scholarly journals Simple, Reliable Isolation, Purification and Cultivation of Murine Skeletal Muscle Microvascular Endothelial Cells

2020 ◽  
Vol 13 (12) ◽  
pp. 290-299
Author(s):  
Jianjie Wang ◽  
Joseph Harvey ◽  
Richard Garrad ◽  
Virginia Huxley ◽  
Laurie Erb ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Endothelial Cells ◽  
Microvascular Endothelial Cells ◽  
Microvascular Endothelial ◽  
Murine Skeletal Muscle ◽  
Reliable Isolation

Isolation of Primary Murine Skeletal Muscle Microvascular Endothelial Cells

10.3791/58901 ◽  
2019 ◽  
Author(s):  
Thomas Müntefering ◽  
Alexander P.E. Michels ◽  
Steffen Pfeuffer ◽  
Sven G. Meuth ◽  
Tobias Ruck
Keyword(s):  
Skeletal Muscle ◽  
Endothelial Cells ◽  
Microvascular Endothelial Cells ◽  
Microvascular Endothelial ◽  
Murine Skeletal Muscle

Vasoconstrictor effects of insulin in skeletal muscle arterioles are mediated by ERK1/2 activation in endothelium

2004 ◽  
Vol 287 (5) ◽  
pp. H2043-H2048 ◽  
Author(s):  
Etto C. Eringa ◽  
Coen D. A. Stehouwer ◽  
Geerten P. van Nieuw Amerongen ◽  
Lenneke Ouwehand ◽  
Nico Westerhof ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Endothelial Cells ◽  
Fold Increase ◽  
Pi3 Kinase ◽  
Glucose Disposal ◽  
Microvascular Endothelial Cells ◽  
Kinase Inhibition ◽  
Basal Diameter ◽  
Extracellular Signal ◽  
Microvascular Endothelial

Insulin exerts both NO-dependent vasodilator and endothelin-dependent vasoconstrictor effects on skeletal muscle arterioles. The intracellular enzymes 1-phosphatidylinositol 3-kinase (PI3-kinase) and Akt have been shown to mediate the vasodilator effects of insulin, but the signaling molecules involved in the vasoconstrictor effects of insulin in these arterioles are unknown. Our objective was to identify intracellular mediators of acute vasoconstrictor effects of insulin on skeletal muscle arterioles. Rat cremaster first-order arterioles ( n = 40) were isolated, and vasoreactivity to insulin was studied using a pressure myograph. Insulin induced dose-dependent vasoconstriction of skeletal muscle arterioles (up to −22 ± 3% of basal diameter; P < 0.05) during PI3-kinase inhibition with wortmannin (50 nmol/l). Insulin-induced vasoconstriction was abolished by inhibition of extracellular signal-regulated kinase 1/2 (ERK1/2) with PD-98059 (40 μmol/l). In addition, inhibition of ERK1/2 without PI3-kinase inhibition uncovered insulin-mediated vasodilatation in skeletal muscle arterioles (up to 37 ± 10% of baseline diameter; P < 0.05). Effects of insulin on ERK1/2 activation in arterioles were then investigated by Western blot analysis. Insulin induced a transient 2.4-fold increase in ERK1/2 phosphorylation (maximal at ∼15 min) in skeletal muscle arterioles ( P < 0.05). Removal of the arteriolar endothelium abolished insulin-induced vasoconstriction, which suggests that activation of ERK1/2 in endothelial cells is involved in acute insulin-mediated vasoconstriction. To investigate this, acute effects of insulin on ERK1/2 phosphorylation were studied in human microvascular endothelial cells. In support of the findings in skeletal muscle arterioles, insulin induced a 1.9-fold increase in ERK1/2 phosphorylation (maximal at ∼15 min) in microvascular endothelial cells ( P < 0.05). We conclude that acute vasoconstrictor effects of insulin in skeletal muscle arterioles are mediated by activation of ERK1/2 in endothelium. This ERK1/2-mediated vasoconstrictor effect antagonizes insulin-induced, PI3-kinase-dependent vasodilatation in skeletal muscle arterioles. These findings provide a novel mechanism by which insulin may determine blood flow and glucose disposal in skeletal muscle.

Ascorbate Uptake by Microvascular Endothelial Cells of Rat Skeletal Muscle

1996 ◽  
Vol 3 (2) ◽  
pp. 211-221 ◽  
Author(s):  
J. X. Wilson ◽  
S. J. Dixon ◽  
J. Yu ◽  
S. Nees ◽  
K. Tyml
Keyword(s):  
Skeletal Muscle ◽  
Endothelial Cells ◽  
Microvascular Endothelial Cells ◽  
Rat Skeletal Muscle ◽  
Microvascular Endothelial ◽  
Ascorbate Uptake

Resveratrol Reduces Matrix Metalloproteinase-2 Activity Induced by Oxygen-Glucose Deprivation and Reoxygenation in Human Cerebral Microvascular Endothelial Cells

2012 ◽  
Vol 82 (4) ◽  
pp. 267-274 ◽  
Author(s):  
Zahide Cavdar ◽  
Mehtap Y. Egrilmez ◽  
Zekiye S. Altun ◽  
Nur Arslan ◽  
Nilgun Yener ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Neuroprotective Effect ◽  
Ischemia Reperfusion ◽  
Neurovascular Unit ◽  
Glucose Deprivation ◽  
Matrix Metalloproteinase 2 ◽  
Microvascular Endothelial Cells ◽  
Oxygen Glucose Deprivation ◽  
Microvascular Endothelial

The main pathophysiology in cerebral ischemia is the structural alteration in the neurovascular unit, coinciding with neurovascular matrix degradation. Among the human matrix metalloproteinases (MMPs), MMP-2 and -9, known as gelatinases, are the key enzymes for degrading type IV collagen, which is the major component of the basal membrane that surrounds the cerebral blood vessel. In the present study, we investigated the effects of resveratrol on cytotoxicity, reactive oxygen species (ROS), and gelatinases (MMP-2 and -9) in human cerebral microvascular endothelial cells exposed to 6 hours of oxygen-glucose deprivation and a subsequent 24 hours of reoxygenation with glucose (OGD/R), to mimic ischemia/reperfusion in vivo. Lactate dehydrogenase increased significantly, in comparison to that in the normoxia group. ROS was markedly increased in the OGD/R group, compared to normoxia. Correspondingly, ROS was significantly reduced with 50 μM of resveratrol. The proMMP-2 activity in the OGD/R group showed a statistically significant increase from the control cells. Resveratrol preconditioning decreased significantly the proMMP-2 in the cells exposed to OGD/R in comparison to that in the OGD/R group. Our results indicate that resveratrol regulates MMP-2 activity induced by OGD/R via its antioxidant effect, implying a possible mechanism related to the neuroprotective effect of resveratrol.

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