Thrombospondin-1 (TSP-1), a multifunctional extracellular matrix protein, inhibits neovascularization and is implicated in the regression of tumor growth and metastasis. We found that the synthesis of TSP-1 in porcine aortic endothelial (PAE) cells was decreased in a dose-dependent manner by phorbol 12-myristate 13-acetate (PMA) treatment in porcine aortic endothelial (PAE) cells. In this study, a responsive site on the TSP-1 promotor affected by PMA treatment in PAE was characterized. The level of TSP-1 mRNA was also decreased by PMA after 1 h and persisted that way for at least 24 h. PMA treatment and c-Jun overexpression suppressed the transcription of TSP-1 promotor-luciferase reporter gene. A deletion between -767 and -657 on the TSP-1 promotor neutralized the PMA-induced down-regulation. In addition, oligo a (-767 approximately -723) was responsive to PMA-induced repression, while oligo b (-734 approximately -689) and c (-700 approximately -656) was not. Electrophoretic mobility shift assays showed that this PMA responsive element specifically bound a nuclear protein and that the binding activity was diminished by PMA treatment in PAE cells but not in Hep 3B cells. In supershift assay, potential regulatory elements in this region, SP1 and GATA-1, were not responsive to the inhibition of TSP-1 expression by PMA. Our results suggest that the repression of TSP-1 synthesis by PMA is mediated by blocking a particular unknown nuclear protein binding to the responsive site (-767 approximately -735), which is regulated by c-Jun.
Animal ; Aorta/cytology ; Cell Line ; Down-Regulation (Physiology) ; Endothelium, Vascular/drug effects* ; Endothelium, Vascular/cytology ; Promoter Regions (Genetics)* ; Proto-Oncogene Proteins c-jun/metabolism ; Response Elements* ; Swine ; Tetradecanoylphorbol Acetate/pharmacology* ; Thrombospondin 1/genetics* ; Thrombospondin 1/biosynthesis

Expression of thrombospondin-1 (TSP-1), which is a known inhibitor of tumor growth and angiogenesis, is reciprocally regulated by positive regulators, such as VEGF. Additionally, trichostatin A (TSA) suppresses tumor progression by altering VEGF levels and VEGF-mediated signaling. Thus, understanding TSA-regulated TSP-1 expression and the effects of altered TSP-1 levels might provide insights into the mechanism of action of TSA in anti-tumorigenesis, and provide an approach to cancer therapy. Here, we examined the effect of TSA on TSP-1 expression, and the effects of TSA-induced TSP-1 on cell motility and angiogenesis, in HeLa and bovine aortic endothelial cells. TSA remarkably increased TSP-1 expression at the mRNA and protein levels, by controlling the TSP-1 promoter activity. Both TSA and exogenous TSP-1 reduced cell migration and capillary-like tube formation and these activities were confirmed by blocking TSP-1 with its neutralizing antibody and small-interfering RNA. Our results suggest that TSP-1 is a potent mediator of TSA-induced anti- angiogenesis.
Angiogenesis Inhibitors/*pharmacology ; Animals ; Cattle ; Cell Line ; Cell Movement/*drug effects ; Endothelial Cells/drug effects/*physiology ; Humans ; Hydroxamic Acids/*pharmacology ; Neovascularization, Pathologic/metabolism/prevention & control ; Neovascularization, Physiologic/*drug effects ; RNA, Messenger/biosynthesis ; RNA, Small Interfering/*genetics ; Thrombospondin 1/*biosynthesis/genetics/pharmacology

Thrombospondin-1 (TSP-1), a multifunctional protein that is able to function as a negative regulator of solid tumor progression and angiogenesis, is normally present at a very low level but rapidly elevated in pathological tissues. To understand the cellular regulation of TSP-1 expression, the mode of it's expression in Hep3B, SK-HEP-1, and porcine aortic endothelial (PAE) cells was examined in the presence of all-trans retinoic acid (ATRA), interleukin-6 (IL-6), interferon-gamma (IFN-gamma), or phorbol 12-myristate 13-acetate (PMA). ATRA or IL-6 induced a dose-dependent increase of TSP-1 protein and mRNA levels in PAE cells, while they negatively regulated TSP-1 expression in the Hep3B and SK-HEP-1 cells. In contrast, PMA showed just the opposite effects on the TSP-1 expression in the same cells. IFN-gamma had little effect on TSP-1 level in Hep3B and PAE cells. The TSP-1 expression in SK-HEP-1 cells by these agents showed a close resemblance to that of liver cells rather than that of the endothelial cell line. Possible TSP-1 promoter-mediated responses by ATRA, IL-6, IFN-gamma, or PMA in Hep3B and PAE cells examined with luciferase activity of TSP-LUC reporter plasmid showed that levels of TSP-1 promoter activity were lower than that of the expressed TSP-1 protein and mRNA levels. Transfection of c-Jun and/or RARalpha expression vectors into Hep3B and PAE cells resulted in the enhanced TSP-1 promoter activity as well as the increments of of its protein and mRNA level. These results suggest that regulatory agents-induced TSP-1 expression may be attributed to mRNA stability and/or translational activation in concert with transcriptional activation and TSP-1 expression may be independently controlled via each signal pathway stimulated by PMA or ATRA.
Animal ; Cell Line ; Endothelium, Vascular/cytology/drug effects/metabolism ; *Gene Expression Regulation/drug effects ; Genes, Reporter ; Genes, jun ; Human ; Immunoblotting ; Interferon Type II/pharmacology ; Interleukin-6/pharmacology ; *Promoter Regions (Genetics) ; Proto-Oncogene Proteins c-jun/genetics/metabolism ; Receptors, Retinoic Acid/genetics/metabolism ; Recombinant Fusion Proteins/metabolism ; Tetradecanoylphorbol Acetate/pharmacology ; Thrombospondin 1/*genetics/metabolism ; Transcription, Genetic ; Tretinoin/pharmacology

PURPOSE: The aim of this study is to find the prevalence of obesity, the serum lipid levels and the age at menarche in adolescent female athletes and to examine the effects of exercise in adolescent stage. METHODS: With a questionnaire regarding their age at menarche, physical measurement, body fat, and blood samples of the serum lipid levels to evaluate the hyperlipidemia were obtained from adolescent female athletes(n=107) and general adolescent students(n=650) who didn't exercise at regular intervals, aged 12 to 18 years. RESULTS: The mean weight in the athletes' group was 53.3+/-7.3 kg, which was similar with 54.3+/-8.0 kg in the control group. The mean height in the athletes' group was 161.4+/-5.4 cm, which was taller than 158.9+/-5.3 cm in the control group. The prevalence of obesity by obesity index, body fat, and BMI in the athletes' group were significantly lower than in control group. There was no significant difference in age at menarche between two groups(12.6+/-1.3, 12.9+/-1.2). The levels of cholesterol, LDL cholesterol, and HDL cholesterol in the athletes' group were higher than in the control group. The levels of triglyceride in the athletes' group was lower than in control group. CONCLUSION: These data suggest the importance of exercise in adolescents for the prevention of obesity since it may reduce body fat and increase the height. There was no negative effect of exercise on the age at menarche. We think that more controlled assessment of nutrition, diet habit, hormonal effect and height are warranted to find the correlation with hyperlipidemia and exercise at the adolescent stage.
Adipose Tissue ; Adolescent* ; Athletes* ; Cholesterol ; Cholesterol, HDL ; Cholesterol, LDL ; Female ; Female* ; Food Habits ; Humans ; Hyperlipidemias ; Menarche* ; Obesity* ; Prevalence* ; Surveys and Questionnaires ; Triglycerides

Nephrotic syndrome occurs infrequentiy prior to the second year of life. When nephrotic syndrome does develop during the first year, the course differs from that of older children with nephrotic sydrome, being characterized by an extremely poor prognosis and an almost complete refractoriness to therapy. Despite its low incidence congenital nephrotic syndrome is important, not only because of the severity and the disorder itself but also because the occurrence of nephrotic syndrome in this age group rasies question regarding the etiology of the disease. We experienced one case of congenital nephrotic syndrome which was confirmed by autopsy. The patient was born as a premature infant with body weight 1,400gm and 37weeks of gestational age, to a toxemic mother, gravida 3 and parity 3 in Obsteric Department of Seoul Red Cross Hospital. The patient had an uncomplicated nursery staying even though routine weekly urinalysis did show up various degree of proteinuria and microscopic hematuria without any obvious edema till the age of 50 days with body weight 2,400gm on the discharge. He was lost to be followed at Out Patient Department untill the age of 4months when he was brought to admission because of respiratory distress in generalized edematous state. He died at 7 months of age following progressive down-hill cours, despite treatment with prednisolone and cyclophosphamide for 2 months. At autopsy, almost all of the glomeruli (99%) were sclerotic with occassional creascent formation and tubules showed mircocystic dilataions. It is considered that this case was the first one which was presented on literature in Korea.
Autopsy ; Body Weight ; Child ; Cyclophosphamide ; Edema ; Female ; Gestational Age ; Hematuria ; Humans ; Incidence ; Infant, Newborn ; Infant, Premature ; Korea ; Mothers ; Nephrotic Syndrome* ; Nurseries ; Parity ; Prednisolone ; Prognosis ; Proteinuria ; Red Cross ; Seoul ; Urinalysis

Purpose: This study compared clinical and radiologic differences between cystic biliary atresia (cBA) and choledochal cyst (CC) type Ia/b. Methods: Infants (≤12 months old) who were diagnosed with cBA or CC type Ia/b from 2005 to 2019 were retrospectively reviewed. Imaging features on preoperative ultrasonography (US) and magnetic resonance imaging (MRI) were compared between the cBA and CC groups. Logistic regression and area under the receiver operating characteristic curve (AUC) analyses were performed for the diagnosis of cBA. Changes in cyst size were also evaluated when prenatal US exams were available. Results: Ten patients (5.5% of biliary atresia cases) with cBA (median age, 48 days) and 11 infants with CC type Ia/b (Ia:Ib=10:1; median age, 20 days) were included. Triangular cord thickness on US (cutoff, 4 mm) showed 100% sensitivity and 90.9% specificity (AUC, 0.964; 95% confidence interval [CI], 0.779 to 1.000) and cyst size on MRI (cutoff, 2.2 cm) had 70% sensitivity and 100% specificity (AUC, 0.900; 95% CI, 0.690 to 0.987) for diagnosing cBA. Gallbladder mucosal irregularity on US and an invisible distal common bile duct on MRI were only seen in the cBA group (10 of 10). Only the CC group showed prenatal cysts exceeding 1 cm with postnatal enlargement. Conclusion Small cyst size (<1 cm) on prenatal US, triangular cord thickening (≥4 mm) and gallbladder mucosal irregularity on postnatal US, and small cyst size (≤2.2 cm) and an invisible distal common bile duct on MRI can discriminate cBA from CC type Ia/b in infancy.

BACKGROUND: Hyperhomocysteinemia is an independent risk factor for cardiovascular diseases. Previous reports showed that homocysteine damages mitochondrial gene expression, function and structure. In recent years, homocysteine and mitochondrial DNA (mtDNA) content are reported to relate with insulin resistance. The aim of this study is to investigate the correlation of plasma homocysteine level and mitochondrial DNA content in peripheral blood. METHODS: The mtDNA content, homocysteine and insulin resistance parameters were measured in healthy women (n=60). Plasma homocysteine level was measured by ion-exchange chromatography method and the mtDNA content in peripheral blood was measured by real time PCR method using ABI Prism 7700 machine. RESULTS: Significant correlation was found between homocysteine and mtDNA content (r=-0.507, p<0.05). Homocysteine was correlated with age (r=0.397), cholesterol (r=0.327), LDL-cholesterol (r=0.318), apolipoprotein B (r=0.387), HbA1c (r=0.274) positively and folate (r=-0.262), apolipoprotein A1 (r=-0.293), VO2max (r=-0.332) negatively. Mitochondrial DNA content was correlated with age (r=-0.535), BMI (r=-0.397), cholesterol (r=-0.340), LDL-cholesterol (r=-0.319), apolipoprotein B (r=-0.367) negatively and apolipoprotein A1 (r=0.346), lactate (r=0.307), VO2max (r=0.308) positively. All correlations were statistically significant(p<0.05). CONCLUSION: In this study, plasma homocysteine level was related with mitochondrial DNA content negatively and these two factors are estimated to be concerned with insulin resistance.
Apolipoprotein A-I ; Apolipoproteins ; Cardiovascular Diseases ; Cholesterol ; Chromatography, Ion Exchange ; DNA, Mitochondrial* ; Female ; Folic Acid ; Genes, Mitochondrial ; Homocysteine* ; Humans ; Hyperhomocysteinemia ; Insulin Resistance ; Lactic Acid ; Plasma* ; Real-Time Polymerase Chain Reaction ; Risk Factors

OBJECTIVES: The purpose of this study was to evaluate the characteristics of psychopathology, adverse drug effects, and subjective response to drugs that have a significant impact on the subjective quality of life in schizophrenic patients receiving atypical antipsychotics. METHODS: One hundred and one schizophrenic patients, who were receiving maintenance treatment with atypical antipsychotics, were evaluated. Subjective quality of life was assessed using the standardized Korean modification of a self-rating scale to measure subjective well-being under neuroleptics (KmSWN). Patients' psychopathology was evaluated using the Positive and Negative Syndrome Scale. Adverse effects and subjective response to drug were evaluated using the Liverpool University Neuroleptic Side Effect Rating Scale and the Drug Attitude Inventory-10, respectively. Correlation analysis and stepwise multiple regression analysis were conducted. RESULTS: In psychopathology, the severity of depression and anxiety showed the most significant correlation with the score of KmSWN. In adverse drug effects, the severity of psychic side effect and extrapyramidal side effect showed the most significant correlation with the score of KmSWN. Regarding subjective response to drug, significant correlation was observed between the severity of subjective negative response and the score of KmSWN. Stepwise multiple regression analysis revealed that psychic side effect, extrapyramidal side effect, and depression contributed significantly to the total score of KmSWN. These variables accounted for 59.7% of the total variance. CONCLUSION: The results of the present study suggest that psychic side effect, extrapyramidal side effect and depressive symptom are the clinical characteristics that are significantly associated with the subjective quality of life. An effective management strategy for these variables should be established in developing a treatment program to enhance the quality of life of patients with schizophrenia.
Antipsychotic Agents* ; Anxiety ; Depression ; Humans ; Psychopathology* ; Quality of Life* ; Schizophrenia

BACKGROUND: The incidence of atherosclerosis is well correlated with the progression of type 2 diabetes mellitus. High plasma glucose in uncontrolled diabetic patients evokes many vascular complications such as atherosclerosis. Specifically, high glucose was reported to induce thrombospondin-1 (TSP-1), which activates matrix metalloproteinase-2 (MMP-2) and leads to the invasion of vascular smooth muscle cells (VSMCs) into the intima. Catechins with antioxidant effects are known to inhibit MMP-2 activity. Therefore, this study was aimed at revealing the effect of epicatechin, one of catechins, on high glucose-induced TSP-1 and the invasiveness of VSMCs. METHODS: VSMCs were primarily isolated from Sprague-Dawley rat aorta. The VSMCs were incubated with different doses (30, 100 and 300 micrometer) of epicatechin under high glucose concentration (30 mM). The TSP-1 protein and mRNA expressions were analyzed by performing Western blotting and Northern blot analyses, respectively. RT-PCR was performed to observe the MMP-2 mRNA expression. Gelatin zymography was performed for the measurement of MMP-2 activity. Invasion assays were performed to evaluate the invasiveness of VSMCs. RESULTS: Epicatechin inhibited the high glucose-induced TSP-1 expression and the MMP-2 activity in a dose-dependent manner. Also, epicatechin inhibited the high glucose-induced invasiveness of VSMCs across the matrix barrier in a dose-dependent fashion. CONCLUSION: Collectively, epicatechin may prevent the high glucose-induced proliferation and invasion of VSMCs by inhibiting the TSP-1 expression and the MMP-2 activity. Therefore, epicatechin appears to play a protective role in the development of atherosclerosis.
Animals ; Antioxidants ; Aorta ; Atherosclerosis ; Blood Glucose ; Blotting, Northern ; Blotting, Western ; Catechin* ; Diabetes Mellitus, Type 2 ; Gelatin ; Glucose ; Humans ; Incidence ; Matrix Metalloproteinase 2 ; Muscle, Smooth, Vascular* ; Rats* ; Rats, Sprague-Dawley ; RNA, Messenger ; Thrombospondin 1*

BACKGROUND: The incidence of atherosclerosis is well correlated with the progression of type 2 diabetes mellitus. High plasma glucose in uncontrolled diabetic patients evokes many vascular complications such as atherosclerosis. Specifically, high glucose was reported to induce thrombospondin-1 (TSP-1), which activates matrix metalloproteinase-2 (MMP-2) and leads to the invasion of vascular smooth muscle cells (VSMCs) into the intima. Catechins with antioxidant effects are known to inhibit MMP-2 activity. Therefore, this study was aimed at revealing the effect of epicatechin, one of catechins, on high glucose-induced TSP-1 and the invasiveness of VSMCs. METHODS: VSMCs were primarily isolated from Sprague-Dawley rat aorta. The VSMCs were incubated with different doses (30, 100 and 300 micrometer) of epicatechin under high glucose concentration (30 mM). The TSP-1 protein and mRNA expressions were analyzed by performing Western blotting and Northern blot analyses, respectively. RT-PCR was performed to observe the MMP-2 mRNA expression. Gelatin zymography was performed for the measurement of MMP-2 activity. Invasion assays were performed to evaluate the invasiveness of VSMCs. RESULTS: Epicatechin inhibited the high glucose-induced TSP-1 expression and the MMP-2 activity in a dose-dependent manner. Also, epicatechin inhibited the high glucose-induced invasiveness of VSMCs across the matrix barrier in a dose-dependent fashion. CONCLUSION: Collectively, epicatechin may prevent the high glucose-induced proliferation and invasion of VSMCs by inhibiting the TSP-1 expression and the MMP-2 activity. Therefore, epicatechin appears to play a protective role in the development of atherosclerosis.
Animals ; Antioxidants ; Aorta ; Atherosclerosis ; Blood Glucose ; Blotting, Northern ; Blotting, Western ; Catechin* ; Diabetes Mellitus, Type 2 ; Gelatin ; Glucose ; Humans ; Incidence ; Matrix Metalloproteinase 2 ; Muscle, Smooth, Vascular* ; Rats* ; Rats, Sprague-Dawley ; RNA, Messenger ; Thrombospondin 1*