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PubMed1984

Quantitation of Na/K ATPase pump sites in the rabbit corneal endothelium.

In these experiments, the binding of 3H . ouabain, a specific inhibitor of Na/K ATPase, was used to quantitate the density of Na/K ATPase pump sites in the rabbit corneal endothelium. The uptake of ouabain by the corneal endothelium shows two components: one that saturates at a ouabain concentration near 2 X 10(-7) M (specific binding), and one component that increases linearly with increasing glycoside concentration (nonspecific uptake). The nonspecific uptake can be accounted for by that ouabain equilibrating with the extracellular space, which, estimated by inulin space, amounts to 13.0 nl/mm2 of endothelium. The saturable component of endothelial ouabain uptake is displaced by K+ ions, which is consistent with this fraction being bound to Na/K ATPase. Maximal endothelial ouabain binding was measured as 20.7 fmoles/mm2 of endothelium, which corresponds to 3.0 X 10(6) pump sites per cell. The density of Na/K ATPase pump sites in the rabbit corneal endothelium is comparable to densities reported for several transporting epithelia. These data are consistent with the known function of the endothelium in corneal deturgescense and corroborate the importance of Na/K ATPase in endothelial fluid transport.

Animals↗

PubMed1993

Nitric oxide-mediated, endothelium-dependent vasodilation is selectively attenuated in the postischemic extremity.

BACKGROUND: Attenuation of endothelium-dependent relaxing factor (EDRF) release may contribute to adverse sequelae commonly seen after reperfusion of an acutely ischemic extremity. The purpose of this study was to identify the compound responsible for the EDRF activity in the extremity and to evaluate its modulation by ischemia and reperfusion. METHODS: Isolated rat hindlimbs were perfused at constant pressure with an albumin-enriched crystalloid buffer. Increasing log dose infusions of acetylcholine and nitroprusside were used to measure endothelium-dependent (EDRF-mediated) and endothelium-independent vasoreactivity, respectively. RESULTS: Graded reductions in total vascular resistance were seen in response to both agonists in the control group (n = 11). In the postischemic group (n = 7), 60 minutes of normothermic ischemia and 10 minutes of reperfusion reduced endothelium-dependent vasodilation to acetylcholine by approximately 40% (p < 0.01). Endothelium-independent vasodilation to nitroprusside was unaffected. NG-monomethyl-L-arginine (L-NMMA), a competitive inhibitor of nitric oxide release, attenuated vasodilation to acetylcholine (p < 0.01) but not nitroprusside in both control and postischemic groups. CONCLUSIONS: Endothelium-dependent vasodilation in the rat hindlimb, mediated by nitric oxide, was selectively impaired by injury from ischemia and reperfusion. Strategies designed to minimize postischemic attenuation of nitric oxide release may be beneficial in the management of acute limb ischemia.

Acetylcholine↗

PubMed1997

Vascular reactivity to endothelium-derived relaxing factor in human umbilical artery at term pregnancy.

It has been reported that human umbilical artery (HUA) at term pregnancy released endothelium-derived relaxing factor (EDRF), using a superfusion bioassay system. However, other reports showed that endothelium-dependent relaxation was not observed in isometric tension studies using HUA ring with intact endothelium. Thus, we intended to clarify whether vascular smooth muscle of HUA at term is sensitive to EDRF. HUA was obtained after normal vaginal delivery or cesarean section at term. Isometric tension studies were performed in normal Krebs solution, using HUA rings or strips, which were prepared in calcium-free Krebs solution. Sodium nitroprusside (SNP), a nitric oxide (NO) donor drug, relaxed HUA rings precontracted with 0.1 microM 5-hydroxytryptamine (5HT) in a dose-dependent manner (1 nM-10 microM). Histamine, substance P, carbachol, or the calcium ionophore A23187, which are considered to be EDRF-releasing agents, did not relax the HUA rings. By immunohistochemical study, it was confirmed that endothelial cells were present in the luminal surface of the HUA rings after the isometric tension recording. In a co-axial bioassay system involving HUA strips denuded of endothelium and rabbit aorta with intact endothelium, HUA strips precontracted with 0.1 microM 5HT were relaxed in response to 1 microM SNP but not 1 microM carbachol, which released EDRF from the endothelium of rabbit aorta. These findings suggest that HUA at term is sensitive to NO but not EDRF.

Animals↗

PubMed1996

Endothelium-dependent effect of perindopril and enalaprilat in rat thoracic aorta.

AIM: To study the effect of the angiotensin-converting enzyme (ACE) inhibitors perindopril (Per) and enalaprilat (Ena) on the reactivity of the endothelium in normal rats. METHODS: Male rats were treated intragastrically with Per (2 mg.kg-1.d-1) or placebo (n = 18) for 6 wk. Aorta was isolated for experiment. Another set of isolated aortic rings with and without endothelium were incubated with Ena (0.1 mumol.L-1) for 30 min. Responses to acetylcholine, serotonin, phenylephrine, sodium nitroprusside (SN), and nitroglycerin (Nit) were observed. RESULTS: Endothelium-dependent relaxation to acetylcholine was augmented in aortic rings from rats treated with Per in comparison with control. The IC50 value (95% confidence limits) decreased from 3.8 (0.56-26.1) mumol.L-1 (control group) to 0.98 (0.28-3.41) mumol.L-1 (Per-treated group). The maximal relaxation was augmented from 62 +/- 9% to 78 +/- 10% (P < 0.01). However, the responses to the endothelium-independent vasodilators, SN and Nit, were similar. Serotonin- and phenylephrine-induced contractions were decreased, which were influenced by basal release of endothelium-derived relaxing factor (EDRF). EC50 values was 6.1 (2.6-14.4) nmol.L-1 vs 8.3 (3.6-18.8) nmol.L-1 in comparison with control group and Per-treated group. The maximal contraction was decreased from 2.42 +/- 0.29 g (control group) to 1.96 +/- 0.25 g (treated group) (P < 0.01). Similar results were found in incubation with Ena. CONCLUSION: Ena and Per enhanced the basic release of EDRF from vascular endothelium.

Angiotensin-Converting Enzyme Inhibitors↗

PubMed2003

Endothelium dysfunction in LDL receptor knockout mice: a role for H2O2.

1. In this study, the role of endogenous H(2)O(2) as an endothelium-dependent relaxant factor was characterised in aortas from C57BL/6J and LDL receptor-deficient mice (LDLR(-/-)). 2. Aortic rings from LDLR(-/-) mice showed impaired endothelium-dependent relaxation to acetylcholine (ACh; 0.001-100 micro M) and to the Ca(2+) ionophore A23187 (0.001-3 micro M) compared with aortic rings from control mice. Endothelium-independent relaxation produced by the NO donor, 3-morpholino-sydnonimine (SIN-1) was not different between strains. 3. Pretreatment of vessels with L-NNA (100 micro M) or L-NNA (100 micro M) plus L-NAME (300 micro M) plus haemoglobin (10 micro M) markedly decreased, but did not abolish the relaxation to ACh in control mice. In the aortas from LDLR(-/-) mice treated with L-NNA (100 micro M), ACh induced a contractile effect. Catalase (800 and 2400 U ml(-1)) shifted to the right the endothelium-dependent relaxation to ACh in aortas from control but not from LDLR(-/-) mice. Aminotriazole (50 mM), which inhibits catalase, abolished its effect on control mice. Treatment of vessels with L-NNA and catalase abolished vasorelaxation induced by ACh. Indomethacin (10 micro M) did not modify the concentration-response curve to ACh. Superoxide dismutase (300 U ml(-1)) did not change ACh-induced relaxation in both strains. 4. Exogenous H(2)O(2) produced a concentration-dependent relaxation in endothelium-denuded aortic rings, which was not different between strains. 5. It is concluded that H(2)O(2) greatly contributes to relaxation to ACh in aorta from control mice. Endothelial-dependent relaxation to ACh is impaired in LDLR(-/-) mice. Reduced biosynthesis or increased inactivation of H(2)O(2) is the possible mechanism responsible for endothelial dysfunction in aortas of atherosclerosis-susceptible LDLR(-/-) mice.

Acetylcholine↗

PubMed2001

Interactions of human prostatic epithelial cells with bone marrow endothelium: binding and invasion.

Prostate cancer shows a propensity to form secondary tumours within the bone marrow. Such tumours are the major cause of mortality in this disease. We have developed an in vitro system to study the binding of prostate epithelial cells to bone marrow endothelium (BME) and stroma (BMS). The metastatic prostate cancer cell line, PC3 (derived from a bone metastasis), was seeded onto confluent layers of BME and its binding characteristics compared to human umbilical vein endothelial cells (HUVEC), lung endothelium (Hs888Lu) and BMS. The PC3 cell line showed significantly increased binding to BME (P< 0.05) compared to endothelium derived from HUVEC and lung or BMS with maximal binding occurring at 1 h. Following pre-incubation with a beta1 integrin antibody PC3 binding to BME was inhibited by 64% (P< 0.001). Antibodies directed against the integrins beta4, alpha2, alpha4, alpha5 and the cellular adhesion molecules P-selectin, CD31, VCAM-1 and sialy Lewis X showed no effect on blocking PC3 binding. Primary prostatic epithelial cells from both malignant (n = 11) and non-malignant tissue (n = 11) also demonstrated equivalent levels of increased adhesion to BME and BMS compared to HUVEC, peaking at 24 h. Further studies examined the invasive ability of prostate epithelial cells in response to bone marrow endothelium using Matrigel invasion chamber assays. In contrast to the previous results, malignant cells showed an increase (1000 fold) in invasive ability, whilst non-malignant prostate epithelia did not respond. We have shown that both malignant and non-malignant prostate epithelial cells can bind at equivalent levels and preferentially to primary human bone marrow endothelium in comparison to controls. However, only malignant prostate epithelia show increased invasive ability in response to BME.

Adenocarcinoma↗

PubMed2003

Pivotal role of Cu,Zn-superoxide dismutase in endothelium-dependent hyperpolarization.

The endothelium plays an important role in maintaining vascular homeostasis by synthesizing and releasing several vasodilating factors, including prostacyclin, NO, and endothelium-derived hyperpolarizing factor (EDHF). We have recently identified that endothelium-derived H2O2 is an EDHF in mesenteric arteries of mice and humans and in porcine coronary microvessels. However, the mechanism for the endothelial production of H2O2 as an EDHF remains to be elucidated. In this study, we tested our hypothesis that Cu,Zn-superoxide dismutase (Cu,Zn-SOD) plays a pivotal role in endothelium-dependent hyperpolarization, using control and Cu,Zn-SOD-/- mice. In mesenteric arteries, EDHF-mediated relaxations and hyperpolarizations were significantly reduced in Cu,Zn-SOD-/- mice with no inhibitory effect of catalase, while endothelium-independent relaxations and hyperpolarizations were preserved. Endothelial H2O2 production also was significantly reduced in Cu,Zn-SOD-/- mice. In Langendorff isolated heart, bradykinin-induced increase in coronary flow was significantly reduced in Cu,Zn-SOD-/- mice, again with no inhibitory effect of catalase. The exogenous SOD mimetic tempol significantly improved EDHF-mediated relaxations and hyperpolarizations and coronary flow response in Cu,Zn-SOD-/- mice. These results prove the novel concept that endothelial Cu,Zn-SOD plays an important role as an "EDHF synthase" in mice, in addition to its classical role to scavenge superoxide anions.

Animals↗

PubMed1996

Inhibition of plasma-mediated adherence of sickle erythrocytes to microvascular endothelium by conformationally constrained RGD-containing peptides.

Adherence of sickle erythrocytes to vascular endothelium likely initiates or participates in microvascular occlusion, leading to ischemic tissue and organ damage characteristic of sickle-cell pain episodes. In vitro, sickle-cell adherence to endothelium involves adhesive plasma proteins and integrin and nonintegrin receptors on sickle cells and endothelial cells. The involvement of arginine-glycine-aspartic acid (RGD) sequences in adhesive plasma proteins and integrin receptors suggests that RGD-containing peptides may inhibit sickle-cell/endothelial-cell adherence. In the present study, inhibition of plasma-mediated sickle-erythrocyte adherence to endothelium using conformationally constrained RGD-containing peptides was quantified in vitro under continuous flow at a shear stress of 1.0 dyn/cm2. Two conformationally constrained RGD peptides were investigated: 6Z (which has high affinity for alpha5beta1, alpha(V)beta3, and alpha(IIIb)beta3 integrin receptors), and TP9201 (which preferentially binds to alpha(IIb)beta3). Peptide 6Z at 50 microM inhibited plasma-mediated sickle-cell adherence to microvascular endothelium 70% when incubated with sickle red cells, and 63% when incubated with endothelium. Under similar conditions, peptide TP9201 inhibited plasma-mediated sickle-cell adherence up to 85% at concentrations from 250 to 500 microM TP9201. The inhibition of plasma-mediated adherence by conformationally constrained RGD peptides, but not by linear or circular constructs, suggests that the tertiary structure of the peptide containing the binding sequence is important. Inhibition of plasma-mediated sickle-cell adhesion with these peptides in vitro suggests that such conformationally constrained RGD peptides could provide therapeutic interventions in the course of the disease by inhibiting receptor-ligand interactions.

Anemia, Sickle Cell↗

PubMed2000

Endothelium-derived factors as paracrine mediators of prostate cancer progression.

BACKGROUND: Vascular endothelium represents a complex network of cells producing a large number of active substrates affecting physiologic, metabolic, and immunologic properties of the whole organism, as well as particular organs or tissues. The potential influence of endothelium-derived paracrine factors on prostate cancer progression has only begun to be examined. METHODS: This review summarizes recent literature on endothelium-derived factors, including vasoactive agents, peptide growth factors, cytokines, and colony-stimulating factors, involved in the development and progression of prostate cancer. RESULTS: Endothelial cells produce an array of active substrates, many of which have been shown to influence prostate cancer growth. Available data demonstrate the positive impact of such molecules as endothelin-1, basic FGF, TGF-beta, IL-6, and IL-8 on prostate cancer progression. Many other endothelium-derived factors NO, IGF, PDGF, IL-1, G-CSF, and GM-CSF (Nitric Oxide, Insulin-Like Growth Factor, Platelet-Derived Growth Factor, Interleukin-1, Granulocyte Colony Stimulating Factor, and Granulocyte-Macrophage Colony Stimulating Factor) are, at best, implicated in prostate cancer growth, and in most cases support cancer progression. CONCLUSIONS: A better understanding of endothelium-derived factors, as paracrine mediators of prostate carcinogenesis and progression, should aid in the development of novel therapeutic strategies.

Cell Division↗

PubMed2000

Endothelium-dependent vasodilatation in patients with familial amyloidotic polyneuropathy.

We examined endothelium-dependent vasodilatation in 15 familial amyloidotic polyneuropathy (FAP) amyloidogenic transthyretin (ATTR) Valine30Methionine (Val30Met) patients and 12 healthy volunteers. Using ultrasonography, we measured the radial artery diameters under both baseline and hyperemic conditions. Endothelium-dependent vasodilatation was expressed as a percent increase in the diameters of the radial artery after induced hyperemia. Endothelium-dependent vasodilatation tended to decrease in the patients, compared with healthy volunteers. Responses were not elicited at all in patients with disease of more than 9 years' duration. Linear negative correlation was observed between endothelium-dependent vasodilatation and disease duration (P < 0.01). Correlation between endothelium-dependent vasodilatation and degree of autonomic dysfunction was significant (P = 0.0524) and for age was close to significance (P = 0.051). These results suggest that the peripheral vasomotor dysfunction in FAP patients may predominantly depend on the amount of amyloid deposition around the vessels through the course of illness.

Adult↗

PubMed2002

Sickle erythrocyte adherence to endothelium at low shear: role of shear stress in propagation of vaso-occlusion.

Under venular flow conditions, sickle cell adherence to endothelium is mediated by cell adhesion molecules and adhesive proteins associated with inflammation, coagulation, and endothelial perturbation. Periodic and reduced blood flow are observed in sickle microcirculation during hematologic steady state, suggesting that blood flow is compromised in sickle microcirculation. We tested the hypothesis that low blood flow enhances adherence by quantifying sickle cell adhesion to endothelium under venular flow (1.0 dyne/cm(2) shear stress) and low flow (0.1 dyne/cm(2) shear stress), with and without addition of adhesion promoting agonists. Under low flow, sickle cell adherence to endothelium increases with contact time in the absence of endothelial activation or adhesive protein addition. In contrast, at venular shear stress, sickle cell adherence only occurs following endothelial activation with TNF-alpha or addition of thrombospondin. Analysis of these data with a mathematical model reveals that at low flow adherence is "transport-controlled," meaning that contact time between sickle cells and endothelium is a more important determinant of adherence than high-affinity receptor-ligand interactions. Low-affinity interactions are sufficient for adhesion at low flow. In contrast, at venular flow (1 dyne/cm(2) shear stress) adherence is "affinity-controlled," meaning that adherence requires induction of specific high-affinity receptor-ligand interactions. These findings demonstrate that in addition to activating factors and adherence proteins, microvascular shear stress is an important determinant of sickle cell adhesion to endothelium. This suggests that in vivo, erythrostasis is an important determinant of adhesion that can act either independently or concurrently with ongoing acute events to induce adhesive interactions and vaso-occlusion.

Anemia, Sickle Cell↗

PubMed1990

Enhancement of sickle erythrocyte adherence to endothelium by autologous platelets.

The increased adhesiveness of sickle erythrocytes (SS RBC) to endothelial cells has been confirmed in a static system utilizing fresh umbilical vein endothelium. Adherence of SS RBC to the endothelium was as great in the presence of calcium-containing buffer as when incubated in plasma. SS RBC suspended in autologous platelet-rich plasma adhered to a greater extent than when suspended in autologous platelet poor plasma. Prostacyclin, thromboxane B2, and an inhibitor of collagen- and epinephrine-induced platelet aggregation (B13.177) did not affect SS RBC adherence to endothelium. Aspirin in a concentration of 5 micrograms/ml slightly decreased SS RBC adherence to endothelium in the presence of platelets. Platelets may play a significant role in the increased adhesiveness of SS RBC to endothelium. To the extent that increased SS RBC adhesiveness contributes to the genesis of painful crises and to the extent platelets augment this adhesiveness, agents affecting platelet function may prove useful in preventing painful crises.

Anemia, Sickle Cell↗

PubMed1988

Interaction between lymphocytes and cultured high endothelial cells: an in vitro model of lymphocyte migration across high endothelial venule endothelium.

The interaction between lymphocytes and cultured high endothelial venule endothelium has been studied using light and electron microscopy. High endothelial cells (HEC) in monolayer culture bound lymphocytes 15-200-fold more efficiently than aortic endothelium, aortic fibroblasts or serum-coated glass. Light microscopy of lymphocytes bound to HEC showed two populations. Type I lymphocytes were phase-light, round and revealed no intracellular detail. Type II lymphocytes were phase-dark and flattened. The nucleus and cytoplasm of type II cells were clearly visible under high power light microscopy. The relative positions of these two populations were determined by electron microscopy. Type I lymphocytes were attached to the surface of HEC and type II lymphocytes were flattened underneath. The transition from type I to type II was accompanied by a loss of surface microvilli and a redistribution of intracellular organelles. This suggested that lymphocytes actively migrated across the HEC layer in this assay. The relationship between migrated lymphocytes and total adherent cells was determined by analysis of surface phenotype. Lymphocytes did not adhere randomly from the cell population plated. After 60 min there was an enrichment for B over T lymphocytes and the adherent T cell population was itself enriched for CD8+ cells over CD4+ cells. Migrated cells were, however, a random subpopulation of lymphocytes which adhered to HEC. This is clear evidence that migration was preceded by specific binding to HEC. Lymphocyte adhesion was independent of viable HEC showing that it was a passive event on the part of the endothelium. Lymphocyte migration was, however, completely dependent on viable high endothelium. We conclude that cultured HEC provide a biologically relevant, 3-dimensional matrix which supports the specific adhesion of lymphocytes and actively promotes lymphocyte migration. These observations suggest to us that cultured high endothelium provides a novel in vitro model for the study of lymphocyte migration into lymph nodes from the blood.

Animals↗

PubMed1997

L-selectin ligands in rat high endothelium: multivalent sialyl Lewis x glycans are high-affinity inhibitors of lymphocyte adhesion.

Lymphocyte homing is initiated by their tethering to and rolling on the high endothelium and is followed by extravasation into the lymph nodes. We show here that glycosylated cell adhesion molecule-1 (GlyCAM-1), CD34, and sialyl Lewis x (sLex) are present on rat lymph node high endothelium analyzed by using monoclonal antibodies. alpha (1,3)fucosyltransferase VII (Fuc-TVII), the last enzyme involved in the synthesis of the sLex sequence is also expressed on the rat lymph node high endothelium. We have synthesized a family of sLex-decorated oligosaccharide structures and used them to inhibit lymphocyte binding to high endothelium in the Stamper-Woodruff assay. Monovalent sLex, branched di- and tetravalent sLex, as well as a linear tetravalent sLex significantly reduce lymphocyte binding to endothelium. The branched and linear forms of tetravalent sLex were clearly superior inhibitors of the L-selectin-dependent lymphocyte adhesion, with IC50 values in low nanomolar range. In contrast, the fucose-free analogs having the same charge and approximately the same size as the corresponding sLex glycans had no effect on lymphocyte binding and served as negative controls. Taken together, these data show the crucial importance of sLex in the endothelial ligands for L-selectin. Furthermore, we suggest that L-selectin acts as an oligomer on the lymphocyte surface as it binds multivalent sLex glycans.

Animals↗

PubMed2005

Age-related changes in the hepatic sinusoidal endothelium impede lipoprotein transfer in the rat.

The mechanisms for the association of old age with post-prandial hyperlipidemia and atherosclerosis are not well understood. Post-prandial hyperlipidemia has emerged as a significant risk for atherosclerosis. The liver is the central organ for lipoprotein metabolism. The initial step in the hepatic uptake of post-prandial lipoproteins is their transfer from the hepatic sinusoidal capillary lumen across the hepatic sinusoidal endothelium into the space of Disse. Here, they access hepatocytes for receptor-mediated uptake. We proposed that fenestrations (pores) within the hepatic sinusoidal endothelium filter lipoproteins on the basis of size. Recently we discovered age-related changes in the sinusoidal endothelium (pseudocapillarization), including reduction in the porosity of the endothelium. Using the impulse response technique in perfused rat livers, we found that aging is associated with impaired hepatic transendothelial transfer of chylomicrons with diameters smaller than those of fenestrations. In conclusion, age-related pseudocapillarization of the hepatic sinusoidal endothelium provides a novel mechanism for the association of old age with impaired hepatic lipoprotein metabolism and with atherosclerosis.

Aging↗

PubMed1986

Thrombin uptake and inhibition on endothelium and surfaces with a stable heparin coating: a comparative in vitro study.

The endothelial lining and two differently heparin-coated surfaces were compared in vitro regarding thrombin uptake and inhibition. One heparin surface was based on stabilized ionic binding of heparin, the other on covalent binding of partially degraded heparin. Both heparin surfaces have previously been shown to have pronounced thromboresistant properties. The two heparinized polyethylene surfaces and the endothelial surface of segments of the porcine aorta were studied. After exposure to the surfaces, thrombin disappeared from the solution and appeared bound to the surfaces. The disappearance rate of thrombin from the solution was the same on exposure to the endothelium and the covalently bonded heparin surface, but less following exposure to the ionically bonded heparin surface. The thrombin activity appearing on the endothelium was lower than on the heparin surfaces, indicating that the endothelium exerted a slow thrombin inhibiting capacity. On exposure of the thrombin-loaded endothelium to plasma, thrombin was rapidly inhibited. Thrombin bound to the covalently bonded heparin surface was inhibited at a slower rate than on the ionically bonded surface, but still faster than the rate at which free thrombin was inhibited in nonheparinized plasma. It is concluded that the endothelium and stabilized heparin coatings bind thrombin and accelerate its inhibition by plasma.

Animals↗

PubMed1985

Insulin binding and vesicular ingestion in capillary endothelium.

Capillary endothelium can actively regulate vascular permeability of various serum proteins. Hormones such as insulin must interact with this capillary barrier in order to reach their respective target tissues. We have studied the binding and subsequent internalization of 125I-insulin in both native (freshly isolated) and primary cultured capillary endothelium derived from rat epididymal fat pads. Insulin association with the endothelium, internalization and degradation differed between freshly isolated and primary cultured capillaries. Specific binding in freshly isolated and cultured capillaries was temperature dependent, and was competitively inhibited in the presence of unlabelled insulin. Primary cultures of capillaries grown to confluence did not exhibit specific binding of insulin. Despite the lack of specific receptors for insulin, cultured cells vesicularly internalized insulin. Greater than 50% of the total associated insulin was not degraded by cultured endothelium. Morphological examinations using ferritin labelled insulin localized insulin associated to the capillary endothelial cell membrane and sequestered within pinocytotic vesicles. Incubation of freshly isolated capillaries with insulin stimulated the fluid phase endocytosis of 14C-sucrose; however, insulin had no effect on fluid phase endocytosis in cultured capillaries. These results indicate that capillary endothelium, isolated from rat epididymal fat, exhibit specific receptors for insulin. Binding of insulin to the capillary membrane is followed by internalization into cytoplasmic vesicles and partial degradation.

Animals↗

PubMed1996

Effect of human adrenomedullin on a canine tibial perfusion model in the absence of vascular endothelium.

The effect of human adrenomedullin was investigated using an ex vivo perfused canine tibial model in the absence of vascular endothelium. Adrenomedullin is a novel peptide with known vasodilator properties. In this model, a 0.1 ml bolus injection of 10(-5) M of either acetylcholine or adrenomedullin decreased vascular resistance in tibial preparations precontracted with prostaglandin F2 alpha by 88.3 +/- 3.0% and 92.8 +/- 2.8%, respectively, in the presence of vascular endothelium. Conversely, a 0.1 ml bolus injection of 10(-5) M acetylcholine produced a constrictor response after removal of vascular endothelium. A 0.1 ml bolus injection of 10(-5) M adrenomedullin decreased the baseline perfusion pressure in a dose-dependent manner for a duration of 20 minutes and also attenuated the pressor response to exogenous norepinephrine injection for at least 10 minutes compared with the control study (p < 0.05) in the absence of vascular endothelium. These data suggest that human adrenomedullin relaxes blood vessels in bone by a direct action on vascular smooth-muscle cells. In addition, the attenuation effect of human adrenomedullin on pressor responses to exogenous norepinephrine injection is independent of vascular endothelium.

Acetylcholine↗