B
C1 1 12 2 2
sham VaD
300 nm
300 nm
300 nm
CDDP
3 3 34 4 4
40
30
20
10
0
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Yanzhen ZHU, Yanjun ZENG, Caigu HE, et al.
F i g. 3. PSD-95 mRNA-positive cells (hybridization in situ) in rats
of the sham, VaD, and CDDP groups (A-C, respectively).
Р и с. 3. PSD-95 мРНК-позитивні клітини (виєвлення за допо-
могою гибридізації in situ) у щурів трьох груп (АС).
Synaptic Ultrastructural Patterns in the
Experimental Groups. In rats of the sham group; the
presynaptic membranes, postsynaptic membranes, and
synaptic cleft were clear. There were many synaptic
vesicles in the presynaptic components, and prominent
postsynaptic densities (PSDs) were observed in the
postsynaptic components.
In the VaD group; the presynaptic and postsynaptic
membranes got unclear. Synapses had noticeably fewer
synaptic vesicles in their presynaptic components and
thinner postsynaptic densities in the postsynaptic
components. Clear swelling appeared in some
presynaptic and postsynaptic components.
As was found in the CDDP group, the treatment with
this mean clearly improved synaptic ultrastructural
patterns in the hippocampus. Compared with the
VaD group, the presynaptic and postsynaptic surfaces
got clearer, much more synaptic vesicles were
observed, and the postsynaptic densities became
thicker. Swelling in the presynaptic and postsynaptic
components alleviated.
Expression of PSD 95 mRNA Examined by
Hybridization in Situ. PSD-95 is predominantly
localized within synapses, and it was shown to play
an important role in synaptic plasticity. Hybridization
in situ was conducted to assess the role of PSD-95 in
CDDP treating of VaD. PSD-95 mRNA-positive loci
F i g. 4. Percentage of PSD 95 mRNA-positive cells in slices of the
hippocampal CA1 region.
*P < 0.05 vs. the sham group; +P < 0.05 vs. the VaD group. Data
are expressed as means ± s.d. for each group. Other designations are
similar to those in Fig. 1.
Р и с. 4. Нормована кількість (%) клітин позитивних щодо
мРНК PSD 95, у зрізах зони СА1 гіпокампа.
A
B
C
50 μm
50 μm
50 μm
100
80
60
40
20
0
%
sham VaD CDDP
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EFFECTS OF A CARDIOTONIC MEDICINE DANSHEN PILLS
appeared in the cytoplasm and nuclei of hippocampal
CA1 nerve cells (Fig. 3). Compared with that in
the sham group, expression of PSD 95 mRNA was
significantly lower in the hippocampus of VaD-
group rats (P < 0.05). CDDP treatment considerably
increased expression of PSD 95 mRNA, as compared
to that in VaD rats (P < 0.05) (Figs. 3 and 4).
DISCUSSION
The Morris water maze test gas been widely used to
evaluate the learning and memory ability (especially
spatial memorization) in rats [10]. The mean escape
latency refers to the average time for rats to initiate
finding of the platform, which reflects the level of
learning and memory ability of the animals. Search
strategy in the spatial probe test refers to the path of
rats to swim to the safe space (the platform) according
to the surrounding environment, and the respective
indices reflect the space memory ability of rats. This
mainly includes the edge, random, tend, and straight
types. Navigation test results showed that the average
escape latency in the VaD group was significantly
longer than that in normal rats, and CDDP treatment
noticeably decreased the average escape latency in
VaD rats. The spatial probe test results proved that the
proportions of edge-type and random-type strategies
were noticeably greater in the VaD group compared
with those in the sham group. This was indicative of
the poor space memory ability in the VaD group [11,
12]. It was obvious that CDDP treatment improved the
search strategies of VaD rats. A complex of these data
indicated that there is a significant positive effect of
CDDP on the cognitive function in the above rats.
The PSD is a specialized cytoskeletal structure
lying beneath the postsynaptic membrane. It is
involved in the regulation of synaptic functions and
modulation of postsynaptic responses [13]. PSD-95
is the best-characterized protein member of the PSD
family and an effective postsynaptic biomarker. It
enhances postsynaptic clustering and plays important
roles in integration of postsynaptic signals [14]. In
ischemic rats, PSD-95 expression in the hippocampus
was significantly lower compared with that in sham-
operated animals [15, 16]. This fact suggests that
down-regulation of PSD-95 is an important factor
in ischemic encephalopathy. Our study showed that
ultrastructural impairment of the synapses including
thinning of the PSD was obvious in the VaD group, and
hippocampal PSD-95 mRNA expression significantly
decreased in this group. Such results suggest that
hippocampal ischemia induced by bilateral switching-
off of the common carotid arteries led to obvious
reduction of the postsynaptic density thickness and a
decrease in the expression of PSD-95 mRNA. It also
affected the efficiency of synaptic transmission. These
events, naturally, resulted in a learning and memory
deficits.
CDDP is a classical and widespread prescription
for the treatment of cardiovascular diseases. Recently,
it began also to be used for that of cerebrovascular
disease [17]. Clinically, CDDP exerts significant
effects in treating VaD by slowing-down progression
of this pathology. The reason may be related to the
CDDP function of promoting blood circulation
and removing blood stasis. The main constituents
of CDDP are radix salviae miltiorrhizae, radix
notoginseng, and borneolum syntheticum. Radix
salviae miltiorrhizae is the most important component
of this medicine. It exerts multifacial positive
effects (decreasing constriction of the blood vessels,
improving circulation, relieving blood stasis, and
changing the blood viscidity). In ischemia rats,
active constituents of radix salviae miltiorrhizae
provide cerebroprotection by reducing apotposis
and inhibiting NOS gene expression [18, 19]. Radix
notoginseng has the following effects: decreasing
platelet superficial activation, inhibiting platelet
adhesion and aggregation, preventing thrombosis, and
improving microcirculation [20]. Borneolum increases
the permeability of the blood-brain barrier [21].
Our experiments convincingly demonstrated
that CDDP improves cognitive ability of VaD rats,
positively modifies injured synaptic structures in
VaD rats, and promotes PSD-95 mRNA expression in
the cerebral structure crucially important for spatial
memory (hippocampus).
Thus , our resul ts showed that decreased
expression of PSD-95 probably plays a key role in
the pathogenesis of VaD, and that CDDP exerts a
protective influence on the synaptic plasticity in the
hippocampus by improving expression of mRNA of
this synaptic protein. Our study provides behavioral
and biochemical evidences for a possibility of using
CDDP therapy in VaD. However, the mechanism of
how cerebral ischemia leads to decreased expression
of PSD-95 remains unclear, and the mechanism of how
CDDP affects the hippocampal neuronal mechanisms
requires further studies.
NEUROPHYSIOLOGY / НЕЙРОФИЗИОЛОГИЯ.—2016.—T. 48, № 4296
Yanzhen ZHU, Yanjun ZENG, Caigu HE, et al.
Acknowledgments. We wish to thank H. Wu from the
Animal Laboratory of the Fujian Institute of Traditional
Chinese Medicine for raising the rats. This work was supported
by the National Natural Science Foundation of China (grant
No. 81001541) and the Natural Science Foundation of the
Fujian Province (grant No. 2013J01331).
Rats were handled according to the National Guidelines
of China for the Care and Use of Animals for Scientific
Purposes. Protocols were approved by the Animal Care and
Use Committee at the Fujian University of Traditional Chinese
Medicine.
The authors, Yanzhen Zhu, Yanjun Zeng, Caigu He, Shu Jiang,
Linying Zhou, Yumei Huang, Changhui Qiang, Yiyan Jiang,
confirm that they have no conflicts of any kind related to the
commercial or financial problems, relations with organizations
or persons, which could in any way be associated with the
investigation, and with the relationship of the co-authors of the
article.
Я. Жу1, Я. Зен2, Л. Хе1, Ш. Джян1, Л. Жоу3, Ю. Хуан1,
Ч. К’ян1, Йї. Джян1
ВПЛИВ КАРДІОТОНІЧНОГО ЗАСОБУ «ПІЛЮЛІ ДАН-
ШЕН» НА КОГНІТИВНІ ЗДАТНОСТІ ТА ЕКСПРЕСІЮ
БІЛКА PSD-95 У ЩУРІВ З МОДЕЛЛЮ ВАСКУЛЯРНОЇ
ДЕМЕНЦІЇ
1 Університет традиційної китайської медицини, Фуджянь
(Китай).
2 Інститут біомеханіки та медичної інформації Пекінського
технологічного університету (Китай).
3 Фунджянський медичний університет (Китай).
Р е з ю м е
Широко вживаний у Китаї патентований кардіотонічний за-
сіб «складні пілюлі Даншен» (CDDP) почав також вико-
ристовуватися для лікування васкулярної деменції (ВД). Ми
досліджували можливі механізми дії цього засобу в даному
аспекті. ВД моделювали у щурів, застосовуючи білатераль-
ну перев’язку загальних сонних артерій. Когнітивні здат-
ності експериментальних тварин оцінювали в тесті водно-
го лабіринту Морріса. Ультраструктурні зміни синаптичних
утворень у гіпокампі спостерігали, використовуючи транс-
місійну електронну мікроскопію. Експресію мРНК білка
PSD-95 у гіпокампі оцінювали із застосуванням методики
гібридизації in situ. Останній показник (експресія мРНК) у
щурів групи ВД був вірогідно нижчим, ніж у тварин конт-
рольної групи та щурів із ВД, лікованих за допомогою
CDDP. Середня затримка реакції уникання у тварин групи
ВД істотно перевищувала відповідне значення в групі CDDP
(P < 0.05). Стратегії уникання в останній групі були віро-
гідно кращими, ніж у групі ВД (збільшувалася пропорція
лінійних маршрутів порівняно з «крайовими» та випадко-
вими; P < 0.05). Зроблено висновок, що зниження експресії
PSD-95 відіграє важливу роль у патогенезі ВД. Лікуваль-
ний ефект CDDP забезпечує покращення пам’яті та здатнос-
ті до навчання у щурів з ВД; цей ефект опосередковується
покращенням ультраструктурних показників синаптичних
структур та збільшенням експресії мРНК білка PSD-95 у гі-
покампі.
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