Baoshun, Hao

Bin, Zhou

Dinghui, Liu

Lin, Chen

Lin, Wu

Min, Wang

Shujie, Yu

Xiaoxian, Qian

Yong, Liu

Zhiming, Song

Elsevier - Publisher Connector 

Results: Results showed that treatment with AVE 0991 could reduce AngII-induced
proliferation of cardiac ﬁbroblasts, manifesting decreased DNA synthesis and collagen
production. Moreover, treatment with AVE 0991 could attenuate AngII-induced
activation of Smad signaling pathways. Furthermore, the beneﬁcial effects and Smad
pathways’s changes of AVE0991 were abolished by A-779.
Conclusions: Our data showed that AVE 0991 attenuated proliferation of cardiac
ﬁbroblast, which may be due to the inhibition of Smad pathways.
GW25-e1651
Identiﬁcation of Stim1 as a Candidate Gene for Exaggerated Sympathetic
Response to Stress in the Stroke-Prone Spontaneously Hypertensive Rat
Xiao Bing1,2, Liu Fan1, Ohara Hiroki2, Nabika Toru2
1Department of Cardiology, The Second Hospital of Hebei Medical University,
Shijiazhuang, Hebei province, China, 2Department of Functional Pathology, Shimane
University School of Medicine, Izumo, Japan
Objectives: The stroke-prone spontaneously hypertensive rat (SHRSP) is known to
have exaggerated sympathetic nerve activity to various types of stress, which might
contribute to the pathogenesis of severe hypertension and stroke observed in this
strain. Previously, by using a congenic strain (called SPwch1.72) constructed between
SHRSP and the normotensive Wistar-Kyoto rat (WKY), we showed that a 1.8-Mbp
fragment on chromosome 1 (Chr1) of SHRSP harbored the responsible gene (s) for the
exaggerated sympathetic response to stress. To further narrow down the candidate
region, in this study, another congenic strain (SPwch1.71) harboring a smaller frag-
ment on Chr1 including two functional candidate genes, Phox2a and Ship2, was
generated.
Methods: Sympathetic response to cold and restraint stress was compared among
SHRSP, SPwch1.71, SPwch1.72 and WKY by three different methods (urinary
norepinephrine excretion, blood pressure measurement by the telemetry system and
the power spectral analysis on heart rate variability).
Results: The results indicated that the response in SPwch1.71 did not signiﬁcantly
differ from that in SHRSP, excluding Phox2a and Ship2 from the candidate genes. As
the stress response in SPwch1.72 was signiﬁcantly less than that in SHRSP, it was
concluded that the 1.2-Mbp congenic region covered by SPwch1.72 (and not by
SPwch1.71) was responsible for the sympathetic stress response. The sequence
analysis of 12 potential candidate genes in this region in WKY/Izm and SHRSP/Izm
identiﬁed a nonsense mutation in the stromal interaction molecule 1 (Stim1) gene of
SHRSP/Izm which was shared among 4 substrains of SHRSP. A western blot analysis
conﬁrmed a truncated form of STIM1 in SHRSP/Izm. In addition, the analysis
revealed that the protein level of STIM1 in the brainstem of SHRSP/Izm was
signiﬁcantly lower when compared with WKY/Izm.
Conclusions: Our results suggested that Stim1 is a strong candidate gene responsible
for the exaggerated sympathetic response to stress in SHRSP.
GW25-e1653
SIRT1 enhances Therapeutic Efﬁcacy of Aged Mesenchymal Stem Cells in Rat
Myocardial Infarction via lightening MSCs Aging and heightening Stress
Resistance
Huiqiang Chen, Jian-an Wang
Cardiovascular Key Lab of Zhejiang Province, the Second Afﬁliated Hospital, School
of Medicine, Zhejiang University
Objectives:Mesenchymal stem cells (MSCs) hold great promise for stem cell therapy
and are gradually used in clinical trials in recent years. However, advanced age is a
negative factor for autologous MSCs transplantation.
Methods: For in vitro study, we evaluated cell senescence using SA-B-gal method
while for evalulated the stress response we conducted H2O2-induced hypoxia/
ischemia model in MSCs. The conditioned medium was collected and was added to
HUVEC to observe the alteration of tube-formation between vectot-aged MSCs and
SIRT1-aged MSCs. For experiments in vivo, we ﬁrst conducted ischemic female heart
model induced by left anterior descending (LAD) ligation. At day 3 after male cell
transplantation, the cell survival was evaluated by SRY detection. At day 28 after cell
transplantation, heart function and ﬁbrosis were analyzed by ECHO and Masson
staining respectively. Morover, vascular density was evaluatied by CD31 and SMA
staining.
Results: In the present study, we evaluated the effect of SIRT1 overexpression in
MSCs from aged rat in vivo and in vitro. Compared with vector-aged MSCs, SIRT1
modiﬁed-aged MSCs signiﬁcantly promote cellular survival in the ischemic heart
induced by left anterior descending ligation at day 3 after cell transplantation.
Moreover, SIRT1 overexpression in aged MSCs could down-regulate ﬁbrosis and
up-regulate vascular density, consequently contribute to the improvement of heart
function. In vitro data demonstrated that forced expression of SIRT1 inaged MSCs
could contribute to signiﬁcantly attenuation of MSCs senescence and the aug-
mentation of MSCs survival under hypoxia/ischemia condition. The improvement of
angiogenesis was also observed in vitro when SIRT1 was overexpressed in aged
MSCs compared with vector-aged MSCs, which might be attributed to the up-
regulation of paracrine factors including AngI and bFGF.
Conclusions: Together, our data support the notion that forced expression of SIRT1
inaged MSCs could ameliorate the phenotypes and function of aged MSCs, conse-
quently contribute to the improvement of cell-based therapy for myocardial infarction.C76 JACC Vol 64/16/Suppl C j OctobGW25-e2290
Effect of Sini Decoction on the expression of SOD and MDA in EAhy926 cells
injured by homocysteine
Liu Yong, Song Zhiming, Zhou Bin, Yu Shujie, Liu Dinghui, Hao Baoshun, Wu Lin,
Wang Min, Chen Lin, Qian Xiaoxian
Department of Cardiology, The Third Afﬁliated Hospital of Sun Yat-sen University
Objectives: To detect the effect of Sini Decoction on the expression of SOD and
MDA in EAhy926 cell injured by homocysteine.
Methods: Model of EAhy926 cell injured by homocysteine was made, the protection
on the EAhy926 cell of Sini Decoction with different dosages were observed. SOD
and MDA concentration of cell culture ﬂuid was detected, effect of Sini Decoction on
the expression of protein of SOD1 in EAhy926 cells were observed by Western-blot,
and effect of Sini Decoction on the expression of mRNA of SOD1 in EAhy926 cell
were observed by ﬂuorescent quantitation PCR.
Results: After Model of EAhy926 cell injured by homocysteine was made, we found
that cultured with 0.5, 1.0, 2.0, 4.0, 8.0mmol/L homocysteine, cells grew less than
cultured with normal culture medium, as culturing with homocysteine 4.0mmol/L for
24h did lower damage to cells and could induce effective cell injuring, it was made to be
the model of injury. To detect the effect of Sini Decoction on EAhy926 cell injured by
homocysteine, well growing EAhy926 cells were cultured in culture plate. 24h later,
cells were cultured with medium containing Sini Decoction 0, 0.25, 0.5, 1.0g/ml
respectively for 30 minutes, then cultured with medium containing homocysteine
4.0mmol/L for 24h. It was found that, compared with control group, attached cells in Sini
Decoction groups grew better, and attached cells in Sini Decoction 1.0g/ml plus
homocysteine 4.0mmol/L group grew best. It was found that compared with control
group, there was no obvious change of SOD (5.77650.1602 vs 5.6087±0.661u/ml)
and MDA (0.6667±0.1649 vs 0.6775±0.1528 nmol/ml) concentration of cell culture
ﬂuid in Sini Decoction 1.0 g/ml group, but in homocysteine 4.0mmol/L medol group,
SOD (2.6434±0.5765 u/ml) concentration of cell culture ﬂuid decreased oberviously
and MDA (1.2381±0.1649 nmol/ml) concentration of cell culture ﬂuid increased
oberviously, while in Sini Decoction groups, SOD concentration of cell culture
ﬂuid increased and MDA concentration of cell culture ﬂuid decreased, in Sini
Decoction 1.0g/ml plus homocysteine 4.0mmol/L group it was the most obvious
(SOD4.9599±0.5884 u/ml, MDA0.7143±0.1428 nmol/ml, P<0.05). Detected by
Western-blot, it was found that, campared with control group, there was no obervious
change of protein of SOD1 in Sini Decoction 1.0 g/ml group, but in homocysteine
4.0mmol/L model group, expression of SOD1 protein weakened obviously, and in Sini
Decoction groups, expression of SOD1 protein enhanced, and in Sini Decoction 1.0g/ml
plus homocysteine 4.0mmol/L group it was the most obvious (P<0.05). Detected by
ﬂuorescent quantitation, it was found that, compared with control group, there was no
obvious change of mRNA of SOD1 in Sini Decoction 1.0g/ml group, but in homo-
cysteine 4.0mmol/Lmodel group, expression of SOD1mRNAweakened obviously, and
in Sini Decoction groups, expression of SOD1 mRNA enhanced, and in Sini Decoction
1.0g/ml Plus homocysteine 4.0mmol/L group, it was the most obvious (P<0.05).
Conclusions: Homocysteine may injure EAhy926 cell by suppressing the expression
of SOD, while Sini Decoction may protect EAhy926 cell by enhancing the expression
of SOD.
GW25-e2345
MiR-211 mediated hypoxia induce bone marrow mesenchymal stem cells
migration through STAT5A
Hu Xinyang, Jian-an Wang
Cardiovascular Key Lab of Zhejiang Province, the Second Afﬁliated Hospital, School
of Medicine, Zhejiang University
Objectives: Efﬁcacy of intravenous administration of mesenchymal stem cells
(MSCs) for myocardial infarction (MI) is limited by low cell migration to the damaged
myocardium. Our previous study demostrated that migration ability of MSCs
enhanced by hypoxia preconditioning (HPC). miRNA microarray displayed that miR-
211 exhibited the most signiﬁcant change between HPC and normoxia cultured
MSCs. The aim of this study is to identify whether miR-211 regulates MSCs
migration and the potential mechanism.
Methods: In vitro, transwell assay were used to assess the migration ability of MSC
moduated by miR-211 using miR-211 overexpressing and knockdown lentivirus. The
target gene of miR-211 predicted by Targetscan, which were veriﬁed by PCR, western
blot and lucifurase assay. We knocked down the target gene and assessed the
migration capability again. Chromatin immunoprecitation (ChiP) were used to explore
the transcription factors that regulate the expression of miR-211. To evaluate the effect
of miR-211 on MSCs migration in vivo, miR-211-mimic and miR-211-shRNA MSCs
from male mice were intravenously delivered 24h after MI, the engraft cells were
detected by RT-PCR of SRY gene.
Results: Quatitative RT-PCR showed that miR-211 expression of MSCs upregulated
by HPC. MiR-211 mimic improved MSCs migration by 31.03% (P<0.05), however,
knockdown miR-211 using shRNA attenuated MSCs migration ability signiﬁcantly.
Signal transducer and activator of transcription 5A (STAT5A) was predicted as one of
target genes of miR-211, and was conﬁrmed by PCR and Western blot, it showed
miR-211 overexpression dramatically decreased STAT5A expression, while miR-211
knockdown upregulated STAT5A. The luciferase assay showed the similar results.
Functional experiment showed that STAT5A knockdown reverse the inhibition of
MSCs migration induced by miR-211-shRNA. Intrestingly, ChiP assay showed thater 16–19, 2014 j GW-ICC Abstracts/Basic and Translational Medicine


GW25-e2290 Effect of Sini Decoction on the expression of SOD and MDA in EAhy926 cells injured by homocysteine 

file:///data/core-remote/dit/data/elsevier/pdf/e04/aHR0cDovL2FwaS5lbHNldmllci5jb20vY29udGVudC9hcnRpY2xlL3BpaS9zMDczNTEwOTcxNDAzMjUwMQ==.pdf

GW25-e2290 Effect of Sini Decoction on the expression of SOD and MDA in EAhy926 cells injured by homocysteine

Abstract

Similar works

Full text

Available Versions

Elsevier - Publisher Connector