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1.
Article in English | WPRIM | ID: wpr-301076

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the effect of Xuefu Zhuyu Capsule ()-containing serum (XFZY-CS) on EphB4/ephrinB2 and its reverse signal in human microvascular endothelial cell-1 (HMEC-1).</p><p><b>METHODS</b>XFZY-CS and the blank control serum were collected. HMEC-1 cells were randomly assigned to 6 groups including the concentration 1.25%, 2.5%, and 5% XFZY-CS groups and their blank serum control ones. The angiogenesis effect of XFZY-CS was tested with an in vitro tube formation assay and the best condition of pro-angiogenesis was determined. The effect of XFZY-CS on EphB4/ephrinB2 and the reverse signal were determined by Western blot and real-time quantitative polymerase chain reaction, respectively; we also confifirmed the results through activating and inhibiting the reverse signal by EphB4/fc and pyrophosphatase/ phosphodiesterase2 (PP2).</p><p><b>RESULTS</b>XFZY-CS promoted angiogenesis at the concentration of 2.5% corresponding serum after being cultured for 48 h, while inhibited angiogenesis at the concentration of 5% after culturing for 48 and 72 h. Under the 2.5% serum concentration, XFZY up-regulated the expression of EphB4-mRNA at 12 h (P<0.05), and down-regulates its expression at 24 h (P<0.01). Protein expression of EphB4 was apparently up-regulated at 12 h and down-regulated at 24 h. The phosphorylation of ephrinB2 increased at 9 h (P<0.05). In addition, 2.5% XFZY-CS played a similar role as the reverse signaling activator EphB4/Fc ranging from 0.5 to 5 μg/mL (P>0.05). XFZY-CS also reduced the inhibitive effect of PP2 in limited periods.</p><p><b>CONCLUSIONS</b>EphB4/ephrinB2 was the upstream signal in the process of angiogenesis and its reverse signaling was responsible for XFZY's effect on promoting angiogenesis.</p>


Subject(s)
Adult , Humans , Male , Middle Aged , Young Adult , Capsules , Drugs, Chinese Herbal , Pharmacology , Endothelial Cells , Metabolism , Ephrin-B2 , Metabolism , Gene Expression Regulation , Microvessels , Pathology , Neovascularization, Physiologic , Genetics , Phosphoric Diester Hydrolases , Metabolism , RNA, Messenger , Genetics , Metabolism , Receptor, EphB4 , Genetics , Metabolism , Serum , Metabolism , Time Factors
2.
Chinese Journal of Stomatology ; (12): 151-156, 2015.
Article in Chinese | WPRIM | ID: wpr-360430

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of local delivery of delta12-prostaglandinJ2-loaded poly (lactic-co-glycolic acid) (Δ(12)-PGJ2-NC) on growth factors expression and bone formation.</p><p><b>METHODS</b>Δ(12)-PGJ2-NC was prepared by the emulsion solvent diffusion method. The physical and chemical properties of the nanoparticles were evaluated by particle size analysis, transmission electron microscopy, drug-loading ratio and the in vitro release study. Then standardized transcortical defect (5.0 mm × 1.5 mm) was conducted in the femur of 48 male Wistar rats which were randomly divided into four groups (n = 12), S, K, F, and N. Thirty microliter of saline (S), unloaded nanoparticles (K), Δ(12)-PGJ2 (F) and Δ(12)-PGJ2-NC(N) in a collagen vehicle were delivered inside a titanium chamber fixed over the defect. Then, four subgroups were randomly divided in each group named as D3, D7, D14, and D28 (n = 3) according to the days 3, 7, 14, and 28 after the surgery. At days 3, 7, 14, and 28, the mRNA expression of the bone morphogenetic protein-6 (BMP-6), platelet-derived growth factor-B (PDGF-B) in defect aera was analyzed by real time quantitive-polymerase blotting. HE staining was employed to reveal new bone formation in weeks 2 and 4.</p><p><b>RESULTS</b>Δ(12)-PGJ2-NC appeared opalescent white and remained relatively stable, with an average particle size of (135.2 ± 0.85) nm. The images from transmission electron microscopy showed that Δ(12)-PGJ2-NC was spherical in shape and homogeneously distributed. The encapsulation efficiency of Δ(12)-PGJ2 with the poly (lactic-co-glycolic acid) (PLGA) nanocapsules was about 92%. The in vitro release of Δ(12)-PGJ2-NC at 37 °C showed a sustained fashion and the average accumulated amount was 30%, 52%, 77%, 91%, and 98% respectively, at 0.5, 1, 2, 4 and 6 h. Compared with the animals treated with saline, after dose of 100 mg/L Δ(12)-PGJ2 and Δ(12)-PGJ2-NC apllication, the mRNA expression level of BMP-6, PDGF-B increased significantly (P < 0.05, P < 0.001). The protein expression of BMP-6, Ephrin-B2 also was up-regulated. Histomorphometry revealed that new bone formation increased at the same dose of 100 mg/L. But the unloaded nanoparticles did not have the same effect (P > 0.05).</p><p><b>CONCLUSIONS</b>A stable Δ(12)-PGJ2 loaded nanoparticle was successfully prepared. Δ(12)-PGJ2-NC may upregulate the expression of BMP-6, PDGF-B and Ephrin-B2, and promote new bone formation in bone defect area.</p>


Subject(s)
Animals , Male , Rats , Bone Morphogenetic Protein 6 , Genetics , Metabolism , Bone Regeneration , Ephrin-B2 , Genetics , Metabolism , Femur , General Surgery , Lactic Acid , Pharmacokinetics , Pharmacology , Nanocapsules , Nanoparticles , Particle Size , Polyglycolic Acid , Pharmacokinetics , Pharmacology , Prostaglandin D2 , Pharmacokinetics , Pharmacology , RNA, Messenger , Metabolism , Random Allocation , Rats, Wistar , Receptor, Platelet-Derived Growth Factor beta , Genetics , Metabolism , Time Factors , Up-Regulation
3.
Article in Chinese | WPRIM | ID: wpr-815417

ABSTRACT

OBJECTIVE@#To explore the expression profile of Ephrin-B2 in the ischemic penumbra after transient focal cerebral ischemia in rats, and to clarify the mechanism of Ephrin-B2 triggering angiogenesis.@*METHODS@#Sprague-Dawley rats were randomly divided into a normal group, a sham operation group and ischemic-reperfusion 1, 3, 7, 14, and 28 d groups. Suture-occluded method was used to establish the focal middle cerebral artery occlusion model and the ischemic brain was reperfused 2 h after the occlusion. Western blot and quantitative real-time reverse-transcription polymerase chain reaction were used to detect the dynamic expression profile of Ephrin-B2 in the penumbra cortex. Double immunofluorescence was used to speculate the location and the co-expression of Ephrin-B2 in blood vessels, neurons and astrocytes. Microvessel density was quantified by the number of CD31+ cells. Rats were subjected to neurologic functional tests by modified neurological severity scores (mNSS) before sacrifice.@*RESULTS@#Compared with the sham group, Ephrin-B2 protein and mRNA level of the penumbra cortex in the ischemic group increased 3 days (P<0.05) after the reperfusion, peaked at day 7 and 14 (P<0.01), and declined at day 28. Double immunofluorescence indicated that Ehprin-b2 was expressed in the neurons, blood vessels and astrocytes; mNSS peaked at day 7, and gradually declined at day 14. The microvessel density of penumbra cortex in the ischemic group increased 3 days (P<0.05) after the reperfusion, peaked at day 14 (P<0.01), and gradually declined at 48 h.@*CONCLUSION@#Cerebral ischemia reperfusion induces the over-expression of Ephrin-B2, with a dynamic trend, suggesting that Ehprin-b2 may improve post-stroke functional recovery by enhancing angiogenesis and neurogenesis.


Subject(s)
Animals , Rats , Astrocytes , Metabolism , Brain , Pathology , Brain Ischemia , Metabolism , Cerebral Cortex , Metabolism , Ephrin-B2 , Metabolism , Infarction, Middle Cerebral Artery , Ischemic Attack, Transient , Neurons , Metabolism , Rats, Sprague-Dawley , Reperfusion Injury , Metabolism
4.
Chinese Journal of Stomatology ; (12): 614-618, 2014.
Article in Chinese | WPRIM | ID: wpr-260766

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the mechanisms of Porphyromonas gingivalis (Pg) infection-mediated enhancement of adhesion between monocytes THP-1 and human umbilical vein endothelial cells (HUVEC) by detecting the effect of erythropoietin producing hepatomocellular receptor interacting protein B2 (Ephrin B2) and its receptors on the adhesion.</p><p><b>METHODS</b>PgATCC33277 was cultured in an anaerobic jar, and THP-1 cells were infected with various concentrations of Pg at multiplicity of infection (MOI) of 1:100 for 8 and 24 h, respectively. The expression of Ephrin B2 receptor of THP-1 cells was detected. After removal of the free Pg, THP-1 cells were cocultured with HUVEC (overexpress of EphrinB2 or not) for 24 h to detect the expression of Ephrin B2 of HUVEC cells after additional cultivation for 23 h.</p><p><b>RESULTS</b>The adhesion of THP-1 cells post infection by Pg to HUVEC was enhanced. The mRNA levels of Ephrin B2 receptors, including EphB3 (5.169±0.152, P = 0.005), EphB4 (11.040±1.195, P = 0.001), and EphA4 (4.976± 0.122, P = 0.001) expressed by THP-1, and Ephrin B2 (8.938±0.962, P = 0.008) expressed by HUVEC were significantly elevated 24 h post infection of Pg. Over expression of Ephrin B2 in HUVEC promoted the adhesion of THP-1 to HUVEC.</p><p><b>CONCLUSIONS</b>Ephrin B2 and its receptors are involved in Pg infection mediated enhancement of the adhesion of THP-1 to HUVEC cells, suggesting that Ephrin B2 participates in the development of atherosclerosis.</p>


Subject(s)
Humans , Atherosclerosis , Cell Adhesion , Cells, Cultured , Coculture Techniques , Endothelium, Vascular , Ephrin-B2 , Physiology , Epoetin Alfa , Erythropoietin , Human Umbilical Vein Endothelial Cells , Monocytes , Porphyromonas gingivalis , Virulence , Recombinant Proteins , Umbilical Veins , Cell Biology
5.
Article in Chinese | WPRIM | ID: wpr-746945

ABSTRACT

OBJECTIVE@#To investigate the expressions of EphrinB2 and VEGF in nasopharyngeal carcinoma and their relationships with clinic pathological factors.@*METHOD@#The expressions of EphrinB2 and VEGF in 136 cases of nasopharyngeal carcinomas, and 20 cases of nasopharyngitis tissues were detected by SP method immunohistochemistry.@*RESULT@#1) The positive rates of EphrinB2 and VEGF were 63.2% and 67.6% respectively in 136 cases of nasopharyngeal carcinomas. Their positive rate in nasopharyneal carcinoma tissues was significantly higher than those in nasopharyngitis tissues (P<0.01). 2) Intensity of the expression of EphrinB2 and VEGF protein was related to lymphnodes metabasis, cranial nerve palsy, basalis encroachment, clinical stage respectively (P<0.01). 3) The expression of EphrinB2 in nasopharyngeal carcinoma tissues and was positively correlated with VEGF (P< 0.01).@*CONCLUSION@#Ephrinbeta2 and VEGF may play important roles in invasiveness, metastasis and angiogenesis of nasopharyngeal carcinoma. There may be certain inter regulation mechanism between them and they are hoped to become new biologic parameters to judge the pathogenesis, development and prognosis of nasopharyngeal carcinoma and to guide the treatment.


Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Carcinoma , Ephrin-B2 , Metabolism , Immunohistochemistry , Lymphatic Metastasis , Nasopharyngeal Carcinoma , Nasopharyngeal Neoplasms , Metabolism , Pathology , Vascular Endothelial Growth Factor A , Metabolism
6.
Article in Chinese | WPRIM | ID: wpr-353036

ABSTRACT

Bidirectional Eph-Ephrin signaling as a focal point of research in cell-cell communications is critical for generation of nerves and vesssels as well as invation and metastasis of tumor cells. The roles for Ephrin-Eph bidirectional signaling in bone remodeling were important. EphrinB2 is expressed on osteoblasts and EphB4 is expressed on osteoclasts. Forward signaling through the EphB4 receptor into mesenchymal precursors promotes osteoblast differentiation, while reverse signaling through the EphrinB2 ligand into osteoclast suppresses differentiation. Signaling between the ligand EphrinB2 and the receptors EphB4 explains bidirectional signaling between osteoblasts and osteoclasts,bone absorption and remodeling, which may lay a theoretical foundation for identifying drug targeting and preventing and treating bone loss.


Subject(s)
Animals , Humans , Bone Remodeling , Physiology , Ephrin-B2 , Physiology , Osteoblasts , Cell Biology , Osteoclasts , Cell Biology , Receptor, EphB4 , Physiology , Signal Transduction , Physiology
7.
Article in Chinese | WPRIM | ID: wpr-814631

ABSTRACT

OBJECTIVE@#To explore the degree, mechanism and clinical significance of three-dimensional tumor microvascular architecture phenotype heterogeneity (3D-TMAPH) in non-small cell carcinoma (NSCLC).@*METHODS@#Twenty-one samples of solitary pulmonary nodules were collected integrally. To establish two-dimensional tumor microvascular architecture phenotype (2D-TMAP) and three-dimensional tumor microvascular architecture phenotype (3D-TMAP), five layers of each nodule were selected and embedded in paraffin. Test indices included the expressions of vascular endothelial growth factor (VEGF), proliferating cell nuclear antigen (PCNA), EphB4, ephfinB2 and microvascular density marked by anti-CD34 (CD34-MVD). The degrees of 3D-TMAPH were evaluated by the coefficient of variation and extend of heterogeneity. Spearman rank correlation analysis was used to investigate the relationships between 2D-TMAP, 3D-TMAP and clinicopathological features.@*RESULTS@#3D-TMAPH showed that 2D-TMAP heterogeneity was expressed in the tissues of NSCLC. The heterogeneities in the malignant nodules were significantly higher than those in the active inflammatory nodules and tubercular nodules. In addition, different degrees of heterogeneity of CD34-MVD and PCNA were found in NSCLC tissues. The coefficients of variation of CD34- MVD and PCNA were positively related to the degree of differentiation (all P0.05). The level of heterogeneity of various expression indexes (ephrinB2, EphB4, VEGF) in NSCLC tissues were inconsistent, but there were no significant differences in heterogeneity in NSCLC tissues with different histological types (P>0.05).@*CONCLUSION@#3D-TMAPH exists widely in the microenvironment during the genesis and development of NSCLC and has a significant impact on its biological complexity.


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Capillaries , Carcinoma, Non-Small-Cell Lung , Ephrin-B2 , Metabolism , Lung Neoplasms , Neovascularization, Pathologic , Pathology , Phenotype , Proliferating Cell Nuclear Antigen , Metabolism , Vascular Endothelial Growth Factor A , Metabolism
8.
Chinese Medical Journal ; (24): 3831-3835, 2012.
Article in English | WPRIM | ID: wpr-256633

ABSTRACT

<p><b>BACKGROUND</b>In response to the injury of the central nervous system (CNS), the astrocytes upregulate the expression of glial fibrillary acidic protein (GFAP), which largely contributes to the reactive gliosis after brain injury. The regulatory mechanism of this process is still not clear. In this study, we aimed to compare the ephrin-B2 deficient mice with the wild type ones with regard to gliosis after traumatic brain injury.</p><p><b>METHODS</b>We generated ephrin-B2 knockout mice specifically in CNS astrocytes. Twelve mice from this gene-knockout strain were randomly selected along with twelve mice from the wild type littermates. In both groups, a modified controlled cortical impact injury model was applied to create a closed traumatic brain injury. Twenty-eight days after the injury, Nissl staining and GFAP immunofluorescence staining were used to compare the brain atrophy and GFAP immunoreactivity between the two groups. All the data were analyzed by t-test for between-group comparison.</p><p><b>RESULTS</b>We successfully set up the conditional ephrin-B2 knockout mice strain, which was confirmed by genotyping and ephrin-B2/GFAP double staining. These mice developed normally without apparent abnormality in general appearance. Twenty-eight days following brain injury, histopathology revealed by immunohistochemistry showed different degrees of cerebral injuries in both groups. Compared with wild-type group, the ephrin-B2 knockout group exhibited less brain atrophy ratio for the injured hemispheres (P = 0.005) and hippocampus (P = 0.027). Also the wild-type group demonstrated greater GFAP immunoreactivity increment within hippocampal regions (P = 0.008).</p><p><b>CONCLUSIONS</b>The establishment of conditional ephrin-B2 knockout mice provides us with a new way to explore the role of ephrin-B2 in astrocytes. Our findings revealed less atrophy and GFAP immunoreactivity in the knockout mice strain after traumatic brain injury, which implied ephrin-B2 could be one of the promoters to upregulate gliosis following brain injury.</p>


Subject(s)
Animals , Mice , Atrophy , Brain , Pathology , Brain Injuries , Pathology , Ephrin-B2 , Physiology , Glial Fibrillary Acidic Protein , Gliosis , Immunohistochemistry , Mice, Inbred C57BL , Mice, Knockout , Nerve Tissue Proteins
9.
Chinese Journal of Oncology ; (12): 192-196, 2011.
Article in Chinese | WPRIM | ID: wpr-303316

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the role of the expression of ephrinB2 and EphB4 in non-small cell lung cancer (NSCLC), and their relationship with multi-slice spiral CT pulmonary perfusion imaging.</p><p><b>METHODS</b>Thirty-one nodules with pathologically proven NSCLC underwent CT perfusion scan. The perfusion parameters including blood flow (BF), blood volume (BV), peak enhancement image (PEI) were collected. The expression of ephrinB2 and EphB4 in tumor cells and interstitial vasculature were detected by immunohistochemistry. Correlation analysis and trend test were used to assess the relationship between ephrinB2/EphB4 expression and clinicopathological features, and between ephrinB2/EphB4 expression and perfusion parameters.</p><p><b>RESULTS</b>Positive expression of ephrinB2 and EphB4 in the NSCLC group was 83.9% and 71.0%, respectively, significantly higher than that in the internal control group (P < 0.01). The expression of ephrinB2 and EphB4 was consistently in tumor parenchyma but differently in tumor vessels. The expressions of ephrinB2 and EphB4 were positively correlated with lymphatic metastasis (P < 0.05). The expression of EphB4 was negatively correlated with blood flow (BF) and blood volume (BV), respectively (P < 0.05). There was a significant positive correlation between ephrinB2 expression and BF (r = 0.516, P = 0.003), and a positive correlation between ephrinB2 expression and BV (r = 0.448, P = 0.013). The expressions of ephrinB2 and EphB4 were not correlated with PEI (P > 0.05). The values of BF and BV in the high and moderate EphB4 expression groups were significantly decreased compared with that in the negative group (P < 0.01). The value of BF in the high ephrinB2 expression group was significantly increased compared with that in the moderately positive group and negative group (P < 0.01). The value of BV in the high ephrinB2 expression group was significantly increased compared with that in the negative group (P < 0.01).</p><p><b>CONCLUSION</b>The CT pulmonary perfusion imaging reflects the density difference of blood vessels with functional lumen, and such difference also depends on the quantity and quality of vasculature with functional lumen.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Blood Volume , Carcinoma, Non-Small-Cell Lung , Diagnostic Imaging , Metabolism , Pathology , Ephrin-B2 , Metabolism , Lung Neoplasms , Diagnostic Imaging , Metabolism , Pathology , Lymphatic Metastasis , Perfusion Imaging , Pulmonary Circulation , Receptor, EphB4 , Metabolism , Tomography, Spiral Computed , Methods
10.
Article in Chinese | WPRIM | ID: wpr-280094

ABSTRACT

<p><b>OBJECTIVE</b>To study the effects of ephrinB2 gene transfection on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into vascular endothelial cells.</p><p><b>METHODS</b>Wistar rat BMSCs were isolated by density gradient centrifugation and purified on the basis of their adhesion ability. The BMSCs were transfected with a lenti-virus vector encoding a constitutively active form of human ephrinB2 gene, and the cell markers including CD105, CD73, CD44, von Willebrand factor (VWF) and vascular growth factor receptor 2 (KDR) were detected using flow cytometry. The potential of ephrinB2-BMSCs for differentiation into osteoblasts and adipoblasts in vitro were tested, and the differentiation of the cells into endothelial-like cells was induced by culture in the presence of 2% fetal bovine serum and 50 ng/ml vascular endothelial growth factor.</p><p><b>RESULTS</b>EphrinB2-BMSCs were positive for the markers CD105, CD73 and CD44, and negative for the typical endothelial markers like VWF and KDR, and retained high potentials for differentiation into osteoblasts and adipoblasts in vitro after cultivation in respective media. After induced differentiation, ephrinB2-BMSCs expressed VWF and KDR and showed greater ability of differentiation into vascular endothelial cells and formation of capillary structures on matrix gel than the BMSCs without transfection.</p><p><b>CONCLUSIONS</b>EphrinB2 gene transfection efficiently promotes the differentiation of BMSCs into vascular endothelial cells. These genetically engineered cells provide valuable sources for new therapies of coronary heart disease.</p>


Subject(s)
Animals , Male , Rats , Bone Marrow Cells , Cell Biology , Metabolism , Cell Differentiation , Genetics , Physiology , Cells, Cultured , Coronary Disease , Therapeutics , Endothelial Cells , Cell Biology , Metabolism , Ephrin-B2 , Genetics , Physiology , Genetic Therapy , Methods , Mesenchymal Stem Cells , Cell Biology , Metabolism , Rats, Wistar , Transfection
11.
Chinese Medical Journal ; (24): 1815-1820, 2004.
Article in English | WPRIM | ID: wpr-257354

ABSTRACT

<p><b>BACKGROUND</b>Although there were several clinical and experimental studies discussing the pathogenesis of dural arteriovenous fistula (DAVF), the pathological process leading to intracranial DAVF so far remains unknown. In this study, we investigated the expression of vascular growth factors in order to elucidate the possible role of these factors for the development of DAVF and to study the biological activity of this uncommon lesion.</p><p><b>METHODS</b>We examined the histological features, proliferative and angiogenic capacities of the tissue specimens obtained from 6 patients who underwent surgery at our institution. Immunohistochemical staining for vascular endothelial growth factor (VEGF), its receptors Flk-1 and Flt-1, ephrin-B2, MIB-1 and proliferating cell nuclear antigen (PCNA) was performed using standard immunohistochemical techniques.</p><p><b>RESULTS</b>A positive immunostaining was found for all antibodies studied except MIB-1, whereas nuclear endothelial expression of PCNA was observed in only 3/6 cases. VEGF stained positive in all of the available specimens (6/6). Flk-1 showed a positive immunoreaction in only 2/6 cases and Flt-1 in 4/6 cases. Ephrin-B2 was expressed in the majority (5/6) of the cases.</p><p><b>CONCLUSIONS</b>These results support the hypothesis that DAVFs might be acquired dynamic vascular malformations with low biological activity. Vascular growth factors like VEGF and ephrin-B2 might play a pivotal role in the formation of DAVF.</p>


Subject(s)
Aged , Female , Humans , Male , Middle Aged , Cell Division , Central Nervous System Vascular Malformations , Metabolism , Pathology , Endothelial Cells , Cell Biology , Ephrin-B2 , Physiology , Immunohistochemistry , Ki-67 Antigen , Neovascularization, Physiologic , Proliferating Cell Nuclear Antigen , Vascular Endothelial Growth Factor A
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