The study was to investigate the expression of VEGFA, VEGFC, angiopoietin-1, angiopoietin-2 and their receptors on yolk sac blood island, AGM region during gestation of 3th-12th weeks of human embryo. Human embryo contingently aborted at 3 - 12 weeks of gestation were collected with signed agreements of the pregnant women suffered from accidental abortions. The specimens were fixed by 4% paraformaldehyde and embedded by paraffin. 5 micro m serial sections were made. HE and immunohistochemistry method (SABC) and light-microscope were employed. The results showed that VEGFA and its receptors flt1/flk-1, VEGFC and its receptor flt-4, angiopoietin-2 and its receptor tie-2 proteins were expressed strongly and angiopoietin-1 was weakly expressed by hematopoietic cells and vascular endothelial cells of blood island at 21 and 25 days of gestation. In the 4th week of gestation, immuno-positive reaction of these factors and their receptors appeared in the aorta and mesonephros deposited in larger, rounded and nucleated cells which represented hematopoietic cells. Up to 7th week, positive hematopoietic cells in the regions were much abundant. The number of positive cells decreased at 8th week. Up to 12th week, almost all blood cells were immuno-negative. VEGFA, flt-1, flt-4, angiopoietin-1, angiopoietin-2 and Tie-2 protein were expressed mainly by gonad at 6 - 8 weeks, but it did not express VEGFC and flk-1. The immuno-reaction of the factors and their receptors could not detected in vascular endothelial cells during 3-12th weeks of gestation. It is concluded that hematopoietic cells and endothelial cells in blood island of yolk sac, mesonephros and dorsal aorta co-expressed some factors and their receptors in relation to vasculogenesis and hematopoiesis. Intraembryonic hematopoiesis began in the 4th week of gestation.
Angiopoietin-1 ; analysis ; Angiopoietin-2 ; analysis ; Embryo, Mammalian ; chemistry ; Extracellular Matrix Proteins ; analysis ; Humans ; Immunohistochemistry ; Receptor, TIE-2 ; analysis ; Receptors, Vascular Endothelial Growth Factor ; analysis ; Vascular Endothelial Growth Factor A ; analysis ; Vascular Endothelial Growth Factor C ; analysis ; Vascular Endothelial Growth Factor Receptor-1 ; Vascular Endothelial Growth Factor Receptor-2 ; analysis ; Vascular Endothelial Growth Factor Receptor-3 ; analysis ; Yolk Sac ; chemistry

OBJECTIVETo study the effect of dexamethasone on airway morphology and on the expression of angiopoietin-1 (Ang-1) and its tyrosine kinase receptor Tie-2 in the airway of asthmatic rats.

METHODSForty-five Sprague-Dawley rats were randomly divided into control, asthmatic, and dexamethasone-treated asthmatic groups. Asthma was induced by repeated sensitization and challenge with ovalbumin in the latter two groups. The dexamethasone intervention group received an intraperitonea injection of dexamethasone (2 mg/kg) before asthma challenge. Immunohistochemistry was used to measure the expression of Ang-1 and Tie-2 in the airway. Airway thickness was estimated by a computerized digital image analyzer.

RESULTSAirway thickness in the asthmatic group (33.9333+/-8.3791 micro m2/micro m) increased significantly compared with that in the control group (21.1333+/-2.7740 micro m2/micro m) (P<0.01). The dexamethasone intervention group also showed increased thickness of the airway (27.4000 +/- 4.6105 micro m2/micro m) compared with the control group (P<0.01), but the airway thickness in the dexamethasone intervention group was significantly reduced compared with that in the untreated asthmatic group (P<0.01). The expression of Ang-1 (103.9487+/-8.2914 vs 76.0320+/-3.7728; P<0.01) and Tie-2 (99.2307+/-8.1913 vs 75.3153+/-3.7321; P<0.01) in the airway increased significantly in the asthmatic group compared to controls. The expression of Ang-1 and Tie-2 in the airway of the dexamethasone intervention group (90.6180+/-5.2339 and 86.6633+/-3.7321, respectively) was statistically higher than that in the control group (P<0.01) but statistically lower than that in the untreated asthmatic group (P<0.01). Ang-1 and Tie-2 expression in the airway was positively correlated with the thickness of airway (r(Ang)-1=0.719r(Tie)-2=0.746P<0.01). There was also a positive correlation between Ang-1 and Tie-2 expression (r=0.742P<0.01).

CONCLUSIONSThe expression of Ang-1 and Tie-2 in the airway increased in asthmatic rats and was positively correlated with the thickness of the airway. Ang-1 and Tie-2 may participate in the process of airway remodeling in asthma. Dexamethasone can decrease the expression of Ang-1 and Tie-2 in the airway and relieve the changes of airway morphology.

Angiopoietin-1 ; analysis ; physiology ; Animals ; Asthma ; metabolism ; pathology ; Female ; Lung ; chemistry ; pathology ; Rats ; Rats, Sprague-Dawley ; Receptor, TIE-2 ; analysis ; physiology

The aim was to study the expression of VEGF-A, VEGF-C, angiopoietin-1, angiopoietin-2 and their receptors on development liver during gestation of weeks 3-12 of human embryo. Human embryo contingently aborted at 3-12 weeks of gestation were collected with signed agreements of the pregnant women suffered from accidental abortions. The specimens were fixed by 4% paraformaldehyde and embedded by paraffin. 5 microm serial sections were made. HE staining, immunohistochemistry method and light-microscope were employed. The results showed that at 4-5 weeks of development, liver was constituted by a few hepatic cords. Hematopoietic cell or blood cells were undetectable in the 4 week of gestation. A few cells which were larger, rounded and nucleared cells appeared and expressed VEGFA, flt-4 and Tie-2 proteins strongly in liver at 5 weeks of gestation. The number of these immuno-positive cells was highest in the 7th week and decreased at 11-12 weeks of gestation. These cells expressed flk-1 transiently in the 6th week. VEGF-C and flt-1 were expressed by hepatic cells from weeks 7 to 12 of gestation. The immuno-positive products were deposited in plasma of hepatic cells. Angiopoietin-1, angiopoietin-2 and Tie-2 were detectable on those cells which expressed VEGFA, flt-4 and Tie-2 from weeks 5 to 12 of gestation. The expression of angiopoietin-1 and angiopoietin-2 were weakly and Tie-2 was strongly. They were expressed weakly too by hepatic cells at 5 to 12 weeks of gestation. All factors and their receptors were undetectable on vascular endothelial cells at 4-12 weeks of gestation. It is concluded that the expression patterns of VEGF family on cells of liver are different before and after 7 weeks of gestation. The hematopoiesis in fetal liver may be related to development of hepatic cell.
Angiopoietin-1 ; analysis ; Angiopoietin-2 ; analysis ; Extracellular Matrix Proteins ; analysis ; Female ; Gestational Age ; Humans ; Immunohistochemistry ; Liver ; chemistry ; embryology ; Pregnancy ; Receptor, TIE-2 ; analysis ; Vascular Endothelial Growth Factor A ; analysis ; Vascular Endothelial Growth Factor C ; analysis ; Vascular Endothelial Growth Factor Receptor-1 ; Vascular Endothelial Growth Factor Receptor-3 ; analysis

PURPOSE: The purpose of this study was to evaluate the effect of alendronates on bone remodeling around titanium implant in the maxilla of rats. MATERIALS AND METHODS: The maxillary first molars were extracted and customized-titanium implants were placed immediately in thirty male Sprague-Dawley rats. The rats were divided into experimental (bisphosphonate) group and control group. At 4 weeks after implantation, the rats in the bisphosphonate group were subcutaneously injected with alendronate three times a week for 6 weeks where as the rats in control group were injected with saline. The rats were sacrificed at 1, 2, 3, 4, or 6 weeks after starting of injection and maxillary bones were collected subsequently. Alveolar bone remodeling around the implants were evaluated by radiographic and histologic analysis. Microarray analysis and immunohistomorphologic analysis were also performed on one rat, sacrificed at 6 weeks after starting of injection, from each group. Statistical analysis was performed using repeated measures analysis of variance and independent t test at a significance level of 5%. RESULTS: There was no statistically significant difference in the bone area (%) around implant between the bisphosphonate group and the control group. However, the amount of empty lacuna was significantly increased in the bisphosphonate group, especially in the rats sacrificed at 4 weeks after starting of injection compared to that of the corresponding control group. The bisphosphonate group showed the same level of TRAP positive cell count, osteocalcin and angiopoietin 1 as the control group. CONCLUSION: Alendronate may not decrease the amount of osteoclast. However, the significantly increased amount of empty lacuna in the bisphosphonate group may explain the suppression of bone remodeling in the bisphosphonate group.
Alendronate* ; Angiopoietin-1 ; Animals ; Bisphosphonate-Associated Osteonecrosis of the Jaw ; Bone Remodeling* ; Cell Count ; Humans ; Jaw ; Male ; Maxilla ; Microarray Analysis ; Molar ; Osteocalcin ; Osteoclasts ; Rats* ; Rats, Sprague-Dawley ; Titanium

OBJECTIVETo study the expression and significance of structural proteins, VEGF and Ang-1 in cavernous venous malformations of the body surface.

METHODSTissue samples came from 25 cases of cavernous venous malformations, 12 cases of normal moderate veins and 12 cases of normal small veins. Envision immunohistochemical stain was used to investigate the expression of IV collagen, fibronectin, laminin, VEGF and Ang-1. The results were analyzed semi-quantitatively.

RESULTSThe distribution of structural proteins in cavernous venous malformations is similar to moderate and small veins, but the expression in venous malformations is less obviously. VEGF expression in cavernous venous malformations and small veins is stronger obviously than moderate veins. Ang-1 expression in small veins is stronger remarkably than cavernous venous malformations and moderate veins.

CONCLUSIONThe abnormal expression of structural proteins may be an important factor in etiopathology and progress of cavernous venous malformations. There is disturbance of blood vessel remodelling in the sinusoid of cavernous venous malformations, with which the less expression of Ang-1 may be related.

Angiopoietin-1 ; metabolism ; Collagen Type IV ; metabolism ; Fibronectins ; analysis ; Hemangioma, Cavernous ; metabolism ; Humans ; Laminin ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Veins ; abnormalities ; metabolism

BACKGROUNDAngiopoietin-1 (Ang-1) is an endothelial-specific growth factor that can promote angiogenesis. Studies demonstrated that Ang-1 can inhibit apoptosis of umbilical endothelial cells, but so far little is known about its effects on apoptosis of microvascular endothelial cells. With the apoptotic model of murine-cerebral-derived microvascular endothelial cells (bEnd.3) induced by serum-free culture, we attempted to clarify the molecular mechanism of bEnd.3 apoptosis, particularly its relation to cytochrome C (Cyt C).

METHODSThe cultured microvascular endothelial cell strain, bEnd.3 cell, was employed. An apoptotic model of bEnd.3 was established by serum-free culture. Flow cytometry after Annexin labeling and PI staining were used to assess the apoptotic effects of Ang-1 on bEnd.3, and the expression of Bax/Bcl-2, caspase 8, caspase 3, and Cyt C were detected with Western blotting and ELISA.

RESULTSThe apoptotic rate of bEnd.3 cells after stimulation with Ang-1 (100 ng/L) in serum-free medium was significantly higher than that in control group. Ang-1 inhibited early-stage apoptosis more than late-stage apoptosis provided by propidium iodide (PI) and AnnexinV double staining. The inhibition of Ang-1 on bEnd.3 cell apoptosis was strengthened with the increase in concentration (0 - 400 ng/ml). Ang-1 could decrease the expression of Bax, caspase3 and 8, and increase that of Bcl-2. The results of ELISA indicated that Ang-1 significantly decreased CytC content in cytoplasm and increase that in mitochondria.

CONCLUSIONSAng-1 could inhibit bEnd.3 apoptosis induced by serum-free medium culture. The apoptosis was associated with decreased Bax expression, increased Bcl-2 expression, which result in Cyt C transferring from mitochondria to cytoplasm, and then caspases activation are reduced and cell apoptosis is suppressed.

Angiopoietin-1 ; pharmacology ; Animals ; Annexin A5 ; analysis ; Apoptosis ; drug effects ; Caspase 3 ; Caspase 8 ; Caspases ; analysis ; Cells, Cultured ; Cytochromes c ; physiology ; Endothelial Cells ; cytology ; drug effects ; Mice ; Microcirculation ; drug effects ; bcl-2-Associated X Protein ; analysis

BACKGROUNDEndothelial progenitor cells (EPCs) have been used in both experimental studies and clinical treatments of limb ischemia, as well as in the construction of engineered vascular tissue. The objective of this study was to investigate the effects of transplanted bone marrow-derived EPCs on the vein microenvironment in a rat model of chronic vein thrombosis.

METHODSMononuclear cells were isolated from the bone marrow of immature rats by density gradient centrifugation, cultured, and then transplanted into experimentally induced thrombi into inferior vena cava through the femoral vein. Vascular endothelial growth factor (VEGF), angiopoietin-1 (ANG-1) and monocyte chemotactic protein-1 (MCP-1) mRNA and protein expression levels were measured by real-time quantitative polymerase chain reaction and Western blotting of thrombi and adjacent caval walls 28 days post-transplantation.

RESULTSLevels of VEGF, ANG-1, and MCP-1 mRNA in EPC-transplanted thrombi were 100%, 230.7%, and 212.5% of levels detected in the sham-operated group (P < 0.01), and 99.9%, 215.4%, and 177.8% of levels detected in the experimental control group (P < 0.01). VEGF, ANG-1 and MCP-1 protein levels exhibited a similar trend.

CONCLUSIONSTransplanted bone marrow-derived EPCs appear to alter the vein microenvironment in experimentally induced chronic vein thrombosis by upregulating cytokines associated with thrombic organization and recanalization.

Angiopoietin-1 ; analysis ; genetics ; Animals ; Bone Marrow Cells ; cytology ; Chemokine CCL2 ; analysis ; genetics ; Chronic Disease ; Disease Models, Animal ; Endothelial Cells ; cytology ; Fluorescent Antibody Technique ; Immunohistochemistry ; RNA, Messenger ; analysis ; Rats ; Stem Cell Transplantation ; Vascular Endothelial Growth Factor A ; analysis ; genetics ; Venous Thrombosis ; therapy

Cartilage oligomeric matrix protein-angiopoietin-1 (COMP-Ang1) is an angiogenic factor for vascular angiogenesis. The aim was to investigate the effect of an intracavernosal injection of COMP-Ang1 on cavernosal angiogenesis in a diabetic rat model. Male Otsuka Long-Evans Tokushima Fatty (OLETF) rats made up the experimental group (1 yr old) and Long-Evans Tokushima Otsuka (LETO) rats made up the control group. The experimental group was divided into vehicle only, 10 microg COMP-Ang1, and 20 microg COMP-Ang1. COMP-Ang1 was injected into the corpus cavernosum of the penis. After 4 weeks, the penile tissues of the rats were obtained for immunohistochemistry and Western blot analysis. The immunoreactivity of PECAM-1 and VEGF was increased in the COMP-Ang1 group compared with the vehicle only group. Moreover, the expression of PECAM-1 and VEGF was notably augmented in the 20 microg Comp Ang-1 group. In the immunoblotting study, the expression of PECAM-1 and VEGF protein was significantly less in the OLEFT rats than in the control LETO rats. However, this expression was restored to control level after intracavernosal injection of COMP-Ang1. These results show that an intracavernosal injection of COMP-Ang1 enhances cavernous angiogenesis by structurally reinforcing the cavernosal endothelium.
Angiopoietin-1/genetics/*metabolism ; Animals ; Antigens, CD31/metabolism ; Blood Glucose/analysis ; Blotting, Western ; Body Weight ; Cartilage Oligomeric Matrix Protein/genetics/*metabolism ; Diabetes Mellitus, Experimental/*pathology ; Immunohistochemistry ; Male ; Neovascularization, Physiologic/*drug effects ; Penis/metabolism/pathology ; Rats ; Rats, Long-Evans ; Recombinant Fusion Proteins/biosynthesis/genetics/*pharmacology ; Vascular Endothelial Growth Factor A/metabolism

Previous studies suggested that polymorphisms of proinflammatory cytokine genes are important host genetic factors in Helicobacter pylori infection. The present study evaluated whether IL-8-251 polymorphism affected H. pylori eradication rate and to investigate the effect of H. pylori eradication on angiogenesis and the inflammatory process according to the IL-8-251 polymorphism. A total of 250 H. pylori-positive patients treated by endoscopic resection of the gastric neoplasm were classified into 3 groups (134 H. pylori-eradicated group, 19 H. pylori-eradication failure group, and 97 H. pylori-infected group). H. pylori status, histology, and angiogenic factor levels were evaluated at baseline, 6 months, and 18 months. H. pylori eradication rate was 92.9% in AA genotype, 85.7% in AT genotype and 88.4% in TT genotype (P value = 0.731). Elevated IL-8 and matrix metalloproteinase-9 concentrations in H. pylori-infected gastric mucosa were reversible by successful eradication of H. pylori, independent of the IL-8-251 polymorphism. It is suggested that elevated IL-8 and MMP-9 concentrations in H. pylori-infected gastric mucosa are altered significantly after successful eradication and these conditions continue for 18 months. However, IL-8-251 polymorphism does not affect H. pylori eradication rate and the sequential changes of related angiogenic factors after H. pylori eradication in Koreans.
Aged ; Alleles ; Angiopoietin-1/analysis ; Anti-Bacterial Agents/therapeutic use ; Anti-Inflammatory Agents, Non-Steroidal/therapeutic use ; Asian Continental Ancestry Group/*genetics ; Female ; Gastric Mucosa/metabolism/pathology ; Genotype ; Helicobacter Infections/*drug therapy ; *Helicobacter pylori ; Humans ; Interleukin-8/analysis/*genetics ; Male ; Matrix Metalloproteinase 9/analysis ; Middle Aged ; *Polymorphism, Single Nucleotide ; Proton Pump Inhibitors/therapeutic use ; Republic of Korea ; Retrospective Studies ; Stomach Neoplasms/pathology/surgery ; Time Factors ; Vascular Endothelial Growth Factor A/analysis