2015 Conferences
2015 RRSH 9th international meeting on Rapid Responses to Steroid Hormones
The First Joint Annual Conference of Health Science & The First Cross-Strait Symposium on Endocrinology and Metabolism & The Tenth Chiang Shou-Teh Symposium
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S01
Renoprotection of Renin-Angiotensin-Aldosterone Blockade in Chronic Kidney Disease Patients with Hypertension
Der-Cherng Tarng
Department and Institute of Physiology, National Yang-Ming University, and Division of Nephrology, Department of Medicine, Taipei Veterans General Hospital Taipei, Taiwan
dctarng@vghtpe.gov.tw
The benefit of using a renin-angiotensin-aldosterone system blocker such
as an angiotensin-converting en- zyme inhibitor (ACEI) or an angiotensin II
receptor blocker (ARB) for patients with advanced chronic kidney dis- ease
(CKD) remains undetermined. We have conducted a prospective cohort study to assess the effectiveness and safety of ACEI/ARB use for the advanced CKD 5 patients with
hypertension and anemia. From January 1, 2000, through June 30, 2009, we
selected 28,497 hypertensive adult patients with CKD. Serum creatinine levels were greater than 6 mg/dl,
hematocrit levels were less than 28%, and patients were treated with erythropoiesis- stimulating
agents. The study endpoints are commencement of long-term dialysis and all-cause
mortality for ACRI/ ARB users vs. nonusers. In a median follow-up of 7 months, 20,152 patients (70.7%) required long-term dialysis
and 5,696 (20.0%) died before progression to end-stage renal disease requiring
dialysis. Use of ACEIs/ARBs was associated with a lower risk for long-term dialysis (HR, 0.94 [95% CI, 0.91-0.97]) and the composite out- come of
long-term dialysis or death (0.94 [0.92-0.97]). The renal benefit of ACEI/ARB
use was consistent across most patient subgroups, as was that of
ACEI or ARB monotherapy. Compared with nonusers, the ACEI/ARB users had a
higher hyperkalemia-associated hospitalization rate, but the risk of predialysis mortality caused
by hyper- kalemia was not significantly increased (HR, 1.03 [95% CI,
0.92-1.16]; P = 0.30). We concluded that patients with stable hypertension and
advanced CKD who receive therapy with ACEIs/ARBs exhibit an association with
lower risk for long-term dialysis or death by 6%. This benefit
does not increase the risk of all-cause mortality.
Key Words: aldosterone, angiotensin, chronic kidney disease,
hypertension, renin
S02 The Mechanism Underlying LPS-Induced Adrenal Insufficiency
Changnan Wang, Xiaoyan Zhu, and Xin Ni
Department of Physiology, Second Military Medical University, Shanghai 200433
nixinsmmu@hotmail.com
Reversible adrenal insufficiency, which is characterized by
low basal cortisol levels or low cortisol
responses to adrenocorticotropic hormone (ACTH) stimulation, has
frequently been diagnosed in critically ill patients
with sepsis, and is always associated with
higher mortality rates. However, the pathogenesis of
sepsis-induced adrenal insufficiency remains largely unknown. In
animal model of endoxemia, we showed that
systemic administration of lipopolysaccharide (LPS) causes mitochondrial
oxidative stress, thereby leading to adrenocortical insufficiency.
Since hydrogen sulfide (H2S) and nitric oxide (NO) have been
implicated to be involved in mitochondria damage
in various tissues, we then examined whether
NO and H2S contribute to mitochondrial oxidative stress in
adrenal cortex and adrenocortical insufficiency during endotoxemia.
Systemic administration of LPS increased iNOS expres-
sion and NO production whereas inhibited the
expression of H2S generation enzymes cystathionine-beta-synthase (CBS) and
cystathionine-gamma-lyase (CSE) and H2S production in adrenal glands of mice.
Specific iNOS in- hibitor 1400W significantly attenuated
the LPS-induced mitochondrial superoxide production and dysfunction
in adrenal glands, meanwhile reversed the LPS-induced
adrenocortical hyporesponsiveness to ACTH. In contrast, administration
of NO donor sodium nitroprusside (SNP) led
to mitochondrial oxidative stress and dysfunction in
ad- renal glands, which resulted in blunted
corticosterone response to ACTH. Administration of H2S donor GYY4137 inhibited NO production and
reversed LPS-induced adrenocortical hyporesponsiveness. Either CBS/CSE in-
hibitors or siRNAs led to mitochondrial oxidative
stress and dysfunction, meanwhile resulted in blunted
corticosterone responses to ACTH. Local suppression of
CBS or CSE in adrenal glands significantly
increased the mortality in endotoxemic mice, which
was also improved by GYY4137. Our data
suggest that reduced production of H2S and overproduction of NO contributes to
mitochondrial oxidative stress in adrenocortical cells and
subsequently leads to adrenal insufficiency. Endogenous H2S generation can be a critical regulator of
adrenocortical responsiveness, and might lead to the development of new therapeutic approaches
for the treatment of relative adrenal insufficiency during sepsis.
Key Words: glucocorticoids, adrenal gland, nitric oxide,
hydrogen sulfide
S03 The Mechanism of Pro-Adhesion and Pro-Survival Effects of Glucocorticoid in Human Ovarian Cancer Cells
Lijuan Yin1, Fang Fang2, Yan Wang1, Gaoxiang Huang1, Ning Hui2, and Jian Lu1
1Department of Pathophysiology
2Department of Obstetrics and Gynecology, Changhai Hospital; the Second Military Medical University, Shanghai 200433
lujian326@163.com
Cell adhesion to extracellular matrix (ECM) is
controlled by multiple signaling molecules and intracellular
pathways, and is pivotal for survival and
growth of most of the solid cancer
cells. Our previous works demon- strated that
dexamethasone (DEX), a synthetic glucocorticoid (GC), significantly increased cell adhesion
to ECM by increasing levels of integrin b1, a4 and a5, thereby enhancing cell resistance to chemotherapeutics
in human ovarian cancer cells. However, it
is unclear whether components of ECM or
other membrane molecules besides integrin b1 are also involved in pro-adhesion effect
of DEX in ovarian cancer cells. Therefore,
in the present study we investigated the
effect of DEX on the expression of
the components of ECM, CD44 and mucin
antigen 1 (MUC1), a large transmembrane epithelial mucin glycoprotein by Real-time
PCR, Western blot and ELISA. The results demonstrated that treatment of cells
with 100 nM DEX did not change the expression of collagens
(I, III and IV), laminin, CD44 and its principal ligand hyaluronan (HA), but significantly up-regulated the expression of fibronectin (FN) and
MUC1 in ovarian cancer cells. Inhibiting the expression of FN with specific small RNA interference (siRNA) or treatment
of ovarian cancer cells with CD44 blocking antibody significantly attenuated the
pro-adhesion of DEX, suggesting that up-regulation of FN may mediate the pro-adhesion effect of DEX via interaction with CD44 besides with its primary
receptor integrin a5b1. Moreover, DEX significantly enhanced cell resistance
to cytotoxic drug, paclitaxel (PTX) by activating PI-3K-Akt pathway. Finally, we found
that inhibiting the expression of MUC1 with siRNA significantly attenuated the DEX-induced effects of pro-adhesion, Akt activation and pro-survival. In
conclusion, these results provided new data that up-regulation of FN and
MUC1 by DEX is involved in the DEX-induced effects of pro-adhesion and protecting ovarian cancer cells from chemotherapy.
Key Words: dexamethasone, cell adhesion, fibronectin, mucin
1, Akt, cell survival
S04 Serum Levels of Chemerin and Lp-PLA2 and Low-Grade Inflammation Were Decreased in Obese Female Juveniles by
Short Term Aerobic Exercise Plus Dieting
Xiaohui Wang
Department of Kinesiology, Shanghai University of Sport, Shanghai 200438
wangpan96@126.com
To determine the effects of short-term moderate
intensity aerobic exercise plus dieting on serum
chemerin, lipoprotein-associated phospholipase A2 (Lp-PLA2) and low-grade
inflammation in obese female juveniles. Fifty obese
female juveniles were divided into exercise plus
dieting group (n = 30) who received 4-week aerobic
exercise plus dieting or dieting group (n = 20). Before and after experiments, anthropometric index, metabolisms of glucose and lipid and serum inflammatory indicators
were measured. The serum levels of chemerin, Lp-PLA2, CRP, IL-6, IL-1b, TNFa and leptin declined and adiponectin increased in
exercise plus dieting group, accompanied with reduc-
tion in weight, BMI, waist circumference, body
fat, fasting blood glucose, fasting insulin, HOMA-IR,
total cholesterol (TC), triglycerides (TG) and LDL
and enhancement in HDL. Dieting yielded small
loss of weight, BMI, waist cir- cumference
and body fat while no measurable health
benefits were found. Correlation analysis showed the
cor- relation of chemerin with glucose metabolic
index, TG and Lp-PLA2, and Lp-PLA2 with
inflammatory markers and TG. In conclusion, it
is the first report that short term
aerobic training plus dieting decreased serum chemerin
and Lp-PLA2 and low-grade inflammation in obese
female juveniles, accompanied with significant fat loss
and im- provement of cardio-metabolic risk factors.
Key Words: chemerin, Lp-PLA2, low-grade inflammation, obese female
juveniles, aerobic training plus dieting
S05 Social Buffering Effects on Corticosterone, the Stressor-Produced Decreases in Dendritic Length,
Branches of the Dentate Granule Cells and Related Memory Deficits
Lung Yu
Institute of Behavioral Medicine, College of Medicine, National Cheng Kung University, Tainan 70101
lungyu@mail.ncku.edu.tw
This study was undertaken to assess whether
a robust stressor regimen may render morphological changes in the existing DG granule cells and
whether the presence of companions can reverse the plausible changes. Moreover,
we assessed whether the morphological changes in DG granule cells could
be associated with the altera- tions in the performance of the
DG-related working memory. Single granule cell was visualized by lucifer yellow
labeling. Six hours after the conclusion of the stressor procedure, the
morphological indices of the granule cells along the neuraxis (from Bregma
-0.94 to -3.5) of the DG were obtained in three groups
of Balb/C mice, the stressor-free control, stressor, and stressor with company
groups. The stressor procedure produced significant decreases in the total length
of dendrites, number of dendritic branches, and the size of the
dendritic field in granule cells, while companions reversed all these changes.
Social buffering effect did not affect the stressor- enhanced corticosterone. The
stressor-produced decreases and companions-exerted reversals in the above- mentioned morphological indices
of the DG granule cells were correlated positively with the working
memory per- formance in the object recognition task.
Key Words: granule cells, dendritic field, dentate gyrus
S06 Influence of Berberine on Testosterone Secretion from
Mice Testicular Leydig TM3 Cell
Lu-Ping Tseng, Ju-Wen Yu, Ying-Chi Chen, Yi-Jing Chen, Pin-Yi Liu, and Shyi-Wu Wang* Department of Physiology & Pharmacology, College of Medicine, Chang Gung University, Kweisan Taoyuan, Taiwan
swwang@mail.cgu.edu.tw
Berberine is a traditional Chinese medicine that has a long history and is widely used in many
countries. Research indicates that the pharmacological effects of berberine including antioxidation,
treatment of gastrointestinal discomfort, improve cardiovascular system, anti-inflammatory, anti-bacterial, anti-microbial,
anti-viral and anti-tumor. The main purpose of this study is to investigate the effect of berberine on testosterone secretion from testicular Leydig TM3
cells. Results showed that: (1) testosterone secretion form TM3 cells is in a time-dependant manner;
(2) berberine induced testosterone secretion from TM3 at 24
h; (3) berberine increased StAR, P450scc, P450c17,
and 17b-HSD proteins expression in TM3 cells after
24 h; (4) decreased phosphorylation of p38
and enhanced ERK1/2 weres found in TM3
cells treated with berberine for 5 min. In
conclusion, our result showed that stimulated testosterone secretion from Leydig TM3 cells treated
with berberine might be through the increased steroidogenic enzymes and expression
of phosphorylated ERK1/2 and p38.
Key Words: berberine, Leydig cells, TM3 cells, testosterone,
mice
S07 Pancreatic Dopamine Elicits a Somatostatin-Dependent Inhibition of Insulin Secretion through Activating
Dopamine Receptor 2
Jin-Xia Zhu
Department of Physiology and Pathophysiology, School of Basic Medicinal Sciences Capital Medical University, Beijing 100069
zhu_jx@ccmu.edu.cn
Dopamine regulates insulin secretion. Dopamine receptor 2 (D2 receptors) is able to regulate the level
of blood glucose. But the mechanism is
unclear. We have reported that D2 receptors were constant expressed in the so-
matostatin secreting d cells, but not in the insulin secreting
b cells in rat islets. We hypothesized that dopamine may regulate insulin secretion through regulating somatostatin release from
d cells by binding with D2 receptors. Immunofluorescence, HPLC/MS, radioimmunoassay, pancreatic tissue incubation, and
islets isolation etc. were employed. We demonstrated that 1) dopamine was abundantly existed in the pancreatic tissue, D2 receptors and
somatostatin receptor 2 (SSTR2) were respectively distributed in the d and b cells in rat islets, 2) in vivo study,
activating D2 receptors increased blood glucose, decreased serous insulin and pancreatic
insulin, and increased pancreatic somatostatin, 3) in
the isolated pancreatic tissue and islets, activating
D2 receptors inhibited insulin secretion and promoted
somatostatin secretion. The D2 agonist-induced inhibition of insulin secretion was blocked by SSTR2
antagonist. This study demonstrates that pancreatic dopamine
is able to elicit a somatostatin-dependent inhibition of insulin
secretion through activating D2 receptors.
Key Words: D2 receptors, somatostatin, insulin secretion
S08 Improvement of Myocardial Energy Metabolism Participates in the Cardioprotection of Chronic Intermittent
Hypobaric Hypoxia
Hui-Jie Ma1, Yan Liu2, Yue Cao2, Xu-Yi Li1, Shuang-Qiao Xu1, and Yi Zhang1
1Department of Physiology, Hebei Medical University, Shijiazhuang, 050017
2Department of Endocrinology, Third Hospital of HebeiMedical University, Shijiazhuang 050051, Hebei
lily564300@163.com
Previous studies demonstrated that chronic intermittent hypobaric
hypoxia (CIHH) protects heart against ischemia/reperfusion injury
in rat via different mechanisms. We hypothesized that CIHH may protect the heart via improvement of the energy metabolism of
the heart. The aim of present study
is to investigate the role energy metabolism
in cardioprotection of CIHH. Male Sprague-Dawley rats
received CIHH treatment or no treatment (control)
in a hypobaric chamber simulating 5000-meter altitude for 28 days. Fast
blood glucose, blood lipid and glucose tolerance were measured. The left
ventricular function of isolated hearts was evaluated during 30 min of
ischemia and 60 min of reperfusion. Gene expression involved in cardiac
energy metabolism was determined using quantitative PCR and Western blot. The
results are as follows: 1. There were no differences of fast
blood glucose, blood lipid and glucose tolerance between control and CIHH
rats. 2. The recovery of cardiac function after I/R was improved
in CIHH rats compared with control rats (P < 0.05). 3. CIHH significantly increased cardiac p-AMPK/AMPK ratios, PGC-1a, and GLUT4, but decreased PDK4 gene
expression at both mRNA and protein level
(P < 0.05). We concluded that CIHH could ameliorate ischemia/reperfusion injury by increasing cardiac p-AMPK, PGC-1a, and GLUT4 expression, but reducing PDK4
expression in cardiomyocyte. (Supported by the National
Natural Sciences Foundation of China: No. 30572086,
81200070, 81100607 and the National Basic Research
Development Program of China: No. 2012CB518200).
Key Words: chronic intermittent hypobaric hypoxia, cardioprotection, ischemia/reperfusion,
energy metabolism, rat
S09 Improving Insulin Sensitivity inType II DM Rat via Long Term Oral Administration Lactic Acid Bacteria
Hui-Yu Huang
Department of Food Science, Nutrition, and Nutraceutical Biotechnology, Shih Chien University, Taipei, Taiwan
maggieh323@hotmail.com
The major recent challenge in medical or
food science in developed/developing counties is to
combat against the diet related disorders, especially
diseases connected to insulin resistance (IR) syndrome.
IR represents a cluster of metabolic disorders, including obesity, glucose intolerance and predisposes to type 2 diabetes. Probiotics are live organisms that
are primarily used to improve gastrointestinal disorders such as diarrhea, irritable
bowel syndrome, constipation, lactose intolerance, and to inhibit the excessive proliferation
of pathogenic intestinal bacteria. How- ever, recent studies have suggested that
probiotics could have beneficial effects beyond gastrointestinal health, as they were
found to improve certain metabolic disorders such as hypertension, hypoglycemia, improvement
and/or treatment of lipid profiles, modulation of insulin resistance and sensitivity.
Certain strains of LAB, particularly strains from the genera Lactobacillus showed several health promoting effects, including antidiabetic property. L. plantarum 299v,
L. plantarum K68, L. reuteri strain 263 (Lr 263) and
L. reuteri ADR-1(ADR-1). Our results indicate that the hypoglycermia effect of Lactobacillus in STZ induced type 2 DM is associated with alleviated fatty acid oxidation genes expression and suppressed fatty acid synthesis
genes expression, resulting in enhanced insulin sensitivity. These findings suggest a therapeutic potential of probiotics for prevention and treatment
of type 2 diabetes.
Key Words: insulin sensitivity, type II DM,
lactic acid bacteria
S10 Adipokines: Novel Therapeutic Target of Rheumatoid Arthritis
Chih-Hsin Tang
Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan
chtang@mail.cmu.edu.tw
Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by robust infiltration of leuko- cytes into the synovium,
resulting in hyperplasia of the synovial lining, progressive cartilage destruction, and
ero- sion of the underlying bone. Articular adipose tissue is a ubiquitous
component of human joints, and adipokines (adiponectin, leptin and resistin) are
protein hormone secreted predominantly by differentiated adipocytes and involved in energy
homeostasis. We found adipokines in synovial fluid and tissue from patients with RA were overexpressed. In
addition, adipokines promoted inflammatory cytokine IL-6 production in human RA synovial
fibroblasts (SF). Adipokines also increased matrix metalloproteinases (MMP)-3 in human chondrocytes
may con- tribute to the breakdown of articular cartilage during arthritis.
Furthermore, adipokines directly induced a signifi- cant increase in expression of vascular endothelial growth factor (VEGF) in endothelial progenitor cells
(EPCs) and promoted EPCs homing into the synovium during RA angiogenesis.
These findings provide support for adi- pokines as a therapeutic target for the patients with RA.
Key Words: adipokines, rheumatoid arthritis, adipocytes, IL-6, matrix metalloproteinases
(MMP), vascular endothelial growth factor (VEGF), endothelial progenitor cells (EPCs)
S11 An Investigation of Endocrine Abnormalities’ Influence on the Psychological, Neurological, and Immunological Health of
People with Metabolic Syndrome
Amy H.T. Davis1, Ching-Ling Lin2, Paulus S. Wang1, and Yu-Ju Chen2
1China Medical University Hospital, Taichung 2National Defense Medical Center, Taipei, Taiwan judychen@mail.ndmctsgh.edu.tw
Endocrine abnormalities have profound and widespread effects
on a person’s overall health status. Metabolic syndrome is a recognized clinical cluster of endocrine abnormalities, and it is a prevalent health problem around
the world. Over the last decade, despite advances in diagnostics, new pharmaceuticals, and invasive surgical interventions to treat this cluster of
endocrine abnormalities; metabolic syndrome is more prevalent than even be- fore.
The primary consequence of metabolic syndrome – Cardiovascular Disease (CVD) is now the leading cause of death for men
and women. Clearly, more work is needed in understanding metabolic syndrome to discover effective intervention
targets. Given the widespread effects of endocrine abnormalities on a person’s
overall health, the objective of this study is to investigate the influence
of endocrine abnormalities on the psychological, neurological, and immunological health of
people with metabolic syndrome. A convenience sample from two clinics in northern Taiwan was recruited for this study. Psychological health included perceived susceptibility of CVD, per- ceived stress, personality, and depression. Neurological health included measures of sympathetic, parasympathetic, and an
interplay between these two nervous systems. Immunological health included pro-inflammatory markers
of CRP, IL6, and TNF-a.
Of the 405 subjects recruited, 345 had
complete data for analyses (85.2%).
Metabolic Syndrome
|
Variables
|
Yes, n = 94 (27.2%)
|
No, n = 251 (72.8%)
|
|
Age
Psychological health
Perceived Susceptibility of CVD**
|
37.4 ± 10.8
10.3 ± 2.9
|
34.0 ± 12.8
8.4 ± 3.3
|
|
Perceived stress Conscientiousness personality* Depression**
Neurological health Sympathetic- LF nu* Parasympathetic- HF nu*
Sympathetic + Parasympathetic-lnTP**
|
23.1 ± 6.7
19.7 ± 3.6
11.7 ± 9.1
43.3 ± 19.0
6.2 ± 1.0
|
22.6 ± 6.6
20.6 ± 3.7
8.6 ± 7.1
48.1 ± 20.0
6.6 ± 0.9
|
|
Immunological health C- Reactive Protein** IL-6
TNF-a
|
1.28 ± 1.19
3.09 ± 8.16
0.06 ± 0.15
|
0.70 ± 0.94
1.75 ± 4.95
0.06 ± 0.18
|
*P < 0.05, **P < 0.01
Our findings elucidated the widespread influence of
endocrine abnormalities on the psychological, neurologic, and
immunological of people with metabolic syndrome, thus,
providing new intervention targets for early prevention
of cardiovascular disease.
Key Words: metabolic syndrome, psychological health, neurological health, immunological health, cardio- vascular disease, endocrine
S12 Roles and Mechanisms of the Novel Estrogen Receptor GPR30 on MDA-MB-231 Breast Cancer Cells Migration
and Invasion Induced by Estrogen
Kewen Zhou, Chen Wang, Ping Li, Xiaoyu Wang, Zhiyuan Long, Ling Wang, and Ting-huai Wang Department of Physiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou wangth@mail.sysu.edu.cn
ER-negative breast cancer is an aggressive breast
cancer subtype characterized by poor clinical outcome,
and tumor spread and distance metastasis is
the major cause of death of these
patients. Researchers suggest that a major of tissues or cells
of ER-negative breast cancer are GPR30 positive on different levels. Here,
we aimed to discuss the roles and mechanisms of the novel estrogen receptor GPR30 on MDA-MB-231 breast cancer cells migration
and invasion induced by estrogen. MDA-MB-231cells is triple-negative breast cancer cell
that belong to one subtype of ER-negative breast cancer. In MDA-MB-231 cells, wound healing assays, Transwell assays and western
blot were used to detect the cell migration, invasion and phosphorylated AKT levels, respectively. When blockade GPR30 signaling cannot abrogate the migration and
invasion of estrogen in MDA-MB-231 cells, pre- incubated with an ERb antibody peptide or stimulated ERb signaling with an agonist, DPN can attenuate
or mimic the migration and invasion of
estrogen, respectively. In addition, co-incubated DPN with
the GPR30 agonists, G1 or 4-OHT both down-regulated ERβ protein and phosphorylated AKT levels, indicating an antagonist effect be- tween GPR30 and ERβ signaling in MDA-MB-231 cells. Our study provides the evidence that estrogen involves in migration and invasion of MDA-MB-231 breast
cancer cells. Moreover, estrogen through GPR30 may
exert distinct effects dependent on the ratio
between GPR30 and ERb in target ER-negative cells.
Key Words: estrogen, ER-negative breast cancer, GPR30,
ERb, cell motility
S13 Effects of Caloric Restriction on Liver Function and
Oxidative Stress in Male Rats
Mei-Mei Kau1, Sheng-Ju Chuang2, Kai-Lee Wang3, Chih-Chieh Chen4, Yi-Ting Hsieh4, and Paulus S. Wang4, 5
1Department of Nursing, National Taipei University of Nursing and Health Sciences, Taipei
2Department of Sports Sciences, University of Taipei, Taipei
3National Environmental Health Research Center, National Health Research Institutes, Taipei
4Department of Physiology, National Yang-Ming University, Taipei
5Medical Center of Aging Research, China Medical University Hospital, Taichung, Taiwan
meimei@ntunhs.edu.tw
Caloric restriction (CR) can expand lifespan and
improve many health status including obesity, cardiovas-
cular disease, type II diabetes and cancers.
However, the benefits and harms of different
levels of CR are still unclear. The
purpose of this study was to determine
the effects of strict CR on liver
function and oxidative stress in male rats.
Twenty-four male rats were placed into metabolic
cage and divided into three groups: control
group (standard rat chow diet al libitum), CR 50% group and CR
75% group (50% and 75% reduction of
the food in- take al libitum) for 6 days. CR (50% or 75%)
for 6 days produced obviously reduction in body weight, blood glu- cose,
plasma triglyceride (TG) and liver weight but did not alter the
levels of aspartate aminotransferase (AST or SGOT) and alanine aminotransferase (ALT or
SGPT). There were no significant differences in lipid peroxidation of plasma
and liver among ad libitum and CR groups. However, levels of
protein carbonyl in liver were increased and total antioxidant capacity was decreased in CR groups. We also found that
CR decreased water intake and urine output and elevated the levels of plasma corticosterone
and aldosterone. In summary, our study demon- strated that CR for 6
days resulted in beneficial effects on health, e.g. weight loss, hypoglycemic effect and re- duced TG.
Elevated plasma aldosterone and corticosterone may play an adaptive role in
energy deprival state and may contribute to the balance of electrolytes and blood glucose. Although liver weight reduced,
SGOT and SGPT remained unchanged in CR groups. We speculate that the reduction of liver weight may be associated with glycogen depletion,
apoptosis or autophagy rather than cell necrosis. CR increased hepatic protein
oxida- tive stress and decreased total antioxidant capacity, however, did not
alter the lipid peroxidation as compared with ad libitum group. Both
the hepatic apoptosis and/or autophagy is/are induced by CR and the hepatic oxida- tive stress
is involved in this process remain to be established.
Key Words: caloric restriction, oxidative stress, liver
function, corticosterone, aldosterone, weight loss
S14 Inhibitory Effects of Roscovitine on Androgen Receptor and Proliferation of Prostate Cancer Cells
Fu-Ning Hsu1, Mei-Chih Chen2, and Ho Lin3
1Department of Molecular and Cellular Medicine, Texas A&M University, Texas 77843, U.S.A. 2Department of Medical Research, Taichung Veterans General Hospital, Taichung 40705 3Department of Life Sciences, National Chung Hsing University, Taichung 40227, Taiwan hlin@dragon.nchu.edu.tw
It has been reported that cyclin-dependent kinases
play important roles in modulating androgen receptor
(AR) function and proliferation of prostate cancer.
Roscovitine is a specific Cdk inhibitor and had been applied in clinical trials and combination therapy
in many types of cancer. The aim of this study is
to investigate whether AR is a target of Roscovitine and therefore affects prostate cancer cell growth. Roscovitine was treated in
culture medium of prostate cancer cell lines (LNCaP, LNCaPdcc, 22Rv1) as well as normal prostatic epithelial cells. Cell growth (in vitro and in vivo) and AR activation (including localization, reporter assay,
PSA (prostate-specific anti- gen) expression) were evaluated. Roscovitine treatment resulted in
significant growth inhibitions in both prostate cancer cell lines and normal
prostatic cells. The results of xenografted tumor growth also support the
finding. The indices of AR activation, including subcellular localization, promoter activity,
and PSA production/secretion, were all inhibited after Roscovitine treatment. In conclusion,
AR might be one of the pharmaceutical targets of Roscovitine in prostate cancer cells and, therefore, Roscovitine might be a potential
drug candidate in prostate cancer therapy.
Key Words: androgen receptor, cell proliferation, prostate
cancer, roscovitine
S15 Estrogen Suppresses Age-Related Microglia Activation and Dopaminergic Neuron Loss in the Substantia Nigra
Yu-Min Kuo
Department of Cell Biology and Anatomy, National Cheng Kung University, Tainan, Taiwan
kuoym@mail.ncku.edu.tw
Age-related dopaminergic (DA) neuron degeneration in the
substantia nigra (SN) is the pathological hallmark
of Parkinson’s disease. The incidence of Parkinson’s
disease is higher in men than in
women and DA neurons are highly susceptible
to microglia activation and pro-inflammatory cytotoxic factors.
This study aimed to investi- gate the
effect of sex on age and inflammation-induced
microglia activation and DA neuron death. The
numbers of tyrosine hydroxylase-positive DA neurons and
degrees of microglia activation in the SN
were determined in different ages (3, 6,
9 and 12 months) of male and female mice. The degrees
of LPS-induced microglia activa- tion and DA neuron loss in the
SN were compared between male and female mice. Bilateral ovariectomy was
employed to study the roles of ovarian hormones in microglia activation and DA neuron loss. The effects of 17β- estradiol on levels of microglia activation-associated markers
and activities of inwardly rectifying K+ channel were
further investigated in a mouse microglia cell (BV2) line. The degrees of microglia activation and DA neuron loss
were increased with age in both sexes and were more pronounced
in male than in female mice. Likewise, peripheral LPS-induced microglia activation
and DA neuron loss were also more prominent in male mice
than in females. Ovariectomy for three months (from 6- to 9-month-old)
enhanced microglia activation and DA neuron loss. Supplement of 17b-estradiol immediately after ovariectomy for three months
almost completely antagonized the ovariectomy effect, while
supplements of 17b-estradiol two months after ovariectomy for one
month (from 8- to 9-month-old) partially reversed
the degrees of microglia activation, but not
the lost DA neurons. Pretreatment of BV2
microglia cells with 17b-estradiol suppressed the LPS-induced elevations of Toll-like
receptor 4, phosphorylation
of p38 and TNF-a levels. The activities of microglia inwardly rectifying K+ channel were inhibited by 17b-estradiol.
Age-related microglia activation and DA neuron loss
were more dramatic in the male mice
than the female mice. The sex difference
in DA neuron loss is partially derived from the anti-microglia
activation of ovarian estrogen.
Key Words: estrogen, age-related, dopaminergic neuron, microglia,
substantia nigra
S16 Protective Effect of Chronic Intermittent Hypobaric Hypoxia on
Metabolic Syndrome Rats
Jing-Jing Zhou, Fang Yuan, Hui-Jie Ma, and Yi Zhang
Department of Physiology, Hebei Medical University, Shijiazhuang 050017, Hebei
zhyhenryphy@163.com, zhyhenry@hotmail.com
Accumulating evidence demonstrate that chronic intermittent hypobaric
hypoxia (CIHH) has a variety of ben- eficial effects
on the body, such as cardiac protection, brain protection, anti-hypertension, and immune
regulation. The aim of this study was to test the hypothesis
that CIHH could have protective effect on metabolic syndrome (MS). Adult
male Sprague-Dawley rats were used and metabolic syndrome (MS) rats were
induced by 10% fruc- tose in drinking water for 6 weeks in
this study. The rats were treated with hypobaric hypoxia simulating 5,000 m altitude in a hypobaric chamber, 6 h per day for 28 days. Four parts of
experiments, including cardioprotec- tion, antiarrhythmia, hepar protection and skeletal protection of
CIHH, were performed in the study. The results showed: 1. CIHH
treatment protected the heart against ischemia/reperfusion-induced injury, promoted the recovery of cardiac function from ischemia/reperfusion, diminished the mitochondria-related apoptosis of myocardium, and
prevented cardiac hypertrophy, elevation of arterial pressure, and abnormal serum levels
of glucose, fasting insulin, insulin C peptide, triglyceride and cholesterol in
MS rats; 2. CIHH treatment protected the heart against arrhythmia induced by
ischemia/reperfusion and prevented the abnormal change of action potential during ischemia/reperfusion
in MS rats; 3. CIHH treatment protected the liver against nonalcoholic
fatty liver disease (NFLD) by preventing the up-regulation of endoplasmic reticulum
stress (ERS) proteins including GRP78 (glucose-regulated protein78), CHOP (C/EBP Homologous Protein), and caspase-12 in hepatic tissue of MS rats; 4. CIHH
treatment protected the skeletal muscle by inhibiting inflammation and preventing the
up-regulation of TNFa (tumor necrosis factor a) and MCP-1 (monocyte chemoattractant protein-1) as
well as NF-kappa B (nuclear factor kappa beta) in skeletal muscle of MS rats. It suggests that
CIHH could protect heart, liver and skeletal muscle in MS rats.
Key Words: chronic intermittent hypobaric hypoxia, metabolic
syndrome, cardioprotection, antiarrhythmia, NFLD, ERS, insulin resistance,
TNFa
S17 Protective Effects of Curcumin on Hepatic Fibrosis
Mu-En Wang and Chih-Hsien Chiu
Laboratory of Animal Physiology, Department of Animal Science and Technology,
National Taiwan University, Taipei, Taiwan
chiuchihhsien@ntu.edu.tw
Hepatic fibrosis describes a condition of excessive collagen accumulated in the liver tissue upon hepatic in- jury. It could
be caused by various factors such as infections, alcohol and toxic
chemical intake. Under these stimuli, damaged hepatocytes initiate the inflammatory response
and hepatic stellate cell (HSC) activation to synthesize and secrete collagen.
Previous studies have indicated the protective effects of curcumin on many
diseases and cancers via its anti-oxidant, anti-inflammatory and apoptosis-promoting effects. Our laboratory specifically studies the molecular mechanisms
of curcumin mediated anti-hepatic fibrosis effects. By using in vivo and
in vitro study models, we have shown that curcumin could attenuate
hepatic fibrosis via promoting damaged hepatocytes un- dergo apoptosis rather than necrosis. In male BALB/c mice,
we found that curcumin decreased TAA induced liver inflammation, HSC activation,
but increased apoptotic cell death. Moreover, we found that curcumin upregulated
P53 protein expression level, and down-regulated Bcl-2 mRNA level in vivo.
Similarly, the apoptosis enhancing ef- fects of curcumin was also found in AML12 mouse hepatocytes. Taken together, our preliminary data suggest that curcumin might eliminate liver
injury induced hepatic fibrosis via shifting damaged hepatocytes from necrosis to apoptosis, and thus reducing the inflammatory response and HSC activation. Our team is
currently studying how curcumin regulates damaged hepatocyte undergo apoptosis. Especially, we
are focusing on the role of EP300, which has been shown
to be inhibited by curcumin, and involved in regulation of P53
activity and stability. Our study extends the understanding of hepatic fibrosis, also provides novel molecular target for clinical therapy.
Key Words: apoptosis, curcumin, hepatic fibrosis, TAA,
EP300
S18 Proteomics Analysis of Hepatic Steatosis in Ovariectomized Rats as a Model of Postmenopausal Status
Chen-Chung Liao1, Yu-Tang Tung2, and Chi-Chang Huang2
1Proteomics Research Center, National Yang-Ming University, Taipei 11221, Taiwan
2Graduate Institute of Sports Science, National Taiwan Sport University, Taoyuan 33301, Taiwan
john5523@ntsu.edu.tw
Postmenopausal women are particularly inclined to an
increased risk of developing non-alcoholic fatty liver
disease (NAFLD). The goal of the current
research was to determine the impact of
postmenopausal-induced NAFLD (PM-NAFLD) in an ovariectomized rat
model. Sixteen six-week-old Sprague-Dawley female rats were
randomly separated into two groups (8 per
group), the rats were either sham-operated (Sham)
or bilaterally ovariectomized (Ovx). Four months after
surgery, indices of liver damage and liver
histomorphometry were measured. Both serum AST and
ALT levels were significantly higher in the
Ovx than those in the Sham control.
In parallel, we carried out quantitative LC-MS/MS-based
proteomic profiling of livers from rats with PM-NAFLD: 1) to provide
baseline knowledge of the PM-NAFLD proteome and 2) to use bioinformatics
tool, Ingenuity Pathways Analysis (IPA), in addressing the cellular and metabolic
processes or affecting certain metabolism of drugs and molecular functions in
which proteins involved in PM-NAFLD. Of the 586 identified proteins, 59
(10.0%) and 48 (8.2%) protein expres- sions were significantly higher and
lower, respectively, in the Ovx group than in the Sham group.
Data inter- pretation is accomplished by comparing the results between Sham
and Ovx rats. In conclusion, the changes in regulation of proteins
implicated in PM-NAFLD may affect other vital biological processes in the
body apart from causing postmenopause mediated liver dysfunction. Our quantitative proteomics
analysis may also provide po- tential biomarkers and further clinical applications
for PM-NAFLD.
Key Words: ingenuity pathways analysis, liver, proteome
S19 Is It Possible to Generate a Bio-Efficient Human Growth Hormone Variant by Increasing Ligand-Receptor Affinity?
Fangyu Zhao, Yu Zhang, Zhao Zhao, Yan Shen, Xiaoya Xu, Yang Wang, Jian Song, and Yu Wan School of Basic Medical Sciences and Center for Medical Research, Wuhan University, Wuhan 430071, Hubei wanyu@whu.edu.cn
Whethe the bio-efficiency of human growth hormone
(hGH) can be enhanced by increasing the
hormone- receptor affinity is in debating. Previous
studies by Genentech group showed that a
400-fold increase in affin- ity of hGH for its receptor (hGHR) at site 1,
or a 40-fold increase at site 2 was fail to enhance the
biopotency of the hormone; they suggested that the affinity of hGH-hGHR, at least at site 1,
is overmaximized for biopotency. In the present study, using site-directed mutagenesis, we generated mutations of hGH at site1 (R167K, L45D, S184F),
site2 (I4V, R16L, N109Y and D116F) and both (R167K/S184F, R167K N109Y, L45D/N109Y, L45D-R64D),
respectively. The recombinant hGH mutants were prepared by
genetic engineering techniques. The affinity and kinetics
of hGH-WT or mutants binding with hGH
binding protein (hGHBP) were detected with radio-immunoprecip-
itation assays (RIA) and surface plasmon resonance
analysis (SPR). The biopotency of the analogues
was esti- mated both in vitro and in vivo with cell proliferation assays and hGH replacement
therapy in pituitary GH-deficient rats. It was
found that an enhanced Site1 affinity (R167K)
by mutagenesis at site1 alone led to
a reduced binding ability at Site2. Similar phenomenon was observed when Site2 affinity was enhanced by single mutagenesis at Site2
(N109Y). Both types of mutants were unable to improve the hormone’s biopotency. Those mutations with a reduced affinity at either Site1 (L45D, S184F) or site2 (I4V, R16L and D116F) showed a decreased biological efficiency. However, the combined mutation with enhancement of both Site1 and Site2 affinities exhibited a
improvement of biopotency both in vitro and in vivo. These results suggest that the affinity of
hGH for hGHR at either site 1 or Sits2 is not overmaximized for biopotency. However only an affinity increase at both sites in
combination (rather than at Site1 or Site2 alone) can improve hGH’s
biopotency.
Key Words: affinity, biopotency, human growth hormone,
hGH-variant, hGHR
S20 Acromegaly and Cardiovascular Diseases
Harn-Shen Chen
Division of Endocrinology and Metabolism, Taipei Veterans General Hospital, Taiwan Faculty of Medicine, National Yang-Ming University School of Medicine, Taipei, Taiwan chenhs@vghtpe.gov.tw
Acromegaly is a chronic debilitating disorder caused by a GH-producing pituitary adenoma. Active acromegaly is associated with a 2- to 4-fold increased
mortality risk, mainly from cardiovascular and malignant disease. Mortality risk has
been related to various GH excess parameters, including serum GH and
IGF-1 concentrations, and to comorbidity at the time of diagnosis. Generally, mortality is still elevated in cohorts
of patients with acromegaly undergoing modern treatment regimens. Cardiovascular disease is
a major contributor to the increased mortality of acromegalic patients. Prolonged growth hormone (GH) excess
can induce myocardial changes. In fact, growth hormone and insulin-like growth
factor-I excess induces a specific cardiomyopathy. The heart is involved from
the very early stages of the disease in which the hyperkinetic
syndrome (high heart rate and increased systolic output) takes place. Frequently,
if the disease is untreated for many years or unsuccessfully treated,
concentric biventricular hypertrophy and diastolic dysfunction can develop and, at least,
lead to diastolic congestive heart failure. These changes include left ventricular
(LV) hypertrophy, diastolic dysfunction, systolic dysfunction during exercise, arrhythmias and heart failure. The
incidence and severity of these cardiac changes are related to dis-
ease activity and disease duration. LV hypertrophy appears to be an early consequence of GH excess. Trans- phenodial adenectomy
(TSA) is the first choice therapy, but about 50% still required
long-term medical treatment. Long-acting somatostatin anologues are currently indicated for the
treatment of acromegaly. Both random GH and IGF-1 are routinely used
to monitor disease activity, and a normal age and sex related serum
IGF-1 and a fasting GH lower than 2.0~2.5 ng/ml are currently accepted
criteria to define biochemical control in acromegaly.
Key Words: acromegaly, growth hormone, insulin-like growth
factor-1, cardiovascular diseases
S21 Inflammation and Increased IDO in Hippocampus Contribute to Depression-Like Behavior Induced by Estrogen Deficiency
Jianqiang Lu
School of Kinesiology, Shanghai University of Sport, Shanghai 200438
lujianqiang@sus.edu.cn
Estrogen deficiency is involved in the development of depression. However, the mechanism underlying estrogen modulates depression-like behavior remains largely unknown. Inflammation and indoleamine-2,3-dioxygenase (IDO)
have been shown to play pivotal roles in various depression models.
The objective of the present study was to investigate whether estrogen
deficiency-induced depression-like behavior is associated with inflammation and IDO activation in
brain. The results showed that ovariectomy resulted in depression-like behavior in
female rats and caused a decrease in 5-HT content and an increase in levels of IDO, IFN-g, IL-6, toll like receptor (TLR)-4 and
phosphorylated NF-kB (p65 subunit) in hippocampus but not
in prefrontal cortex (PFC). 17b-Estradiol (E2) treatment ameliorated depression-like behavior and restored
above neurochemical alternations in hippocampus in ovariectomized
rats. Partial correlation analysis showed that the
levels of phosphorylated p65, IFN-g and IL-6 in hippocampus correlated to serum
E2 level. Our study suggests that estrogen inhibits inflammation
and activates of IDO and maintains 5-HT
level in hippocampus, thereby ameliorating depression-like behavior.
Key Words: depression, estrogen, indoleamine-2,3-dioxygenase (IDO),
inflammation
S22 Cardiac Protection of Chronic Intermittent Hypobaric Hypoxia against Ischemia/Reperfusion Injury
Yi Zhang
Department of Physiology, Hebei Medical University, Shijiazhuang 050017, Hebei
zhyhenryphy@163.com, zhyhenry@hotmail.com
Myocardial ischemia and reperfusion (I/R) is a common
problem in clinic and there is no satisfactory method for prevention or treatment of I/R injury so far. Chronic
intermittent hypobaric hypoxia (CIHH) similar to the con- cept of ischemia
preconditioning IPC or altitude hypoxia adaptation (AHA) has been recognized to
confer a protec- tive effect on heart against I/R injury with a
longer protective effect than IPC and a less adverse effect than AHA. Our research work demonstrated that CIHH increases myocardial tolerance to ischemia
or hypoxia, reserving cardiac function and preventing arrhthmia during I/R. Multiple
mechanisms or pathway underlying the cardiac protection of CIHH have been
proposed, such as increase of electrical stability of cell membrane, induction
of heat-shock protein, enhancement of myocardial antioxidation capacity increase of coronary flow and myocardial capillary angiogenesis, activation of adenosine
triphosphate (ATP)-sensitive potassium channels, inhibition of mi-
tochondrial permeability transition pores, and activation of
PKC and iNOS. In addition, CIHH treatment
is simple, safe, economical and easy to
apply. Thus it has important clinical value.
Key Words: chronic intermittent hypobaric hypoxia, cardiac
protection, anti-arrhythmia, ischemia/reperfusion
S23 Chronic Vagus Nerve Stimulation Ameliorates Endothelial Dysfunction and Inflammation Reaction in
Ovariectomized Rats
Ping Li, Peng Sun, Chen Wang, Xiaoyu Wang, Zhiyuan Long, Ling Wang, and Tinghuai Wang Department of Physiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou wangth@mail.sysu.edu.cn
Cardiovascular diseases (CVDs) are the leading causes
of morbidity and mortality in women, while
the vast majority of CVDs occur in
postmenopausal women, which have been known that
it is closely linked to the endothelial
dysfunction and inflammation reaction. Vagus nerve stimulation (VNS), as a method for activating cholinergic anti- inflammatory pathways, could suppress
endothelial activation and minimizes tissue injury during inflammation. The aim of
this study was to investigate the effects and underlying mechanisms of
chronic VNS on endothelial dysfunc- tion and inflammation responses in ovariectomized
(OVX) rats. Sprague-Dawley rats (7 to 8 months old) were ran- domly
assigned into four groups as follows: Sham-OVX, OVX, OVX+Sham-VNS, and OVX+VNS groups. During the entire experimental period, the OVX+VNS group
received chronic VNS for 3 h (20.0 Hz, 1.0 mA, and 10.00
ms pulse width) at the same time every other day. After 12
weeks of VNS, morphological changes were investigated with light microscope, scanning and transmission electron microscope.
The endothelial function was evaluated by measuring relaxation responses to acetylcholine
(ACh) and the serum Endothelin-1 (ET-1), Nitric Oxide (NO) levels. Molecular
mechanisms were also analyzed with immunohistochemistry, immunofluorescence and enzyme-linked aptamer-sorbent assay
(ELASA). Electron microscopy and light microscopy analyses showed that chronic VNS
prevent the OVX rats from endothelial swollen, desquamate and even necrotic.
Meanwhile, chronic VNS also improved endothelial function in OVX rats by
restoring the endothelial nitric oxide synthase (eNOS) and serum ET-1 levels.
Increased cell adhesion molecule (VCAM-1, ICAM-1 and E-selectin) expression of thoracic
aortas and circulation cytokines (TNF-a, IL-6, MCP-1, and CINC/KC) were encountered
in OVX rats, as well as exces-
sive activation of NF-kB pathway. Nevertheless, chronic VNS significantly enhanced
the content of ACh in the thoracic
aortas and prevented the nuclear translocation of
NF-kB p65. This in vivo study determined
that the administration of chronic VNS, in
the early stage of estrogen deficiency, protected
OVX rats from endothelial impair- ment both
in endothelial morphology and function, also reduced
inflammation responses by inhibition the nuclear translocation
of NF-kB p65. Our present study may indicate
that activating the vagus never could be
a promising and powerful supplement in reducing the risks of suffering from further CVDs
in postmenopausal women.
Key Words: chronic vagus nerve stimulation, cholinergic anti-inflammatory
pathway, endothelial dysfunction, inflammation, ovariectomy
S24 Effect of Over-Expression of DS2 Prolactin Receptor on Transcriptome of Breast Cancer Cells
Dunyong Tan, Jie Zhang, Chao Yu, Xie Nie, and Huijuan Zhang Jishou University Research Center of Medical Sciences, Jishou, Hunan 416000 naturet2002@yahoo.com
DS2 PRLR is a variant of PRLR, in which, the
S2 subdomain (100 aa) of the extracellular domain is missing. In the present study, cultured human breast cancer cells (MCF-7) were transfected with adenovirus carrying LF-PRLR (LF-PRLR cells) or DS2 PRLR (DS2 PRLR cells) cDNA or not
carrying any external DNA (Con). After a futher 48 h- incubation of the cells, total RNAs were extracted by using TRIzol reagent and
followed by RNA-sequencing (RNA-Seq). The RNA-Seq data were up-loaded onto genomebrowser
setup by University of California Santa Cruz (genome.ucsc.edu) and a custome
specific channel was used. The alignment and analysis shows: (1) the genes significantly affected by DS2 PRLR over-expression
are those located at chromesome 11 12 14 16 17 19 20
but not at 4 13 18 21. (2) the genes regulated by
DS2 PRLR includes not only encoding genes, but also non-coding ones including microRNAs and small nucleolar RNAs, etc. 19648 (including 17594 encoding
genes and 2054 non-coding genes), 20247 (including 18093 encoding genes 2154
non-coding genes) and 19500 genes (including 17472 encoding genes and non-coding
2028 genes) were detected in Con, LF-PRLR and DS2 PRLR cells
re- spectively in the present RNA-Seq. (3) Of the whole transcriptome,
192 genes were only detected in DS2 PRLR cells but not
in LF-PRLR and Con cells; however, other 483 genes were expressed
in LF-PRLR Con cells, but not in DS2 PRLR cells; and
other 21287 genes were detected in all the three groups of cells.
(4) several tumour-as- sociated genes (27 genes) were observed to be
expressed in DS2 PRLR cells,but not in LF-PRLR and Con cells.
(5) nine apoptosis-associated genes were detected only in DS2 PRLR cells, but
not in LF-PRLR and Con cells.
(6) MicroRNA were also the regulated target
of DS2 PRLR. Expression of eight microRNA
genes were detected exclusively in DS2 PRLR
cells, but not in other two groups
of cells. (7) DS2 PRLR plays roles
in the regulation of small nucleolar RNA
(snoRNA) expression. Ten snoRNAs were detected
only in DS2 PRLR cells, but not
in the other two groups of cells.
Taking together, we concluded that (1) genes affected by DS2 PRLR includs not only
the encoding but also the noncoding ones; (2) the effect of
DS2 PRLR on transcriptome of breast cancer cells are different from
that of its intact counterpart, LF-PRLR; (3) of the genes affected
by DS2 PRLR, tumour-associated genes, apoptosis-associated genes, microRNA and snoRNA are all included.
Key Words: prolactin, PRLR, DS2 LF-PRLR, breast cancer,
RNA-Seq
P01
Analysis of Cyp11a1 Expression and Function in Olfactory Bulb
Szu-Ying Liu1, 2 and Meng-Chun Hu1
1Graduate Institute of Physiology, National Taiwan University College of Medicine, and 2Department of Respiratory Therapy, Tamsui Mackay Memorial Hospital, Taipei, Taiwan
mengchun@ntu.edu.tw
CYP11A1 encodes the first enzyme of steroidogenesis, cytochrome P450scc. CYP11A1 is expressed in the brain to control the synthesis of neurosteroids that are involved in many neural functions. Because the amount of endogenous mRNA is very low, the expression of CYP11A1 gene in the brain has not been
well characterized. We recently used the 4.4-kb promoter of human CYP11A1 gene to drive Cre recombinase expression in the tissues
that produce steroids. The expression of Cre gene was identifiedin several
brain areas including diencephalon, midbrain, hippocampus and amygdala. In this study,
we further detected the Cre expression in the olfactory system including olfactory and epithelium olfactory bulb. In theolfactory bulb, the Cre expression is specifically only detected in the granule
cell layer. Immunofluoresence analysis showed that Cre-expressed cells are NeuN-positive (neuron
marker) and GFAP-negative (astrocyte marker), indicating these cells are differentiated neurons.
Granule cells release g-aminobutyric acid (GABA) which is synthesized by glutamic acid decarboxylase 65 (GAD65) or 67 (GAD67). Our data showed that most of
Cre-expressed cells co-localized with these two enzymes. Some granule cells express
calcium binding protein calretinin or calbindin. However, these two markers were
not co-localized with Creexpres- sion. We used Cyp11a1 knock out mice
(KO) to explore the function of Cyp11a1 inolfactory bulb. Western blot analysis showed that the protein levels of GAD
65, but not GAD67 are significantly decreased in KO mice. In
con- trast, the protein levels of calretinin, but not calbindin are
significantly increased in KO mice. Olfactory marker protein in glomerulus is
also significantly increased in KO mice. In addition, TUNEL assays showed
that the apoptotic cells in the granule cell layer were significantly
decreased in KO mice compared to wild-type mice. These results suggested
that Cyp11a1 may play a critical role in olfactory system.
Key Words: CYP11A1, neurosteroid, olfactory bulb, granule
cell
P02 Adipocyte Hyperplasia in Endothelin-1-Infused Rats
Chih-Chan Lien1, Jia-Ling Jiang1, Deng-Yuan Jian1, Ching-Fai Kwok2, Low-Tone Ho1, 2, 3, and Chi-Chang Juan1, 3, 4
1Institutes of Physiology, School of Medicine, National Yang-Ming University
2Division of Endocrinology and Metabolism, Department of Medicine and 3Department of Medical Research, Taipei Veterans General Hospital 4Department of Education and Research, Taipei City Hospital, Taipei, Taiwan ccjuan@ym.edu.tw
Endothelin-1 (ET-1), an endothelium-derived vasoconstrictor, have been
demonstrated to inhibit adipocyte differentiation in vitro. However, the effect of ET-1
on adipogenesis in vivo and the mechanism underlying this
activity remain unclear. The aim of the
present study was to investigate the regulatory
mechanism of ET-1 on adipogenesis in vitro and in vivo. 3T3-L1 preadipocytes were used to
explore the mechanisms mediating ET-1 actions on
preadipocyte proliferation and adipocyte differentiation. To
investigate the in vivo effect of ET-1, male Sprague-Dawley
rats were infused with ET-1 or saline
for four weeks via intraperitoneally implanted osmotic pumps and the fat pad weight and adipocyte
size of adipose tissues were measured. The results showed that ET-1 stimulated preadipocyte proliferation and increased the cell number at the mitotic
clonal expansion stage of adipocyte differen- tiation via the endothelin A
receptor (ETAR) and activation of the PKC pathway. ET-1, via ETAR, inhibited adipocyte differentiation partially through an ERK-dependent pathway. Furthermore, no significant difference in the body weight and fat pad weight
were observed in either ET-1- or saline-infused rats. Compared with saline-infused
rats, the adipocyte cell number was significantly increased but the adipocyte
size was significantly decreased in ET-1- infused rats. In conclusion, chronic
ET-1 infusion induced adipocyte hyperplasia and decreased adipocyte size in
vivo, which were associated with ET-1-stimulated preadipocyte proliferation and probably adipocyte
lipolysis, but not ET-1-suppressed adipocyte differentiation.
Key Words: adipocyte, adipocyte differentiation, endothelin-1, obesity
P03 The Investigation of the Elder Healthy Lifestyle Factors
Wei-Yueh Chang
Providence University Physical Education Office, Taichung, Taiwan
wychang@pu.edu.tw
Since August 1993, Taiwan has been being an aging society. In
2008, the popularity of the age at 65 or above accounts
for the 10.03% of all popularity of Taiwan. According to the
statistics in 2005 of Ministry of the Interior revealed, 13% of
elders aged 65 or above encountered the problem of dealing personal
activities, 65% of elders had chronical diseases. 20% of elders had
once stayed in hospital and accepted healed during the past year.
For 32.5% of them, the most worried issue is healthy problem.
Studies pointed out that exercise, good diet habit, and life habits
help the elders achieve a holistic healthy life in terms of physical,
social, and spiritual tran- quility. This study aimed to investigate the elders’ exercise pattern, habit and the insufficient factors for achiev-
ing the holistic health. The results were provided to subjects as
the reference for improving healthy issue. The subjects were from the
students attended the Active Aging Learning programs in Providence university. All
the subjects were measured height, weight, balancing on one foot, 3 meters
obstacle walking and required to fill in the holistic health lifestyle questionnaire. The results
presents the subjects should greatly improve the way of physical capability, nutrition, hygiene and the concept of illness prevention according to the
items of health life investigation. Furthermore, they should accumulate the regular
exercise habit especially in the improvement of muscular fitness.
Key Words: active aging, fitness, health promotion
P04 Effect of Methamphetamine on the Cell Proliferation of
RSC96 Schwann Cells
Jyun-Lin Zeng1, Meen-Woon Hsiao1, Jer-Yuh Liu2, 3, Sheng-Huang Chang4, and Jin-Ming Hwan1
1School of Applied Chemistry, Chung Shan Medical University, Taichung 2Graduate Institute of Cancer Biology, China Medical University, Taichung 3Center for Molecular Medicine, China Medical University,Taichung, and
4Department of Laboratory, Tsaotun Psychiatric Center, Ministry of Health and Welfare, Caotun, Nantou, Taiwan
happy540205@yahoo.com.tw
It is known that methamphetamine causes irreversible
abnormal changes in human brain structure and
accel- erated aging of brain cells. However
the mechanism is not well-understood. Schwann cells
within the peripheral nervous system are responsible
for myelination of axons, changes in their
action potentials, physiological changes such as growth
and differentiation, as well as protecting the
neurons and axons from damage. When Schwann
cells suffer pathological damage, it may cause
demyelination of axons, which in turn negatively
impact physiological functions and slow down transmission
of signals. Illnesses involving changes in the
peripheral nervous system are often tied to
pathological changes to Schwann cells, such as
spine injuries and responses to chemotherapy, diabetes
and chronic exposure to toxic heavy metals.
There is a possibility that the destruction of Schwann cells is related to pathological changes induced
by methamphetamine. This study investigated the damage induced by methamphetamine in
Schwann cells. RSC96 cells were treated with different concentrations of methamphetamine
(0, 0.7, 1.4, 2.1 and 2.8 mm), and observed a decrease in
cell growth rate (100%, 83%, 81%, 65% and 61% respec- tively).
Flow cytometry analysis showed that the cell cycles were observed to
be arrested at G0G1 phases, sug- gesting methamphetamine may cause cell
senescence in RSC96 cells.
Key Words: Schwann cells, methamphetamine, cell senescence
P05 Eight Probiotics Effective in Lowering Serum Lipids
Wen-Hung Wang1, Chih-Yin Hsu2, Jer-Yuh Liu3, 4, Wei-Jen Ting2, and Jin-Ming Hwan2
1Department of Food and Nutrition, Taichung Veterans General Hospital, Taichung 40705
2Department of Laboratory, Tsaotun Psychiatric Center, Ministry of Health and Welfare, Caotun, Nantou 542
3Graduate Institute of Cancer Biology, China Medical University, Taichung, and 4Center for Molecular Medicine, China Medical University,Taichung, Taiwan happy540205@yahoo.com.tw
Many types of Lactobacillus (LBs) have been suggested for use to lower the serum lipid, but this is not the case for all LBs. In the present study, eight probiotics (Lactobacillus acidophilus, Bifidobacterium bifidum, Lac- tobacillus casei, Bifidobacterium infantis, Bifidobacterium longum, Lactococcus lactis, Lactobacillus paracasei, Lactobacillus rhamnosus) including five LBs were selected and tested for their ability to lower the serum lipid level of male rats on a high-fat (12%) and high-cholesterol (0.2%) diet. This experiment included orally injecting
a set ratio of probiotics into rats which were given high lipid and high cholesterol diets. The
levels of serum high- density-lipoprotein-cholesterol (HDL-C) low-density-lipoprotein- cholesterol (LDL-C), Triglycerols (TG),
and total cholesterol (TC) were increased in the group given high
cholesterol and high fat (110 ± 12, 241 ± 97, 586 ± 218 and 546 ± 303, respectively), as compared to the control group (51 ± 8, 19 ± 5, 75 ± 15 and 104 ± 42, respec- tively). When treated with rosurastatin (rosuvastatin), the rats showed expected recovery of serum LDL-C, TG, and TC levels. When treated with various doses (200, 400, and 800 mg/kg) of the probiotics, the rats presented different levels of recovery of serum LDL-C, TG, and TC levels in a dose-dependent manner. Therefore we sug- gest that
the eight probiotics may have the potential function of lowering serum
lipid in rats.
Key Words: cholesterol, probiotics, triglycerols
P06 Effects of Breakfast Nutrition Education in Young Children
from Primary Caregivers
Yan-Jiun Huang and Chia-Xsun Wu
Department of Food and Nutrition, Providence University, Taichung, Taiwan
yjhuang@pu.edu.tw
Dietary habits in early childhood lasts the long influence until adulthood. Primary caregivers is the main factor for chileren’s establishment of dietary knowledge and behavior. The purpose of this study was to investigate the effects of the intervention of primary caregivers on young children’s breakfast behavior. Researchers Purposive sampling manner was used to recruit study objuct from Taichung kindergarten in age 5-6 years-old preschool children and their primary caregivers. Class as a unit, randomly divided into control group (N),
children and parents nutrition education group (CP), child nutrition education group
(C), and parents nutrition education group (P). Break- fast nutrition education
for young children was conducted 20-30 min, twice a week, for 2
weeks, same and deep- en content of newsletter for parents. Nutrition
questionnaire was given at the first and fourth week for both
young children and their caregivers. The results showed group C and CP
were significantly higher than the other two groups, and C was the highest, in
breakfast nutritional knowledge score at the fourth week. Group C and CP
were significantly higher than group P, group C was higher than group P, group
C was the highest, in breakfast eating behavior score at the fourth
week. Group C and CP had significantly higher than the first week in breakfast eating behavior. In conclusion,
face to face teaching was the most important way for young
children to learn. If parents do not have enough time to
interact with children would affect childre’s learning. The apparent importance of
young children’s nutritional education since knowledge is important backbone for the
establishment of behavior.
Key Words: young children, primary caregivers, breakfast,
nutrition education
P07 Sex Steroids Differentially Regulate Follicle-Stimulating Hormone (FSH)-Induced Progesterone Production in Ovarian
Granulosa Cells
Chi-Ying Hsieh, Hui-Ling Chen, and Jiuan-Jiuan Hwang
Institute of Physiology, School of Medicine, National Yang-Ming University, Taipei, Taiwan
jiuanh@ym.edu.tw
In females, ovary is the major organ for synthesis of sex steroids. This is under the control of pituitary-secreted FSH and ovarian local factors. Sex steroids, androgen and estrogen, have been proved to play important roles in folliculogenesis. Androgen also plays a role in the production of estrogen and progesterone at late follicle
stages, which is indispensable for the induction of ovulation. This study was to investigate the
differential effect of an- drogens and estrogen on FSH-induced progesterone synthesis
using primary culture of granulosa cells of PMSG- primed immature rats. First, androgens and estrogen dose-dependently enhanced FSH-induced progesterone production with various
potency (testosterone/T > androstenedione/AD > dihydrotestosterone/DHT > estradiol/E2). We further demonstrated that androgens modulate FSH-increased protein and mRNA level of key steroidogenic
proteins (StAR/Star, P450scc/Cyp11a1 and 3b-HSD/Hsd3b) and cholesterogenic protein (SR-BI/Scarb1) with different potency (AD ≈ T >> DHT), while E2
was without significant effect. Additionally, we found AD enhanced FSH-increased mRNA
levels of steroidogenic and cholesterogenic genes at 24 h but not at
4 h post-treatment. This could be in part due to androgen induction of its
receptor (AR) because treatment with AD in the absence or presence of
FSH upregulated AR at 24 h. In addition, AD enhancement of FSH-induced progesterone production
was inhibited by AR antagonist. In summary, present in vitro study
indicates in rat ovarian granulosa cells an- drogens could act through
AR to augment FSH-induced steroidogenic proteins and SR-BI, thus greatly augments
progesterone production.
Key Words: androgen, estrogen, FSH, ovarian granulosa
cell, progesterone, steroidogenesis
P08 Endothelin-1 Promotes MMP-2 Production and Migration in Human Fibrosarcoma Cells through PI3K/Akt Pathways
Kuan-Ming Chen1, Ming-Ju Chuang1, Wei-Ting Chao1, 2, and Min-Huan Wu2, 3
1Department of Life Science
2Life Science Research Center
and 3Physical Education Office, Tunghai University, Taichung, Taiwan
mhwu@thu.edu.tw
Tumor malignancy is associated with several cellular properties including proliferation and ability to metasta- size. Endothelin-1 (ET-1) the most potent vasoconstrictor plays a crucial role in migration and metastasis of human cancer cells. We found that treatment of human fibrosarcoma (HT1080 cells) with ET-1 increased migration and expression of
matrix metalloproteinase (MMP)-2. ET-1-mediated cell migration and MMP-2 expression were re-
duced by pretreatment with inhibitors of phosphatidylinositol 3-kinase (PI3K) and Akt,
as well as the NF-kB inhibitor and the IkB protease inhibitor. In addition, ET-1 treatment induced phosphorylation of PI3K, Akt, and resulted in increased NF-kB-luciferase activity that was inhibited by a specific inhibitor of PI3K, Akt, and NF-kB cascades. Taken together, these results suggest that ET-1 activated PI3K/AKT, which in turn
activated IKKa/b and NF-kB, resulting in increased MMP-2 expression and migration in human fibrosarcoma cells.
Key Words: endothelin-1, metastesis, fibrosarcoma, matrix
metalloproteinase
P09 Endothelin-1 Promotes Cells Motility through Enhancing Epithelial-Mesenchymal Transition in Colorectal Cancer
Bo-Ying Chen1, Pei-Shin Lee1, Wei-Ting Chao1, 2, and Min-Huan Wu2, 3
1Department of Life Science
2Life Science Research Center
and 3Physical Education Office, Tunghai University, Taichung, Taiwan
mhwu@thu.edu.tw
Despite considerable efforts to improve early detection and advances in chemotherapy, metastatic relapses re- main a major challenge in the management
of colorectal cancer. The endothelin receptor (ETR)/endothelin-1 (ET-1) axis has been shown to have a significant role in
colorectal cancer by promoting tumorigenesis. Here we show that the ET-1 pathway drives epithelial-to-mesenchymal transition (EMT) in colorectal cancer cells
by inducing migration and invasive phenotype, down-regulation of E-cadherin, increased levels
of N-cadherin, vimentin, Twist, and other mesenchymal markers. Pretreatment of colorectal
cancer cells with endothelin-receptors and AMP-activated pro- tein kinase (AMPKa) inhibitors abolished ET-1-promoted migration and increased the expression of N-cadherin and vimentin. On the other hand, ET-1 treatment demonstrably activated AMPKa and Twist signaling pathways. Taken together, our results indicate that ET-1 enhances the EMT markers and migratory ability
of colorectal cancer cells by increasing N-cadherin and vimentin expression via
the AMPKa, and Twist pathways.
Key Words: endothelin-1 (ET-1), colorectal cancer, epithelial-mesenchymal
transition, twist, migration
P10 Endothelin-1 Promotes IL-1beta Production in Skeletal Muscle Cells through PI3K/Akt Pathways
Chi-Cheng Lo1 and Min-Huan Wu2, 3
1Department of Life Science
2Life Science Research Center
3Physical Education Office, Tunghai University, Taichung, Taiwan
mhwu@thu.edu.tw
Endothelin-1 (ET-1) is a potent endogenous vasoconstrictor, which mediates
vascular wall cells proliferation, fibrosis, and inflammation through two types of
ET receptors (ETRs). IL-1beta is a multifunctional cytokine that plays a central role in both innate and acquired
immune responses. We investigated the signaling pathway in- volved in IL-1beta production stimulated by ET-1 in
cultured myoblasts. ET-1 caused concentration-dependent in- creases in IL-1beta production. Pretreatment with phosphatidylinositol 3-kinase
inhibitor, Akt inhibitor, NF-kappaB inhibitor (pyrrolidine dithiocarbamate, PDTC), and IkappaB protease
inhibitor (L-1-tosylamido-2-phenyl phenylethyl chloromethyl ketone, TPCK) also inhibited the potentiating action
of ET-1. ET-1 increased the PI3K, Akt phospho- rylation. Stimulation of myoblasts with ET-1 activated IKKalpha/beta,
p65 phosphorylation, and kappaB-luciferase activity. TNF-alpha mediated an increase of kappaB-luciferase
activity, which was inhibited by Ly294002, Akt in- hibitor, PDTC and
TPCK or PI3K, and Akt mutant. Our results suggest that ET-1 increased IL-1beta production in myoblasts via
PI3K/Akt and NF-kappaB signaling pathway.
Key Words: endothelin-1, inflammation, and skeletal muscle
P11 Endothelin-1 Promotes Lymphangiogenesis in Squamous Cell Carcinoma through Up-Regulated VEGF-C Expression
Yi-Ling Yu1, Bo-Cheng Chen4, Ya-Yu Huang1, Wei-Ting Chao1, 2, and Min-Huan Wu2, 3
1Department of Life Science
2Life Science Research Center
3Physical Education Office, Tunghai University, Taichung, Taiwan
4Department of Biological Science and Technology, China Medical University, Taichung, Taiwan
mhwu@thu.edu.tw
Oral cancer is one of the major
causes of mortality in humans and squamous
cell carcinoma is the most com- mon
type of oral cancer. Endothelin-1 (ET-1) has
been implicated in tumor Lymphangiogenesis and metastasis.
However, the relationship of ET-1 with vascular
endothelial growth factor C (VEGF-C) expression in human oral cells is mostly unknown. ET-1-mediated VEGF-C expression was assessed
by qPCR and Western blot analysis. The mechanisms of action of
ET-1 in different signaling pathways were studied using Western blotting. Knock-
down of proteins was achieved by transfection with siRNA. ET-1 increased
VEGF-C expression through P38, ERK, and sanil signaling cascades. Knockdown of ET-1 decreased VEGF-C expression and also abolished oral cancer conditional medium-mediated Lymphangiogenesis in
vitro. Taken together, these results indicate that ET-1 occurs through P38,
and ERK, which in turn activates snail, resulting in the activation of VEGF-C expression in human oral cancer.
Key Words: tongue squamous carcinoma cells (SCC4), endothelin-1
(ET-1), vascular endothelial growth factor C (VEGF-C), snail, P38, ERK
