OBJECTIVETo study the effect of basic fibroblast growth factor (bFGF) on the gene expression of syndecan-4 by human periodontal ligament cell (PDLC) in culture, and discuss the effect of bFGF on human PDLC proliferation and migration.

METHODS68 adolescent (12-18 years old) health premolar were collected, which were extracted for orthodontic reason. Human PDLC were cultured and stimulated by exogenous bFGF. After cultured 24, 48, 72h, gene expression of syndecan-4 was detected by SYBR green quantitative real time polymerase chain reaction.

RESULTSThe mRNA expression of syndecan-4 in 24 h group increased markedly than that in control group (P < 0.01), expecially in 1.0 ng x mL(-1) group. 1.0 ng x mL(-1) group in 48 h higher than that control group (P < 0.05). 1.0 ng x mL(-1) group in 72h compared with control group was lower (P < 0.05).

CONCLUSIONThe mRNA expression of syndecan-4 was increased by bFGF at the beginning, but the expression was decreased with the time. The expression of such changes may be one of the important factors which participate in the migration process of PDLC.

Cells, Cultured ; Fibroblast Growth Factor 2 ; Humans ; Periodontal Ligament ; RNA, Messenger ; Syndecan-4

BACKGROUND: Molecular mechanisms of natural killer (NK) cell development from hematopoietic stem cells (HSCs) have not been clearly elucidated, although the roles of some genes in NK cell development have been reported previously. Thus, searching for molecules and genes related NK cell developmental stage is important to understand the molecular events of NK cell development. METHODS: From our previous SAGE data-base, Gpnmb (Glycoprotein non-metastatic melanoma protein B) was selected for further analysis. We confirmed the level of mRNA and protein of Gpnmb through RT-PCR, quantitative PCR, and FACS analysis. Then we performed cell-based ELISA and FACS analysis, to know whether there are some molecules which can bind to Gpnmb. Using neutralizing antibody, we blocked the interaction between NK cells and OP9 cells, and checked IFN-gamma production by ELISA kit. RESULTS: Gpnmb expression was elevated during in vitro developmental stage and bound to OP9 cells, but not to NK precursor cells. In addition, we confirmed that the levels of Gpnmb were increased at NK precursor stage in vivo. We confirmed syndecan4 as a candidate of Gpnmb's binding molecule. When the interaction between NK cells and OP9 cells were inhibited in vitro, IFN-gamma production from NK cells were reduced. CONCLUSION: Based on these observations, it is concluded that Gpnmb has a potential role in NK cell development from HSCs.
Antibodies, Neutralizing ; Enzyme-Linked Immunosorbent Assay ; Hematopoietic Stem Cells ; Killer Cells, Natural ; Melanoma ; Polymerase Chain Reaction ; RNA, Messenger ; Syndecan-4

OBJECTIVETo investigate the effects of tumor necrosis factor-alpha(TNF-alpha) on syndecan-4 protein expression and proliferation of cultured human umbilical vein endothelial-like cells (HUVECs) in vitro.

METHODSHUVECs exposed to different concentrations of TNF-alpha(100, 20, 10, and 1 ng/ml) were cultured for 24 h and 36 h to observe their proliferation in comparison with the control group. The cell proliferation rate was determined by non-radioactive MTS/PES assay. The expression of syndecan-4 protein was evaluated by immunoblotting technique using anti-syndecan-4 antibody. Results The proliferation rate of the endothelial-like cells was 1.956-/+0.214 in the control group, and 2.154-/+0.250, 2.260-/+0.151, 2.118-/+0.205 and 2.106-/+0.136 in TNF-alpha-treated groups corresponding to TNF-alpha concentrations of 100, 20, 10 and 1 ng/ml at 24 h, respectively. It was shown that TNF-alpha significantly stimulated cell proliferation at the concentration above 1 ng/ml (P<0.05) as compared with the control group (P<0.05). The proliferation rate of the endothelial-like cell was 1.915-/+0.236 in the control group, and 2.067-/+0.328, 2.207-/+0.150, 2.052-/+0.126 and 2.051-/+0.180 in TNF-alpha-treated groups corresponding to TNF-alphaconcentrations of 100, 20, 10 and 1 ng/ml at 36 h, respectively. The expression of syndecan-4 protein was significantly enhanced by TNF-alpha.

CONCLUSIONSTNF-alpha can stimulate HUVEC proliferation, and expression of syndean-4 may represent an additional component of the pro-inflammatory, growth-stimulating pathways that are activated in response to changes in the vascular wall.

Cell Proliferation ; Cells, Cultured ; Endothelial Cells ; metabolism ; Humans ; Syndecan-4 ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology ; Umbilical Veins ; cytology

OBJECTIVE: To study the change and significance of serum pentraxin-3 (PTX-3) and syndecan-4 in children with chronic heart failure (CHF). METHODS: A total of 40 children with CHF who were admitted to the Department of Pediatrics of the First Affiliated Hospital of Zhengzhou University were enrolled as the heart failure group, and 30 children who underwent physical examination in the outpatient service during the same period of time were enrolled as the control group. The serum levels of PTX-3, syndecan-4, and N-terminal pro-brain natriuretic peptide (NT-proBNP) were compared between the two groups. RESULTS: The children with CHF had significant reductions in the serum levels of PTX-3, syndecan-4, and NT-proBNP after treatment. The levels of these markers in children with CHF were significantly higher than the control group before and after treatment ( CONCLUSIONS Serum PTX-3 and syndecan-4 may be involved in the development and progression of ventricular remodeling in children with CHF and may be used as markers for the diagnosis, cardiac function grading, and treatment outcome evaluation of children with heart failure.
Biomarkers ; Child ; Chronic Disease ; Heart Failure ; Humans ; Natriuretic Peptide, Brain ; Peptide Fragments ; Stroke Volume ; Syndecan-4 ; Ventricular Function, Left

PURPOSE: Transglutaminase type 2 (TG2) is an extracellular matrix crosslinking enzyme with a pivotal role in kidney fibrosis. We tested whether quantification of urinary TG2 may represent a noninvasive method to estimate the severity of kidney allograft fibrosis. METHODS: We prospectively collected urine specimens from 18 deceased donor kidney transplant recipients at 1-day, 7-day, 1-month, 3-month, and 6-month posttransplant. In addition, kidney allograft tissue specimens at 0-day and 6-month posttransplant were sampled to analyze the correlation of urinary TG2 and kidney allograft fibrosis. RESULTS: Thirteen recipients had increased interstitial fibrosis and tubular atrophy (IFTA) scores at the 6-month protocol biopsy (IFTA group). The mean level of urinary TG2 in the IFTA group was higher compared to that of 5 other recipients without IFTA (no IFTA group). Conversely, the mean level of urinary syndecan-4 in the IFTA group was lower than levels in patients without IFTA. In the IFTA group, double immunofluorescent staining revealed that TG2 intensity was significantly upregulated and colocalizations of TG2/heparin sulfate proteoglycan and nuclear syndecan-4 were prominent, usually around tubular structures. CONCLUSION: Urinary TG2 in early posttransplant periods is a potent biomarker for kidney allograft inflammation or fibrosis.
Allografts ; Atrophy ; Biomarkers ; Biopsy ; Extracellular Matrix ; Fibrosis ; Humans ; Inflammation ; Kidney Transplantation ; Kidney ; Methods ; Prospective Studies ; Proteoglycans ; Syndecan-4 ; Tissue Donors ; Transplant Recipients

Glycosaminoglycans are important structural components in the skin and exist as various proteoglycan forms, except hyaluronic acid. Heparan sulfate (HS), one of the glycosaminoglycans, is composed of repeated disaccharide units, which are glucuronic acids linked to an N-acetyl-glucosamine or its sulfated forms. To investigate acute ultraviolet (UV)-induced changes of HS and HS proteoglycans (HSPGs), changes in levels of HS and several HSPGs in male human buttock skin were examined by immunohistochemistry and real-time quantitative polymerase chain reaction (qPCR) after 2 minimal erythema doses (MED) of UV irradiation (each n = 4-7). HS staining revealed that 2 MED of UV irradiation increased its expression, and staining for perlecan, syndecan-1, syndecan-4, CD44v3, and CD44 showed that UV irradiation increased their protein levels. However, analysis by real-time qPCR showed that UV irradiation did not change mRNA levels of CD44 and agrin, and decreased perlecan and syndecan-4 mRNA levels, while increased syndecan-1 mRNA level. As HS-synthesizing or -degrading enzymes, exostosin-1 and heparanase mRNA levels were increased, but exostosin-2 was decreased by UV irradiation. UV-induced matrix metalloproteinase-1 expression was confirmed for proper experimental conditions. Acute UV irradiation increases HS and HSPG levels in human skin, but their increase may not be mediated through their transcriptional regulation.
Adult ; Agrin/genetics ; Antigens, CD44/genetics ; Base Sequence ; DNA Primers/genetics ; Gene Expression/radiation effects ; Glucuronidase/genetics ; Heparan Sulfate Proteoglycans/genetics/*metabolism ; Heparitin Sulfate/metabolism ; Humans ; Male ; Matrix Metalloproteinase 1/genetics ; N-Acetylglucosaminyltransferases/genetics ; RNA, Messenger/genetics/metabolism ; Skin/*metabolism/*radiation effects ; Skin Aging/genetics/physiology ; Syndecan-1/genetics ; Syndecan-4/genetics ; Ultraviolet Rays/*adverse effects ; Young Adult

BACKGROUND AND OBJECTIVES: Differentiation and de-differentiation of vascular smooth muscle cells (VSMCs) are important events in atherosclerosis and restenosis after angioplasty. MicroRNAs are considered a key regulator in cellular processes such as differentiation, proliferation, and apoptosis. Here, we report the role of new miR-18a-5p microRNA and its downstream target genes in VSMCs and in a carotid balloon injury model. MATERIALS AND METHODS: Expression of miR-18a-5p and its candidate genes was examined in VSMCs and in a carotid artery injury model by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and microRNA microarray analysis. VSMC differentiation marker genes including smooth muscle (SM) alpha-actin and SM22alpha were determined by Western blot, qRT-PCR, and a SM22alpha promoter study. Gene overexpression or knockdown was performed in VSMCs. RESULTS: miR-18a-5p was upregulated in the rat carotid artery at the early time after balloon injury. Transfection of the miR-18a-5p mimic promoted the VSMC differentiation markers SM alpha-actin and SM22alpha. In addition, miR-18a-5p expression was induced in differentiated VSMCs, whereas it decreased in de-differentiated VSMCs. We identified syndecan4 as a downstream target of miR-18-5p in VSMCs. Overexpression of syndecan4 decreased Smad2 expression, whereas knockdown of syndecan4 increased Smad2 expression in VSMCs. Finally, we showed that Smad2 induced the expression of VSMC differentiation marker genes in VSMCs. CONCLUSION: These results indicate that miR-18a-5p is involved in VSMC differentiation by targeting syndecan4.
Actins ; Angioplasty ; Animals ; Antigens, Differentiation ; Apoptosis ; Atherosclerosis ; Blotting, Western ; Carotid Arteries ; Carotid Artery Injuries ; Cell Differentiation* ; Microarray Analysis ; MicroRNAs* ; Muscle, Smooth ; Muscle, Smooth, Vascular* ; Polymerase Chain Reaction ; Rats ; Smad2 Protein ; Syndecan-4* ; Transfection

OBJECTIVETo investigate the effects of curcumin on the expression of syndecan-4 protein and p44/42 mitogen- activated protein kinase(MAPK) phosphorylation in rat vascular smooth muscle cells (VSMCs) induced by tumor necrosis factor-α (TNF-α) in vitro.

METHODSRat VSMCs cultured in vitro were stimulated for 24 h by 20 ng/ml TNF-α, 20 µmol/L curcumin, or 20 ng/ml TNF-α plus 20 µmol/lL curcumin. /assay was adopted to evaluate the proliferation of the VSMCs, and the expression of syndecan-4 protein and phosphorylated p44/42 MAPK were determined by Western blotting.

RESULTSCompared with the normal control cells, VSMCs exposed to TNF-α showed significantly enhanced proliferation (P/0.01). Curcumin treatment did not obviously affect the growth of otherwise untreated VSMCs(P>0.05), but could significantly suppress TNF-α-induced proliferation of VSMCs (P/0.01). TNF-α treatment also significantly increased the expression of syndecan-4 protein and phosphorylated p44/42 MAPK (P<0.01), which was markedly lowered by treatment with curcumin (P/0.01). Curcumin alone did not produce any obvious effects on the expression of syndecan-4 protein or phosphorylated p44/42 MAPK (P>0.05).

CONCLUSIONCurcumin can suppress the proliferation of rat VSMCs and lower the expression of syndecan-4 protein and phosphorylated p44/42 MAPK in TNF-α-induced VSMCs.

Animals ; Cells, Cultured ; Curcumin ; pharmacology ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Muscle, Smooth, Vascular ; drug effects ; metabolism ; Phosphorylation ; drug effects ; Rats ; Rats, Sprague-Dawley ; Syndecan-4 ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo investigate the effect of pravastatin on the proliferation of rat vascular smooth muscle cells (VSMCs) and expression of syndecan-4 protein induced by tumor necrosis factor-alpha (TNF-alpha).

METHODSVSMCs cultured in vitro were exposed to 20 ng/ml TNF-alpha, 10 micromol/ml pravastatin, 20 micromol/ml pravastatin, 10 micromol/ml pravastatin with 20 ng/ml TNF-alpha, or 20 micromol/ml pravastatin with 20 ng/ml TNF-alpha for 24 h. The proliferation of the VSMCs was determined by non-radioactive MTS/PMS assay and the expression of syndecan-4 protein was detected by Western blotting using anti-syndecan-4 antibody.

RESULTSCompared to the control group, TNF-alpha at 20 ng/ml significantly stimulated the proliferation of rat VSMCs (P<0.05). Pravastatin alone produced no obvious effect on VSMCs growth (P>0.05), but significantly inhibited TNF-alpha-induced VSMC proliferation (P<0.05). The expression of syndecan-4 protein in the VSMCs was significantly enhanced by 20 ng/ml TNF-alpha (P<0.01). Pravastatin alone did not affect the expression of syndecan-4 protein (P>0.05), but significantly inhibited TNF-alpha-induced enhancement of syndecan-4 protein expression (P<0.01).

CONCLUSIONPravastatin can inhibit the proliferation and syndean-4 protein expression in rat VSMCs induced by TNF-alpha in vitro.

Animals ; Anticholesteremic Agents ; pharmacology ; Aorta, Thoracic ; cytology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Pravastatin ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Syndecan-4 ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology