Ephrin-B2 has been shown to participate in angiogenesis, but the underlying mechanisms involved remain unclear. In this study, a rat model of local cerebral ischemia was prepared by focal middle cerebral artery occlus...Ephrin-B2 has been shown to participate in angiogenesis, but the underlying mechanisms involved remain unclear. In this study, a rat model of local cerebral ischemia was prepared by focal middle cerebral artery occlusion, followed by 24-hour reperfusion. Then, ephrin-B2 protein was administered intracerebroventricularly for 3 consecutive days via a micro-osmotic pump. Western blot assay and quantitative real-time reverse transcription PCR demonstrated the expression levels of angiopoietin-1 （Ang-1） mRNA and protein in the penumbra cortex of the ephrin-B2 treated group were decreased at day 4 after reperfusion, and increased at day 28, while the expression levels of angiopoietin-2 （Ang-2） were highly up-regulated at all time points tested. Double immunofluorescent staining indicated that Ang-1 and Ang-2 were both expressed in vascular endothelial cells positive for CD31. These findings indicate that ephrin-B2 influences the expressions of Ang-1 and Ang-2 during angiogenesis following transient focal cerebral ischemia.展开更多
AIM： To evaluate the effects of angiopoietin-1 （Ang-1） on adhesion of gastric cancer cell line BGC-823 and expression of integrin β1, CD44V6, urokinase-type plasminogen activator （uPA） and matrix metalloproteina...AIM： To evaluate the effects of angiopoietin-1 （Ang-1） on adhesion of gastric cancer cell line BGC-823 and expression of integrin β1, CD44V6, urokinase-type plasminogen activator （uPA） and matrix metalloproteinase-2 （MMP-2）. METHODS： BGC-823 cells were transfected transiently with adenovirus-Ang-1 （Ad-Ang-1）. Cells transfected transiently with adenovirus-green fluorescent protein （Ad-GFP） and untransfected cells were used as a negative and blank control group, respectively. The cell adhesion rate between cell and extracellular matrix （ECM） was determined by cell adhesion assay. To investigate whether Ang-1 could reinforce gastric carcinoma metastasis, we performed migration and invasion assays in BGC-823 cells. The mRNA and protein expression of integrin β1, CD44V6, uPA and MMP-2 were detected by reverse transcription polymerase chain reaction and Western blotting, respectively. The expression of integrin β1 and CD44V6 was measured by immunohistochemistry. RESULTS： BGC-823 cells were transfected successfully. The adhesion rate increased significantly in the Ad-Ang-1 group （P 〈 0.05）. The Ad-Ang-1-transfected group had a significant increase in migration and invasion compared with that of the mock-transfected and Ad-GFP groups. The mRNA and protein expression of integrin β1, CD44V6, uPA and MMP-2 in the Ad- Ang-1 group was higher than that in the Ad-GFP and blank control groups （P 〈 0.05）. Compared with mocktransfected and Ad-GFP groups, integrin 131 and CD44V6 expression intensity greatly increased （P 〈 0.05）. CONCLUSION： Transfection of Ang-1 into human gastric cancer cell line BGC-823 can significantly increase expression of integrin β1 and CD44V6, by which cell adhesion and metastasis to the ECM are promoted.展开更多
AIM To evaluate the efficacy of peripheral blood concentrations of angiopoietins(Ang) as cirrhosis biomarkers of chronic hepatitis C(CHC).METHODS Ang1 and Ang2 serum levels were measured by enzyme-linked immunosorbent...AIM To evaluate the efficacy of peripheral blood concentrations of angiopoietins(Ang) as cirrhosis biomarkers of chronic hepatitis C(CHC).METHODS Ang1 and Ang2 serum levels were measured by enzyme-linked immunosorbent assays(ELISA) in samples from 179 cirrhotic and non-cirrhotic CHC patients, classified according to the METAVIR system.Groups were compared by non-parametric MannWhitney U test. Subsequently, the association of peripheral concentrations of angiopoietins with the stage of fibrosis was analyzed using Spearman correlation test. Finally, the accuracy, sensitivity and specificity of circulating angiopoietins for cirrhosis diagnosis were determined by the study of the respective area under the curve of receiver operator characteristics(AUC-ROC).RESULTS Peripheral blood concentrations of Ang1 and Ang2 in CHC patients were significantly related to fibrosis. While Ang1 was decreased in cirrhotic subjects compared to non-cirrhotic(P < 0.0001), Ang2 was significantly increased as CHC progressed to the end stage of liver disease(P < 0.0001). Consequently, Ang2/Ang1 ratio was notably amplified and significantly correlated with fibrosis(P < 0.0001). Interestingly, the individual performance of each angiopoietin for the diagnosis of cirrhosis reached notable AUC-ROC values(above 0.7, both), but the Ang2/Ang1 ratio was much better(AUC-ROC = 0.810) and displayed outstanding values of sensitivity(71%), specificity(84%) and accuracy(82.1%) at the optimal cut-off(10.33). Furthermore, Ang2/Ang1 ratio improved the performance of many other previously described biomarkers or scores of liver cirrhosis in CHC.CONCLUSION Ang2/Ang1 ratio might constitute a useful tool for monitoring the progression of chronic liver disease towards cirrhosis and play an important role as therapeutic target.展开更多
Introduction: Endometriosis affects up to 1 every 5 women at their reproductive age, with variable and complex symptomatology. Patients may be asymptomatic but may have pain episodes or subfertility. Its negative impa...Introduction: Endometriosis affects up to 1 every 5 women at their reproductive age, with variable and complex symptomatology. Patients may be asymptomatic but may have pain episodes or subfertility. Its negative impact is on patients’ health and quality of life. Objective: it was to investigate the serum and peritoneal fluid (PF) concentrations of Angiopoietin- 2, Interleukin-1β, and Vascular Endothelial Growth Factor, aiming to evaluate their diagnostic performance in endometriosis. Methods: Serum and peritoneal fluid samples were taken from 112 women undergoing laparoscopy for infertility, pelvic pain or adnexal masses. 61 diagnosed with endometriosis and 51 controlled. Primary outcome was to estimate serum and PF concentrations of Angio-2, IL-1β and VEGF and secondarily correlate these concentrations to disease stages thus assuming their diagnostic potential. Results: Significant differences were found between patients and control as regards serum and PF concentration of all studied markers except serum IL-1β. Serum Angio-2 and PF VEGF showed a significantly higher level in more advanced stages of endometriosis. PF VEGF showed a positively significant correlation with the stage of the disease, spearman coefficient t = 0.442 p = 0.014. PF concentrations of Angio-2 and Serum VEGF did not show significant pattern changes with stage-related levels. Diagnostic potential of serum and PF concentrations of the 3 markers were assessed by the ROC curve. Angio-2 proved an excellent diagnostic ability for endometriosis. PF and serum VEGF proved an equal diagnostic performance, whereas, PF IL-1β was the least efficient. Based on the results, we suggested preliminary serum threshold values for these markers to be used as diagnostic or follow-up landmarks with relatively acceptable sensitivity, specificity, positive and negative predictive values. Conclusion: Non-invasive predictive biomarkers for endometriosis were Serum Angio-2, IL-1β, and VEGF independently or in combination with the estimated threshold values. Serum Angio-2 merit is considered as a novel marker for endometriosis due to its diagnostic power.展开更多
AIM:To investigate angiopoietin(Ang) and vascular endothelial growth factor(VEGF) expression in rats with ulcerative colitis(UC) and colorectal cancer(CRC).METHODS:Dysplasia and cancer were investigatedin rats that re...AIM:To investigate angiopoietin(Ang) and vascular endothelial growth factor(VEGF) expression in rats with ulcerative colitis(UC) and colorectal cancer(CRC).METHODS:Dysplasia and cancer were investigatedin rats that received three cycles of 3.5% dextran sulfate sodium(DSS) in drinking water for 7 d followed by distilled water for 14 d after intraperitoneal pretreatment with 20 mg/kg 1,2-dimethylhydrazine(DMH)(CRC group).Colitis was investigated in rats that received three cycles of 3.5% DSS in drinking water for 7 d followed by distilled water for 14 d after intraperitoneal pretreatment with saline(UC group).Rats without DSS or DMH treatment served as controls.Expression of the tyrosine kinase with immunoglobulinlike and EGF-like domains(Tie)-2 and its ligands,Ang-1 and Ang-2,as well as VEGF were evaluated in the colorectum by Western blotting.RESULTS:Compared with rats in the control group,rats in the CRC and UC groups developed the symptoms of acute colitis with diarrhea,rectal bleeding,wasting,and loss of body weight(P < 0.05).In addition,the mean length of colorectum of CRC and UC rats was significantly shorter than that of control rats(8.29 ± 0.21 and 8.31 ± 0.86,respectively,vs 12.34 ± 0.12 cm; P < 0.05).Furthermore,rats in the CRC group,but not in the UC or control groups,developed multiple tumors in the colorectal region.Western blot analysis revealed that rats in the CRC and UC groups had markedly increased protein levels of Ang-1,Ang-2,Tie-2,and VEGF in the colorectum compared to rats in the control group.CONCLUSION:Increased expression of Ang-1,Ang-2,Tie-2,and VEGF in ulcerative colitis-derived colorectal cancer might lead to chronic colitis and pathologic angiogenesis in rats.展开更多
Interactions of vascular endothelial growth factor （VEGF） with receptors VEGFR1/Fltl and VEGFR2/Flk1, and those of angiopoietins （Ang-1, Ang-2） with receptor Tie2 play important roles in placental angiogenesis. Th...Interactions of vascular endothelial growth factor （VEGF） with receptors VEGFR1/Fltl and VEGFR2/Flk1, and those of angiopoietins （Ang-1, Ang-2） with receptor Tie2 play important roles in placental angiogenesis. This study investigated vascular morphology and expression of these angiogenic factors in rat placenta on the day 15, 18, 21 of gestation （D 15, D 18 and D21）. The rats were randomly assigned into 3 groups： normal group, model group [fetal growth restriction （FGR） model], and Bushen Tqi Huoxue （BYHR） recipe treatment group （BYHR group, the pregnant rats with FGR were treated with BYHR recipe）. Morphological analysis indicated that during initial villous formation, fetal nucle- ated erythrocytes （FNEs） appeared in maternal blood sinus （MBS）. Subsequently, FNEs were sur- rounded by endothelial cells to form fetal capillary （FC） and then by trophoblast cells to form villi. As pregnancy proceeded, FC density increased progressively with increasing endothelial identification staining （EIS） in normal and BYHR groups. Whereas, villous formation was suppressed, normal in- crease in FC density was impaired and EIS was weakened in model group. Quantitative PCR analysis showed that VEGF and Flkl mRNA increased over gestation in all groups, indicating that VEGF might play a pivotal role in FC growth during late gestation. VEGF mRNA was increased on D15, while de- creased on D21 in model group as compared with normal group and BYHR group. Immunohistochemi- cally, Ang-2 protein was highly expressed in FNEs, gradually disappeared as villi matured, and decreased over gestation in all groups, indicating that Ang-2 might play a pivotal role in villous formation, which was further supported by decreased Ang-2 mRNA and protein expression in model group on D 15. Ang-1 mRNA, Tie2 mRNA and Ang-1/Ang-2 ratio increased from D15 to D18 in all groups as placenta matured. Ang-1 mRNA, Tie2 mRNA and Ang-1/Ang-2 ratio were decreased on D18 in model group as compared with normal and BYHR groups, indicating delayed maturity of FGR placenta. Alterations in angiogenic factors may result in altered placental vasculature and cause placental insufficiency. BYHR recipe could balance the angiogenic factors to promote the formation and maturation of FGR placental vasculature.展开更多
Epinephrine is often used for the treatment of patients with heart failure, low cardiac output and cardiac arrest. It can acutely improve hemodynamic parameters; however, it does not seem to improve longer term clinic...Epinephrine is often used for the treatment of patients with heart failure, low cardiac output and cardiac arrest. It can acutely improve hemodynamic parameters; however, it does not seem to improve longer term clinical outcomes. Therefore, we hypothesized that epinephrine may induce unfavorable changes in gene expression of cardiomyocyte. Thus, we investigated effects of epinephrine exposure on the mediation or modulation of gene expression of cultured cardiomyocytes at a genome-wide scale. Our investigation revealed that exposure of cardiomyocytes to epinephrine in an in vitro environment can up-regulate the expression ofangiopoietin-2 gene （~ 2.1 times）, and down-regulate the gene expression of neuregulin 1 （-3.7 times）, plasminogen activator inhibitor-1 （-2.4 times） and SPARC-related modular calcium-binding protein-2 （-4.5 times）. These changes suggest that epinephrine exposure may induce inhibition of angiogenesis-related gene expressions in cultured rat cardiomyocytes. The precise clinical significance of these changes in gene expression, which was induced by epinephrine exposure, warrants further experimental and clinical investigations.展开更多
文摘Ephrin-B2 has been shown to participate in angiogenesis, but the underlying mechanisms involved remain unclear. In this study, a rat model of local cerebral ischemia was prepared by focal middle cerebral artery occlusion, followed by 24-hour reperfusion. Then, ephrin-B2 protein was administered intracerebroventricularly for 3 consecutive days via a micro-osmotic pump. Western blot assay and quantitative real-time reverse transcription PCR demonstrated the expression levels of angiopoietin-1 （Ang-1） mRNA and protein in the penumbra cortex of the ephrin-B2 treated group were decreased at day 4 after reperfusion, and increased at day 28, while the expression levels of angiopoietin-2 （Ang-2） were highly up-regulated at all time points tested. Double immunofluorescent staining indicated that Ang-1 and Ang-2 were both expressed in vascular endothelial cells positive for CD31. These findings indicate that ephrin-B2 influences the expressions of Ang-1 and Ang-2 during angiogenesis following transient focal cerebral ischemia.
文摘AIM： To evaluate the effects of angiopoietin-1 （Ang-1） on adhesion of gastric cancer cell line BGC-823 and expression of integrin β1, CD44V6, urokinase-type plasminogen activator （uPA） and matrix metalloproteinase-2 （MMP-2）. METHODS： BGC-823 cells were transfected transiently with adenovirus-Ang-1 （Ad-Ang-1）. Cells transfected transiently with adenovirus-green fluorescent protein （Ad-GFP） and untransfected cells were used as a negative and blank control group, respectively. The cell adhesion rate between cell and extracellular matrix （ECM） was determined by cell adhesion assay. To investigate whether Ang-1 could reinforce gastric carcinoma metastasis, we performed migration and invasion assays in BGC-823 cells. The mRNA and protein expression of integrin β1, CD44V6, uPA and MMP-2 were detected by reverse transcription polymerase chain reaction and Western blotting, respectively. The expression of integrin β1 and CD44V6 was measured by immunohistochemistry. RESULTS： BGC-823 cells were transfected successfully. The adhesion rate increased significantly in the Ad-Ang-1 group （P 〈 0.05）. The Ad-Ang-1-transfected group had a significant increase in migration and invasion compared with that of the mock-transfected and Ad-GFP groups. The mRNA and protein expression of integrin β1, CD44V6, uPA and MMP-2 in the Ad- Ang-1 group was higher than that in the Ad-GFP and blank control groups （P 〈 0.05）. Compared with mocktransfected and Ad-GFP groups, integrin 131 and CD44V6 expression intensity greatly increased （P 〈 0.05）. CONCLUSION： Transfection of Ang-1 into human gastric cancer cell line BGC-823 can significantly increase expression of integrin β1 and CD44V6, by which cell adhesion and metastasis to the ECM are promoted.
基金Supported by the Ministerio de Ciencia e Innovación(SAF:2010/21805,partially)CIBERehd(Instituto de Salud CarlosⅢ,Madrid)+1 种基金Fundación Mutua Madrile■a(to Moreno-Otero R)a grant from Asociación Espa■ola Contra el Cáncer(AIO 2010,AECC,to Sanz-Cameno P)
文摘AIM To evaluate the efficacy of peripheral blood concentrations of angiopoietins(Ang) as cirrhosis biomarkers of chronic hepatitis C(CHC).METHODS Ang1 and Ang2 serum levels were measured by enzyme-linked immunosorbent assays(ELISA) in samples from 179 cirrhotic and non-cirrhotic CHC patients, classified according to the METAVIR system.Groups were compared by non-parametric MannWhitney U test. Subsequently, the association of peripheral concentrations of angiopoietins with the stage of fibrosis was analyzed using Spearman correlation test. Finally, the accuracy, sensitivity and specificity of circulating angiopoietins for cirrhosis diagnosis were determined by the study of the respective area under the curve of receiver operator characteristics(AUC-ROC).RESULTS Peripheral blood concentrations of Ang1 and Ang2 in CHC patients were significantly related to fibrosis. While Ang1 was decreased in cirrhotic subjects compared to non-cirrhotic(P < 0.0001), Ang2 was significantly increased as CHC progressed to the end stage of liver disease(P < 0.0001). Consequently, Ang2/Ang1 ratio was notably amplified and significantly correlated with fibrosis(P < 0.0001). Interestingly, the individual performance of each angiopoietin for the diagnosis of cirrhosis reached notable AUC-ROC values(above 0.7, both), but the Ang2/Ang1 ratio was much better(AUC-ROC = 0.810) and displayed outstanding values of sensitivity(71%), specificity(84%) and accuracy(82.1%) at the optimal cut-off(10.33). Furthermore, Ang2/Ang1 ratio improved the performance of many other previously described biomarkers or scores of liver cirrhosis in CHC.CONCLUSION Ang2/Ang1 ratio might constitute a useful tool for monitoring the progression of chronic liver disease towards cirrhosis and play an important role as therapeutic target.
文摘Introduction: Endometriosis affects up to 1 every 5 women at their reproductive age, with variable and complex symptomatology. Patients may be asymptomatic but may have pain episodes or subfertility. Its negative impact is on patients’ health and quality of life. Objective: it was to investigate the serum and peritoneal fluid (PF) concentrations of Angiopoietin- 2, Interleukin-1β, and Vascular Endothelial Growth Factor, aiming to evaluate their diagnostic performance in endometriosis. Methods: Serum and peritoneal fluid samples were taken from 112 women undergoing laparoscopy for infertility, pelvic pain or adnexal masses. 61 diagnosed with endometriosis and 51 controlled. Primary outcome was to estimate serum and PF concentrations of Angio-2, IL-1β and VEGF and secondarily correlate these concentrations to disease stages thus assuming their diagnostic potential. Results: Significant differences were found between patients and control as regards serum and PF concentration of all studied markers except serum IL-1β. Serum Angio-2 and PF VEGF showed a significantly higher level in more advanced stages of endometriosis. PF VEGF showed a positively significant correlation with the stage of the disease, spearman coefficient t = 0.442 p = 0.014. PF concentrations of Angio-2 and Serum VEGF did not show significant pattern changes with stage-related levels. Diagnostic potential of serum and PF concentrations of the 3 markers were assessed by the ROC curve. Angio-2 proved an excellent diagnostic ability for endometriosis. PF and serum VEGF proved an equal diagnostic performance, whereas, PF IL-1β was the least efficient. Based on the results, we suggested preliminary serum threshold values for these markers to be used as diagnostic or follow-up landmarks with relatively acceptable sensitivity, specificity, positive and negative predictive values. Conclusion: Non-invasive predictive biomarkers for endometriosis were Serum Angio-2, IL-1β, and VEGF independently or in combination with the estimated threshold values. Serum Angio-2 merit is considered as a novel marker for endometriosis due to its diagnostic power.
基金Supported by Science and Technology Project of Liaoning Province,No.2013225303the Science and Technology Project of Shenyang City,No.F13-316-1-40
文摘AIM:To investigate angiopoietin(Ang) and vascular endothelial growth factor(VEGF) expression in rats with ulcerative colitis(UC) and colorectal cancer(CRC).METHODS:Dysplasia and cancer were investigatedin rats that received three cycles of 3.5% dextran sulfate sodium(DSS) in drinking water for 7 d followed by distilled water for 14 d after intraperitoneal pretreatment with 20 mg/kg 1,2-dimethylhydrazine(DMH)(CRC group).Colitis was investigated in rats that received three cycles of 3.5% DSS in drinking water for 7 d followed by distilled water for 14 d after intraperitoneal pretreatment with saline(UC group).Rats without DSS or DMH treatment served as controls.Expression of the tyrosine kinase with immunoglobulinlike and EGF-like domains(Tie)-2 and its ligands,Ang-1 and Ang-2,as well as VEGF were evaluated in the colorectum by Western blotting.RESULTS:Compared with rats in the control group,rats in the CRC and UC groups developed the symptoms of acute colitis with diarrhea,rectal bleeding,wasting,and loss of body weight(P < 0.05).In addition,the mean length of colorectum of CRC and UC rats was significantly shorter than that of control rats(8.29 ± 0.21 and 8.31 ± 0.86,respectively,vs 12.34 ± 0.12 cm; P < 0.05).Furthermore,rats in the CRC group,but not in the UC or control groups,developed multiple tumors in the colorectal region.Western blot analysis revealed that rats in the CRC and UC groups had markedly increased protein levels of Ang-1,Ang-2,Tie-2,and VEGF in the colorectum compared to rats in the control group.CONCLUSION:Increased expression of Ang-1,Ang-2,Tie-2,and VEGF in ulcerative colitis-derived colorectal cancer might lead to chronic colitis and pathologic angiogenesis in rats.
基金supported by the National Natural Science Foundation of China(No.30973833)
文摘Interactions of vascular endothelial growth factor （VEGF） with receptors VEGFR1/Fltl and VEGFR2/Flk1, and those of angiopoietins （Ang-1, Ang-2） with receptor Tie2 play important roles in placental angiogenesis. This study investigated vascular morphology and expression of these angiogenic factors in rat placenta on the day 15, 18, 21 of gestation （D 15, D 18 and D21）. The rats were randomly assigned into 3 groups： normal group, model group [fetal growth restriction （FGR） model], and Bushen Tqi Huoxue （BYHR） recipe treatment group （BYHR group, the pregnant rats with FGR were treated with BYHR recipe）. Morphological analysis indicated that during initial villous formation, fetal nucle- ated erythrocytes （FNEs） appeared in maternal blood sinus （MBS）. Subsequently, FNEs were sur- rounded by endothelial cells to form fetal capillary （FC） and then by trophoblast cells to form villi. As pregnancy proceeded, FC density increased progressively with increasing endothelial identification staining （EIS） in normal and BYHR groups. Whereas, villous formation was suppressed, normal in- crease in FC density was impaired and EIS was weakened in model group. Quantitative PCR analysis showed that VEGF and Flkl mRNA increased over gestation in all groups, indicating that VEGF might play a pivotal role in FC growth during late gestation. VEGF mRNA was increased on D15, while de- creased on D21 in model group as compared with normal group and BYHR group. Immunohistochemi- cally, Ang-2 protein was highly expressed in FNEs, gradually disappeared as villi matured, and decreased over gestation in all groups, indicating that Ang-2 might play a pivotal role in villous formation, which was further supported by decreased Ang-2 mRNA and protein expression in model group on D 15. Ang-1 mRNA, Tie2 mRNA and Ang-1/Ang-2 ratio increased from D15 to D18 in all groups as placenta matured. Ang-1 mRNA, Tie2 mRNA and Ang-1/Ang-2 ratio were decreased on D18 in model group as compared with normal and BYHR groups, indicating delayed maturity of FGR placenta. Alterations in angiogenic factors may result in altered placental vasculature and cause placental insufficiency. BYHR recipe could balance the angiogenic factors to promote the formation and maturation of FGR placental vasculature.
基金supported by internal funding from the Department of Anesthesiology and Perioperative Medicine
文摘Epinephrine is often used for the treatment of patients with heart failure, low cardiac output and cardiac arrest. It can acutely improve hemodynamic parameters; however, it does not seem to improve longer term clinical outcomes. Therefore, we hypothesized that epinephrine may induce unfavorable changes in gene expression of cardiomyocyte. Thus, we investigated effects of epinephrine exposure on the mediation or modulation of gene expression of cultured cardiomyocytes at a genome-wide scale. Our investigation revealed that exposure of cardiomyocytes to epinephrine in an in vitro environment can up-regulate the expression ofangiopoietin-2 gene （~ 2.1 times）, and down-regulate the gene expression of neuregulin 1 （-3.7 times）, plasminogen activator inhibitor-1 （-2.4 times） and SPARC-related modular calcium-binding protein-2 （-4.5 times）. These changes suggest that epinephrine exposure may induce inhibition of angiogenesis-related gene expressions in cultured rat cardiomyocytes. The precise clinical significance of these changes in gene expression, which was induced by epinephrine exposure, warrants further experimental and clinical investigations.
