Fluid Flow Stimulates Tissue Plasminogen Activator Secretion by Cultured Human Endothelial Cells

Diamond, S. L.; Eskin, S. G.; McIntire, L.V.

doi:10.1126/science.2467379

Cited by 428 publications

(166 citation statements)

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“…These may be later changes, as they are in cultured endothelial cells exposed to shear stress. [3][4][5][6] In only a single condition, nifedipine in the vein perfusate, was significant tissue factor staining observed on endothelium. †Significant increase in tissue factor, (Pϭ0.04).…”

Section: Discussionmentioning

confidence: 99%

“…2 These hemodynamic forces may alter the expression of thrombomodulin, tPA, and tissue factor. [3][4][5][6] In cultured bovine aortic endothelial cells, thrombomodulin protein concentration decreased 3-to 5-fold after 36 hours of exposure to high shear stress, with mRNA levels decreasing as early as 2 to 4 hours after the application of shear stress. 6 In cultured human umbilical endothelial cells, both fluid flow and shear stress have been reported to stimulate mRNA levels and protein secretion of tPA.…”

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confidence: 99%

“…6 In cultured human umbilical endothelial cells, both fluid flow and shear stress have been reported to stimulate mRNA levels and protein secretion of tPA. 4,5 Similarly, although there is scant evidence for the expression of tissue factor in normal endothelium, in cultured cells the application of shear stress causes a transient increase in tissue factor procoagulant activity. 6 In saphenous vein grafted into the arterial circulation, such changes could disturb the antithrombotic properties of endothelium in a manner that may predispose to bypass occlusion.…”

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Arterial Flow Conditions Downregulate Thrombomodulin on Saphenous Vein Endothelium

et al. 1999

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Background —The antithrombogenic properties of venous endothelium may be attenuated when vein is implanted in the arterial circulation. Such changes may facilitate thrombosis, which is the final common pathway for saphenous vein arterial bypass graft occlusion. Methods and Results —Using human saphenous vein in a validated ex vivo flow circuit, we investigated (1) the possibility that arterial flow conditions (mean pressure, 100 mm Hg, 90 cpm, ≈200 mL/min) alter the concentration of proteins involved in regulating thrombosis at the vessel wall and (2) the influence of ion channel blockade on such effects. Concentrations of thrombomodulin and tissue factor were quantified by Western blotting (ratio of von Willebrand factor staining) and immunohistochemistry (as a percentage of CD31-staining area). Thrombomodulin concentrations after 90 minutes of venous and arterial flow conditions were quantified by immunostaining (68.9±4.8% and 41.0±3.0% CD31, respectively; P <0.01) and by Western blotting (1.35±0.20 and 0.15±0.03 ratio of von Willebrand factor, respectively; P <0.01). The ability of endothelial cells to generate activated protein C also decreased from 62±14 to 19±10 ng · min −1 · 1000 cells −1 ( P =0.01). The significant reduction in thrombomodulin was attenuated if calcium was removed from the perfusate but not by external vein stenting. Inclusion in the vein perfusate of drugs that reduce calcium entry (including Gd 3+ , to block stretch-activated ion channels, and nifedipine) abolished the reduction in thrombomodulin concentration observed after arterial flow conditions. In freshly excised vein, negligible concentrations of tissue factor were detected on the endothelium and concentrations did not increase after 90 minutes of arterial flow conditions, although the inclusion of nifedipine caused the immunostaining to increase from 3.0±0.4% to 8.5±0.7% CD31 ( P <0.02). Conclusions —In saphenous vein endothelium exposed to arterial flow conditions, there is rapid downregulation of thrombomodulin, sufficient to limit protein C activation, by a calcium-dependent mechanism.

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Arterial Flow Conditions Downregulate Thrombomodulin on Saphenous Vein Endothelium

et al. 1999

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“…6 Conversely, relative elevations of blood flow and shear stress appear to have an atheroprotective role and inhibit intimal thickening after vascular injury. [7][8][9] Hemodynamic forces have been shown to modulate endothelial and vascular smooth muscle cell (VSMC) responses, which involve the induction of cytokines, such as platelet-derived growth factor (PDGF), basic fibroblast growth factor, and transforming growth factor-␤1 (TGF-␤1), 10,11 in addition to other mediators, such as nitric oxide, 7 tissue plasminogen activator, 12,13 and matrix metalloproteinases (MMPs), 1 4 involved in vascular remodeling.…”

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Increased Flow and Shear Stress Enhance In Vivo Transforming Growth Factor-β1 After Experimental Arterial Injury

Song

Kocharyan

Fortunato

et al. 2000

ATVB

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Abstract-We have previously demonstrated that high-flow (HF) conditions inhibit experimental intimal hyperplasia.We hypothesized that such flow conditions may alter transforming growth factor-␤1 (TGF-␤1) after mural injury. The right common carotid artery (CCA) was balloon-injured in 54 New Zealand White male rabbits. Flow was thereafter preserved (normal flow [NF]), reduced by partial outflow occlusion (low flow [LF]), or increased by ligation of the left CCA (HF). Four sham-operated animals served as uninjured controls. Mean blood flow and pressure in the right CCA were measured before and after flow modulation and before euthanasia (3, 7, and 14 days). TGF-␤1 mRNA and protein levels in the right CCA were determined by Northern and ELISA analyses at each time point. At 7 and 14 days, intimal hyperplasia was quantified, and the transmural localization of TGF-␤1 was determined by immunohistochemical analysis. Mean flow was reduced from 22Ϯ1 to 10Ϯ3 mL/min in the LF group and increased to 34Ϯ2 mL/min in the HF group (PϽ0.001). Blood pressure was not different among the flow groups for all time points. Wall shear stress was markedly decreased in the LF group to 14Ϯ4 dyne/cm 2 and increased in the HF group to 63Ϯ6 dyne/cm 2 at 7 days compared with values in uninjured controls (39Ϯ2 dyne/cm 2 , PϽ0.001) and the NF group (44Ϯ7 dyne/cm 2 , PϽ0.001). At 14 days, wall shear stress was similar among the flow groups. The intima-to-media ratio was 5-and 2-fold greater in the LF group than in the HF and NF groups at 14 days. mRNA levels for TGF-␤1 and its active ligand were increased in the HF group by at least 2-and 3-fold, respectively, at 3 and 7 days compared with levels in uninjured controls and the LF group (PϽ0.05) but were not different among the flow groups at 14 days. TGF-␤1 preferentially localized in the abluminal vascular smooth muscle cells of the HF arterial segments. Flow-and shear-mediated release of TGF-␤1 may therefore play a role in abrogating the proliferative and migratory response of vascular smooth muscle cells in the early stages after mural injury.

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“…Such investigations have demonstrated changes in morphology (Dewey et al 1981), arachidonic acid metabolism (Nollert et al 1989), protein synthesis (Diamond et al 1989), and gene expression (Hsieh et al 1992) in ECs subjected to mechanical shear stress. The available evidence suggests that shear stress stimulates the production of intracellular second messengers rather than causing a general perturbation of cell metabolism.…”

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Effects of Shear Stress on Protein Kinase C Distribution in Endothelial Cells

Chien

1997

J Histochem Cytochem.

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SUMMARYWe studied the effects of shear stress on protein kinase C (PKC) in cultured human umbilical vein endothelial cells by use of a flow channel and a monoclonal antibody (MAb 1.3) that recognizes the PKC ␤ -isozyme. The fluorescence intensity (FI) of the secondary antibody, crystalline tetramethylrhodamine isothiocyanate, was determined by image analysis. The results on each of five shearing experiments were normalized by using the paired stationary control. After 30-min shearing at 2 N/m 2 , FI per cell increased to 1.6 times that of control, as did the mean FI per unit cell area. The FI per unit stained area and the stained area/cell area ratio were also increased significantly by shearing. The distribution of immunostaining in each cell was determined for its cortical, cytoplasmic, perinuclear, and nuclear regions. The normalized FI per unit area in all four regions and the stained area/cell area ratio in cortical and cytoplasmic regions were significantly higher in the sheared cells than in control; the increases were greatest in the cortical area. Double staining with rhodamine-phalloidin and MAb 1.3 showed the association of actin with the PKC isozyme in both stationary and sheared cells.

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Fluid Flow Stimulates Tissue Plasminogen Activator Secretion by Cultured Human Endothelial Cells

Cited by 428 publications

References 34 publications

Arterial Flow Conditions Downregulate Thrombomodulin on Saphenous Vein Endothelium

Arterial Flow Conditions Downregulate Thrombomodulin on Saphenous Vein Endothelium

Increased Flow and Shear Stress Enhance In Vivo Transforming Growth Factor-β1 After Experimental Arterial Injury

Effects of Shear Stress on Protein Kinase C Distribution in Endothelial Cells

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