Pub Date : 1995-01-01DOI: 10.3109/10623329509024651
R. McCarron, Lan Wang, D. Stanimirovic, M. Spatz
The adhesion of circulating leukocytes to vascular endothelium is a prerequisite for their emigration to extravascular tissues. The data presented here demonstrate that intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) are constitutively expressed on endothelial cell lines derived from both human brain (HBEC) and umbilical cord veins (HUVEC). Lipopolysaccharide, tumor necrosis factor-α or interferon-γ treatment of both HBEC and HUVEC cell lines up-regulated the expression of these adhesion molecules in a time- and dose-dependent manner. These same proinflammatory factors also induced the expression of E-selectin on both HBEC and HUVEC cell cultures. Endothelins (ET-1, ET-2 and ET-3) also had a similar effect on the expression of all three adhesion molecules on HBEC. However, none of the endothelins had any effects on ICAM-1, VCAM-1 or E-selectin expression by HUVEC, despite the concomitant effects of the aforementioned factors on identical cultures. These results in...
{"title":"Differential Regulation of Adhesion Molecule Expression by Human Cerebrovascular and Umbilical Vein Endothelial Cells","authors":"R. McCarron, Lan Wang, D. Stanimirovic, M. Spatz","doi":"10.3109/10623329509024651","DOIUrl":"https://doi.org/10.3109/10623329509024651","url":null,"abstract":"The adhesion of circulating leukocytes to vascular endothelium is a prerequisite for their emigration to extravascular tissues. The data presented here demonstrate that intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) are constitutively expressed on endothelial cell lines derived from both human brain (HBEC) and umbilical cord veins (HUVEC). Lipopolysaccharide, tumor necrosis factor-α or interferon-γ treatment of both HBEC and HUVEC cell lines up-regulated the expression of these adhesion molecules in a time- and dose-dependent manner. These same proinflammatory factors also induced the expression of E-selectin on both HBEC and HUVEC cell cultures. Endothelins (ET-1, ET-2 and ET-3) also had a similar effect on the expression of all three adhesion molecules on HBEC. However, none of the endothelins had any effects on ICAM-1, VCAM-1 or E-selectin expression by HUVEC, despite the concomitant effects of the aforementioned factors on identical cultures. These results in...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"46 1","pages":"339-346"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84722506","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1995-01-01DOI: 10.3109/10623329509024662
A. Burke-Gaffney, A. Keenan
In this study we have evaluated the importance of certain factors affecting the permeability of confluent monolayers of human umbilical vein endothelial cells (HUVEC) to albumin. Inclusion of serum in the assay medium and fibronectin-pretreatment of the membrane supports on which the monolayers were grown decreased basal permeability. Most importantly, the endothelial cell (EC) seeding density was demonstrated to be a crucial determinant of basal permeability. In this study, the lowest % clearance of albumin was obtained with a seeding density of 6.0 × 105 cells/cm2 (% clearance = 2.90 ± 0.08%). In contrast, lower (4.8 × 105 cells/cm2) or higher (9.91 × 105 cells/cm2) seeding densities resulted in significantly (P < 0.05) higher clearances. Permeability was increased on stimulation with tumour necrosis factor-α, interleukin-1α/β or interferon-γ and the fold increases were demonstrated to be dependent on the initial HUVEC seeding density. These results demonstrate the importance of assessing the effects of...
{"title":"Assessment of Human Endothelial Permeability In Vitro: The Importance of Cell Seeding Density","authors":"A. Burke-Gaffney, A. Keenan","doi":"10.3109/10623329509024662","DOIUrl":"https://doi.org/10.3109/10623329509024662","url":null,"abstract":"In this study we have evaluated the importance of certain factors affecting the permeability of confluent monolayers of human umbilical vein endothelial cells (HUVEC) to albumin. Inclusion of serum in the assay medium and fibronectin-pretreatment of the membrane supports on which the monolayers were grown decreased basal permeability. Most importantly, the endothelial cell (EC) seeding density was demonstrated to be a crucial determinant of basal permeability. In this study, the lowest % clearance of albumin was obtained with a seeding density of 6.0 × 105 cells/cm2 (% clearance = 2.90 ± 0.08%). In contrast, lower (4.8 × 105 cells/cm2) or higher (9.91 × 105 cells/cm2) seeding densities resulted in significantly (P < 0.05) higher clearances. Permeability was increased on stimulation with tumour necrosis factor-α, interleukin-1α/β or interferon-γ and the fold increases were demonstrated to be dependent on the initial HUVEC seeding density. These results demonstrate the importance of assessing the effects of...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"18 1","pages":"75-79"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75554007","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1995-01-01DOI: 10.3109/10623329509053388
T. Swierkosz, T. Warner, J. Vane
The release of nitric oxide (NO) by NG-hydroxy-L-arginine (L-OHArg) from endothelial cells (ECs) grown in culture was assessed by two bioassays. The first was a cascade bioassay system, in which the effluent from a column of ECs grown on beads superfused a cascade of isolated endothelium-denuded rabbit aortic strips. Exclusion of calcium from the Krebs' buffer perfusing the ECs (but not from the cascade) diminished the relaxant effects of L-OHArg. Growing the ECs in the presence of dexamethasone did not affect the relaxations of the bioassay cascade induced by L-OHArg. However, these relaxations were potentiated by superoxide dismutase (SOD) and inhibited by haemoglobin, but not inhibited by NG-nitro-L-arginine methyl ester (NO2 ArgMeE). The second bioassay was a transfer assay, in which accumulation of cGMP was measured in detector, RFL-6 fibroblast cells. The accumulation of cGMP induced by the basal release of NO from ECs was increased by L-OHArg and by SOD. However, L-OHArg, unlike SOD, did not scaven...
通过两种生物测定法评估ng -羟基- l -精氨酸(L-OHArg)对内皮细胞(ECs)释放一氧化氮(NO)的影响。第一个是级联生物测定系统，其中从珠子上生长的内皮细胞柱流出的废水与分离的内皮剥离的兔主动脉条带级联。从灌注ECs的克雷伯缓冲液中排除钙(而不是从级联中排除钙)会减弱L-OHArg的松弛作用。在地塞米松存在下生长的ECs不影响L-OHArg诱导的生物测定级联的松弛。然而，这些松弛被超氧化物歧化酶(SOD)增强，被血红蛋白抑制，但不被ng -硝基- l -精氨酸甲酯(NO2 ArgMeE)抑制。第二个生物试验是转移试验，在检测RFL-6成纤维细胞中测量cGMP的积累。L-OHArg和SOD增加了ECs基础释放NO诱导的cGMP积累。然而，与SOD不同，L-OHArg不清除…
{"title":"NG-Hydroxy-L-Arginine Releases from Endothelial Cells Nitric Oxide which Increases cGMP in RFL-6 Fibroblasts","authors":"T. Swierkosz, T. Warner, J. Vane","doi":"10.3109/10623329509053388","DOIUrl":"https://doi.org/10.3109/10623329509053388","url":null,"abstract":"The release of nitric oxide (NO) by NG-hydroxy-L-arginine (L-OHArg) from endothelial cells (ECs) grown in culture was assessed by two bioassays. The first was a cascade bioassay system, in which the effluent from a column of ECs grown on beads superfused a cascade of isolated endothelium-denuded rabbit aortic strips. Exclusion of calcium from the Krebs' buffer perfusing the ECs (but not from the cascade) diminished the relaxant effects of L-OHArg. Growing the ECs in the presence of dexamethasone did not affect the relaxations of the bioassay cascade induced by L-OHArg. However, these relaxations were potentiated by superoxide dismutase (SOD) and inhibited by haemoglobin, but not inhibited by NG-nitro-L-arginine methyl ester (NO2 ArgMeE). The second bioassay was a transfer assay, in which accumulation of cGMP was measured in detector, RFL-6 fibroblast cells. The accumulation of cGMP induced by the basal release of NO from ECs was increased by L-OHArg and by SOD. However, L-OHArg, unlike SOD, did not scaven...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"4 1","pages":"121-129"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79976937","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1995-01-01DOI: 10.3109/10623329509024654
N. Mirsky, Y. Cohen
Proliferation of blood vessels is a process necessary for the normal growth and development of tissues, and for body's repair. Several components are involved in the formation of new blood vessels: chemotactic activity which causes endothelial cells to migrate from existing capillaries into the growing zone; mitogenic activity, which includes mitoses in the endothelial cells; and the organization of the cells to form an organ the new capillary. Several factors were found which induce angiogenesis. Some of them were isolated from malignant tissues and others from normal tissues.We developed a sensitive method of endothelial cell migration towards angiogenic factors. The factors which were chosen: VEGF (Vascular Endothelial Growth Factor), and ECGF (Endothelial Cell Growth Factor). The method is based on migration of cells ABAE (Aortic Arch Derived Bovine Endothelial Cells) under agarose, towards angiogenic factors, which were seeded in wells drilled in the agarose.A dose response curve for the two compound...
{"title":"VEGF and ECGF Induce Directed Migration of Endothelial Cells: Qualitative and Quantitative Assay","authors":"N. Mirsky, Y. Cohen","doi":"10.3109/10623329509024654","DOIUrl":"https://doi.org/10.3109/10623329509024654","url":null,"abstract":"Proliferation of blood vessels is a process necessary for the normal growth and development of tissues, and for body's repair. Several components are involved in the formation of new blood vessels: chemotactic activity which causes endothelial cells to migrate from existing capillaries into the growing zone; mitogenic activity, which includes mitoses in the endothelial cells; and the organization of the cells to form an organ the new capillary. Several factors were found which induce angiogenesis. Some of them were isolated from malignant tissues and others from normal tissues.We developed a sensitive method of endothelial cell migration towards angiogenic factors. The factors which were chosen: VEGF (Vascular Endothelial Growth Factor), and ECGF (Endothelial Cell Growth Factor). The method is based on migration of cells ABAE (Aortic Arch Derived Bovine Endothelial Cells) under agarose, towards angiogenic factors, which were seeded in wells drilled in the agarose.A dose response curve for the two compound...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"7 1","pages":"13-20"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89390443","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1995-01-01DOI: 10.3109/10623329509024671
N. Hill-Kapturczak, M. Kapturczak, T. Malinski, P. Gross
A detailed overview of nitric oxide and nitric oxide synthases in the kidney is presented. Physiologically, constitutive and inducible nitric oxide synthases have been detected in basically all vascular segments of the kidney, including all large vessels and arterioles that are primarily involved in the regulation of renal hemodynamics. It was observed that nitric oxide increases renal blood flow, decreases renal vascular resistance, and exerts variable effects on glomerular filtration rate depending on the experimental conditions. In addition, macula densa generated nitric oxide appears to mediate tubuloglomerular feedback. Constitutive and inducible nitric oxide synthases have also been delineated in most renal tubular segments. The inner medullary collecting duct was shown to contain the highest amount of constitutive nitric oxide synthase as compared to other nephron segments. It appears that nitric oxide may directly enhance tubular reabsorption in the collecting duct and the proximal tubule. Pressur...
{"title":"Nitric oxide and nitric oxide synthase in the kidney: Potential roles in normal penal function and in renal dysfunction","authors":"N. Hill-Kapturczak, M. Kapturczak, T. Malinski, P. Gross","doi":"10.3109/10623329509024671","DOIUrl":"https://doi.org/10.3109/10623329509024671","url":null,"abstract":"A detailed overview of nitric oxide and nitric oxide synthases in the kidney is presented. Physiologically, constitutive and inducible nitric oxide synthases have been detected in basically all vascular segments of the kidney, including all large vessels and arterioles that are primarily involved in the regulation of renal hemodynamics. It was observed that nitric oxide increases renal blood flow, decreases renal vascular resistance, and exerts variable effects on glomerular filtration rate depending on the experimental conditions. In addition, macula densa generated nitric oxide appears to mediate tubuloglomerular feedback. Constitutive and inducible nitric oxide synthases have also been delineated in most renal tubular segments. The inner medullary collecting duct was shown to contain the highest amount of constitutive nitric oxide synthase as compared to other nephron segments. It appears that nitric oxide may directly enhance tubular reabsorption in the collecting duct and the proximal tubule. Pressur...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"48 1","pages":"253-299"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88006344","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1995-01-01DOI: 10.3109/10623329509024673
D. Ku, Si-Yen Liu, J. Dai
The present study investigated the interrelationship between nitric oxide and its intermediate degradation product, peroxynitrite, on reactivity of isolated human coronary arteries and washed human platelets. Ninety-one ring segments of human coronary arteries, with minimal intimal proliferation, were isolated from 9 patients who underwent cardiac transplantation. All vessels were pretreated with 5 μM indomethacin and precontracted with 0.5-1 μM prostaglandin F2α. Addition of 0.01-300 μM peroxynitrite produced a rapid and dose-dependent relaxation similar to that observed with isolated canine coronary arteries. The IC50 of peroxynitrite induced relaxation was 3.2 ± 0.3 μM. Superoxide dismutase (SOD, 100 u/ml) enhanced, while hemoglobin (3 μM) inhibited, the observed relaxation and changed the IC50 to 0.5 ± 0.1 μM and 63 ± 9.4 μM, respectively. In washed human platelets, addition of 1-100 μM peroxynitrite by itself did not produce any direct effect on platelets, but resulted in a dose-dependent inhibition ...
{"title":"Coronary Vascular and Antiplatelet Effects of Peroxynitrite in Human Tissues","authors":"D. Ku, Si-Yen Liu, J. Dai","doi":"10.3109/10623329509024673","DOIUrl":"https://doi.org/10.3109/10623329509024673","url":null,"abstract":"The present study investigated the interrelationship between nitric oxide and its intermediate degradation product, peroxynitrite, on reactivity of isolated human coronary arteries and washed human platelets. Ninety-one ring segments of human coronary arteries, with minimal intimal proliferation, were isolated from 9 patients who underwent cardiac transplantation. All vessels were pretreated with 5 μM indomethacin and precontracted with 0.5-1 μM prostaglandin F2α. Addition of 0.01-300 μM peroxynitrite produced a rapid and dose-dependent relaxation similar to that observed with isolated canine coronary arteries. The IC50 of peroxynitrite induced relaxation was 3.2 ± 0.3 μM. Superoxide dismutase (SOD, 100 u/ml) enhanced, while hemoglobin (3 μM) inhibited, the observed relaxation and changed the IC50 to 0.5 ± 0.1 μM and 63 ± 9.4 μM, respectively. In washed human platelets, addition of 1-100 μM peroxynitrite by itself did not produce any direct effect on platelets, but resulted in a dose-dependent inhibition ...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"33 1","pages":"309-319"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84608488","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1995-01-01DOI: 10.3109/10623329509053386
S. Reddigari, A. Kaplan
Exposure of plasma to certain negatively charged substances initiates the intrinsic coagulation pathway and bradykinin formation (Kaplan and Silverberg, 1987). The three plasma proteins required for this surface-mediated process are Factor XII (also known as Hageman factor), high molecular weight kininogen (HK) and prekallikrein. Factor XII is a monomeric beta-globulin with a MW of 80 kD and circulates in normal plasma at a concentration of 30-35 μg/ml (Cochrane and Wuepper, 1971) (Table 1). It is capable of undergoing autoactivation to the serine protease FXIIa (Silverberg, Dunn et al., 1980a; Wiggins and Cochrane, 1979) and this autoactivation is initiated by polyanionic substances such as glass, kaolin, and dextran sulfate. Known natural activators include certain mucopolysaccharides or proteoglycans (Hojima, Cochrane et al., 1984; Silverberg and Diehl, 1987), sulfatides (Shimada, Sugo et al., 1985), phospholipids (Schousboe, 1990), endotoxin (Morrison and Cochrane, 19741, and urate crystals (Ginsberg,...
血浆暴露于某些带负电荷的物质会启动内在凝血途径和缓激素的形成(Kaplan和Silverberg, 1987)。这一表面介导过程所需的三种血浆蛋白是因子XII(也称为Hageman因子)、高分子量激肽原(HK)和预钾likrein。因子XII是一种分子量为80 kD的单体β -球蛋白，在正常血浆中以30-35 μg/ml的浓度循环(Cochrane和wueper, 1971)(表1)。它能够对丝氨酸蛋白酶FXIIa进行自激活(Silverberg, Dunn等，1980a;Wiggins and Cochrane, 1979)，这种自激活是由玻璃、高岭土和硫酸葡聚糖等多阴离子物质引发的。已知的天然活化剂包括某些粘多糖或蛋白多糖(Hojima, Cochrane et al.， 1984;Silverberg和Diehl, 1987)，硫脂(Shimada, Sugo等，1985)，磷脂(Schousboe, 1990)，内毒素(Morrison和Cochrane, 19741)和尿酸盐晶体(Ginsberg，…
{"title":"Interaction of Plasma Contact Activation Factors with Vascular Endothelium","authors":"S. Reddigari, A. Kaplan","doi":"10.3109/10623329509053386","DOIUrl":"https://doi.org/10.3109/10623329509053386","url":null,"abstract":"Exposure of plasma to certain negatively charged substances initiates the intrinsic coagulation pathway and bradykinin formation (Kaplan and Silverberg, 1987). The three plasma proteins required for this surface-mediated process are Factor XII (also known as Hageman factor), high molecular weight kininogen (HK) and prekallikrein. Factor XII is a monomeric beta-globulin with a MW of 80 kD and circulates in normal plasma at a concentration of 30-35 μg/ml (Cochrane and Wuepper, 1971) (Table 1). It is capable of undergoing autoactivation to the serine protease FXIIa (Silverberg, Dunn et al., 1980a; Wiggins and Cochrane, 1979) and this autoactivation is initiated by polyanionic substances such as glass, kaolin, and dextran sulfate. Known natural activators include certain mucopolysaccharides or proteoglycans (Hojima, Cochrane et al., 1984; Silverberg and Diehl, 1987), sulfatides (Shimada, Sugo et al., 1985), phospholipids (Schousboe, 1990), endotoxin (Morrison and Cochrane, 19741, and urate crystals (Ginsberg,...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"77 1","pages":"99-111"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84362014","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1995-01-01DOI: 10.3109/10623329509024674
J. E. Freeman, W. Kuo, G. Milligan, C. Lowenstein, M. Levine, N. Flavahan
Endothelium-dependent relaxations evoked by activation of α2-adrenergic and serotonergic receptors are reduced by pertussis toxin, which irreversibly inhibits the activity of certain G-proteins. The present experiments were conducted in order to further characterize the inhibitory effect of the toxin and to directly analyze receptor: G-protein interactions in native porcine endothelial cells. Cell membranes, prepared from freshly harvested endothelial cells, were incubated with pertussis toxin in the presence of 32P-NAD and labelled proteins were separated on SDS-PAGE. Pertussis toxin catalyzed the transfer of 32P-ADP-ribose from NAD to a 40 kD protein. The toxin-catalyzed ADP-ribosylation was not significantly affected by bradykinin but was reduced by serotonin, mastoparan (a direct G-protein activator) or UK 14, 304, an α2-adrenergic agonist. Western blot analysis demonstrated that porcine endothelial cells express Giα-2 and Giα-3 protein. Immunoprecipitation of solubilized, 32P-ADP ribosylated protein ...
{"title":"Analysis of Pertussis Toxin-Sensitive Receptor: G-Protein Interactions in Native Porcine Endothelial Cells","authors":"J. E. Freeman, W. Kuo, G. Milligan, C. Lowenstein, M. Levine, N. Flavahan","doi":"10.3109/10623329509024674","DOIUrl":"https://doi.org/10.3109/10623329509024674","url":null,"abstract":"Endothelium-dependent relaxations evoked by activation of α2-adrenergic and serotonergic receptors are reduced by pertussis toxin, which irreversibly inhibits the activity of certain G-proteins. The present experiments were conducted in order to further characterize the inhibitory effect of the toxin and to directly analyze receptor: G-protein interactions in native porcine endothelial cells. Cell membranes, prepared from freshly harvested endothelial cells, were incubated with pertussis toxin in the presence of 32P-NAD and labelled proteins were separated on SDS-PAGE. Pertussis toxin catalyzed the transfer of 32P-ADP-ribose from NAD to a 40 kD protein. The toxin-catalyzed ADP-ribosylation was not significantly affected by bradykinin but was reduced by serotonin, mastoparan (a direct G-protein activator) or UK 14, 304, an α2-adrenergic agonist. Western blot analysis demonstrated that porcine endothelial cells express Giα-2 and Giα-3 protein. Immunoprecipitation of solubilized, 32P-ADP ribosylated protein ...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"11 1","pages":"321-330"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81075223","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1995-01-01DOI: 10.3109/10623329509024656
L. Juillerat-Jeanneret, P. Fioroni, P. Leuenberger
Vascular permeability is tightly controlled, in particular in the cerebral microvasculature, thus factors able to modulate permeability are of physiological importance. Interleukin-6 (IL-6) can be released by many cell types including mononuclear phagocytes, fibroblasts and endothelial cells of various origins, and is involved in a variety of cellular responses, including modulation of vascular permeability. IL-6 is synthesized as a precursor propeptide which is proteolytically processed to the active hormone, then degraded, by as yet unknown enzymatic pathways. Unstimulated brain-derived microvessels, primary cells and cell lines, all secreted bioactive IL-6. Cell lines secreted IL-6 exclusively as a 25 kD peptide. In order to understand mechanisms regulating the secretion of IL-6 in cerebral endothelium, levels of IL-6 were measured in cell culture supernatants of brain-derived endothelial cells exposed to dexamethasone, forskolin or dibutyryl cAMP and inhibitors of serine-proteases. In primary cells an...
{"title":"Modulation of Secretion of Interleukin-6 in Brain-Derived Microvascular Endothelial Cells","authors":"L. Juillerat-Jeanneret, P. Fioroni, P. Leuenberger","doi":"10.3109/10623329509024656","DOIUrl":"https://doi.org/10.3109/10623329509024656","url":null,"abstract":"Vascular permeability is tightly controlled, in particular in the cerebral microvasculature, thus factors able to modulate permeability are of physiological importance. Interleukin-6 (IL-6) can be released by many cell types including mononuclear phagocytes, fibroblasts and endothelial cells of various origins, and is involved in a variety of cellular responses, including modulation of vascular permeability. IL-6 is synthesized as a precursor propeptide which is proteolytically processed to the active hormone, then degraded, by as yet unknown enzymatic pathways. Unstimulated brain-derived microvessels, primary cells and cell lines, all secreted bioactive IL-6. Cell lines secreted IL-6 exclusively as a 25 kD peptide. In order to understand mechanisms regulating the secretion of IL-6 in cerebral endothelium, levels of IL-6 were measured in cell culture supernatants of brain-derived endothelial cells exposed to dexamethasone, forskolin or dibutyryl cAMP and inhibitors of serine-proteases. In primary cells an...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"24 1","pages":"31-37"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87490918","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1995-01-01DOI: 10.3109/10623329509024643
J. Bevan
The strategic location of the endothelium between the flowing intraluminal blood and the vascular smooth muscle cells in the blood vessel wall implies that it should play a crucial role in the blood flow regulation of vascular tone. However, the nature of many of the cellular mechanisms involved in this inter-relationship remain speculative and controversial. The author's hypothesis is that flow can initiate two responses as the result of shear stress activation of a common sensor -the characteristic response is dilation but constriction occurs at lower and can also occur at higher tone levels. At lower levels of tone, flow-contraction and -dilation interact influencing wall tone towards a balance point. Current theories of flow sensing include deformation of viscoelastic molecules attached to the lumenal surface of the endothelium, deformation of the lumenal surface of endothelial cells and flow-dependent, endogenous agonist -lumenal endothelial surface receptor interaction. There are a number of ways in...
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