scholarly journals Evidence against the Hypothesis that Endothelial Endothelin B Receptor Expression Is Regulated by Relaxin and Pregnancy

Endocrinology ◽  
2005 ◽  
Vol 146 (6) ◽  
pp. 2791-2797 ◽  
Author(s):  
Laurie J. Kerchner ◽  
Jacqueline Novak ◽  
Karen Hanley-Yanez ◽  
Ketah D. Doty ◽  
Lee A. Danielson ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Renal Arteries ◽  
Receptor Expression ◽  
Receptor Protein ◽  
Microvascular Endothelial Cells ◽  
Etb Receptor ◽  
Microvascular Endothelial ◽  
Endothelin B

Abstract The endothelial endothelin B (ETB) receptor subtype is critical for renal vasodilation induced by relaxin in nonpregnant rats and during pregnancy (the latter via endogenous circulating relaxin). Here we tested whether expression of vascular ETB receptor protein is regulated by relaxin. Small renal arteries were harvested from virgin and midterm pregnant rats as well as nonpregnant rats that were administered recombinant human relaxin (rhRLX) at 4 μg/h or vehicle for 5 d or 4–6 h. Small renal arteries dissected from additional virgin rats were incubated in vitro with rhRLX or vehicle for 3 h at 37 C. ETB expression was also evaluated in cultured human endothelial cells: aortic, coronary, umbilical vein, and dermal microvascular endothelial cells. Cells were incubated for 4, 8, or 24 h with rhRLX (5, 1, or 0.1 ng/ml) or vehicle. ETB protein expression in arteries and cells was evaluated by Western analysis. No regulation of ETB expression was observed in small renal arteries in any of the experimental protocols, nor was there an increase in the vasorelaxation response to ET-3 in small renal arteries incubated in vitro with rhRLX. rhRLX only sporadically altered ETB expression in human coronary artery endothelial cells and human umbilical vein endothelial cells at certain time points or doses, and no regulation was observed in human aortic endothelial cells or human dermal microvascular endothelial cells. These results suggest that regulation of ETB receptor protein has little or no role in relaxin stimulation of the endothelial ETB/nitric oxide vasodilatory pathway.

scholarly journals CD36 Mediates the In Vitro Inhibitory Effects of Thrombospondin-1 on Endothelial Cells

1997 ◽  
Vol 138 (3) ◽  
pp. 707-717 ◽  
Author(s):  
David W. Dawson ◽  
S. Frieda A. Pearce ◽  
Ruiqin Zhong ◽  
Roy L. Silverstein ◽  
William A. Frazier ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Fusion Proteins ◽  
Solid Phase ◽  
Umbilical Vein ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Microvascular Endothelial Cells ◽  
Thrombospondin 1 ◽  
Microvascular Endothelial

Thrombospondin-1 (TSP-1) is a naturally occurring inhibitor of angiogenesis that is able to make normal endothelial cells unresponsive to a wide variety of inducers. Here we use both native TSP-1 and small antiangiogenic peptides derived from it to show that this inhibition is mediated by CD36, a transmembrane glycoprotein found on microvascular endothelial cells. Both IgG antibodies against CD36 and glutathione-S-transferase–CD36 fusion proteins that contain the TSP-1 binding site blocked the ability of intact TSP-1 and its active peptides to inhibit the migration of cultured microvascular endothelial cells. In addition, antiangiogenic TSP-1 peptides inhibited the binding of native TSP-1 to solid phase CD36 and its fusion proteins, as well as to CD36-expressing cells. Additional molecules known to bind CD36, including the IgM anti-CD36 antibody SM∅, oxidized (but not unoxidized) low density lipoprotein, and human collagen 1, mimicked TSP-1 by inhibiting the migration of human microvascular endothelial cells. Transfection of CD36-deficient human umbilical vein endothelial cells with a CD36 expression plasmid caused them to become sensitive to TSP-1 inhibition of their migration and tube formation. This work demonstrates that endothelial CD36, previously thought to be involved only in adhesion and scavenging activities, may be essential for the inhibition of angiogenesis by thrombospondin-1.

scholarly journals Gui Shao Di Huang Wan promotes angiogenesis and regulates oestrogen receptor α and progesterone receptor β in in vitro bioassays

2021 ◽  
Author(s):  
Rebecca O'Cleirigh ◽  
Rozlyn Gibbs
Keyword(s):  
Endothelial Cells ◽  
Progesterone Receptor ◽  
Mtt Assay ◽  
Receptor Expression ◽  
Microvascular Endothelial Cells ◽  
Ishikawa Cells ◽  
In Vitro Bioassays ◽  
Microvascular Endothelial ◽  
Er Alpha

Background and aim: The formula Gui Shao Di Huang Wan (GSDW) is used frequently to treat female infertility. This study aims to investigate some of the possible mechanisms of action of GSDW using in vitro bioassays of angiogenesis in Human Uterine Microvascular Endothelial Cells (HUVEC) and Human Uterine Microvascular Endothelial Cells (HUtMEC) and ovarian steroid receptor expression in a human endometrial cell line (Ishikawa). Experimental procedure: Aqueous extracts of GSDW and its component herbs were tested for pro-angiogenic activity using both HUVEC and HUtMEC 2D differentiation assays performed on Matrigel and effects on HUVEC proliferation using the MTT assay. Effects on the expression of Estrogen Receptor alpha (ER alpha) and Progesterone Receptor beta (PRbeta) in Ishikawa cells were determined using immunoblotting. Results and Conclusion: ll analysed parameters of differentiation were increased by GSDW in both the HUVEC and HUtMEC mesh. Furthermore, measures of total length, segment number, junction number were affected by some but not all component herbs. The MTT assay showed an increase in proliferation of HUVECs at concentrations of GSDW between 0.68 and 5.47 ug/mL at 48 and 72 hours. In Ishikawa cells downregulation of ER alpha; and upregulation of PR beta; was seen after 48 hours incubation with 4 of the 8 herbs in the formula. The findings in this study demonstrate that GSDW has the potential to affect key parameters (vascular, sex hormone receptor expression) in vitro. This offers a mechanism by which these herbs may enhance fertility through improved endometrial receptivity and pregnancy rates.

scholarly journals On-chip perivascular niche with patient-derived glioma cells

2020 ◽  
Author(s):  
Magda Gerigk ◽  
Harry Bulstrode ◽  
HaoTian Harvey Shi ◽  
Felix Tönisen ◽  
Camilla Cerutti ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
3D Culture ◽  
Glioma Cells ◽  
Microvascular Endothelial Cells ◽  
Normal Brain ◽  
Tumour Mass ◽  
Perivascular Niche ◽  
Microvascular Endothelial

AbstractGlioblastoma multiforme (GBM), is the most common and the most aggressive type of primary brain malignancy. Glioblastoma stem-like cells (GSCs) are able to migrate in vascular niches within or away from the tumour mass, increasing tumour resistance to patient treatments and contributing to relapses. To study individual GSCs migration and their interactions with the microenvironment in the vasculature, there is a need to develop a model of human blood vessels in vitro. Herein, we report a systematic study on the interaction between patient-derived glioma stem-like cell lines with different organotypic perivascular niche models. A microfluidic chip integrated with an extracellular matrix was fabricated to support the culture of rounded microvessels, formed with endothelial cells from three different organs, (1) human brain microvascular endothelial cells (hCMEC/D3), (2) human umbilical vein endothelial cells (HUVECs) and, (3) human lung microvascular endothelial cells (HMVEC-L). Three-dimensional (3D) cell culture retains selected adherent and tight junction markers of the endothelial cells, and the stemness-related genes of GSCs. We optimized the experimental protocol to perform qPCR, and western blot on the co-cultured GSCs with endothelial cells forming microvessels. Endpoint biological assays showed upregulation of neovascularization-related genes in endothelial cells (e.g., angiopoietins, vascular endothelial growth factor receptors) resulted after their co-culture with GBM cells. Moreover, we measured cancer cell speed and polarization during migration towards the endothelial cell formed vessel by live-cell imaging showing that organotypic (brain cancer cells – brain endothelial microvessel) interactions differ from those within non-tissue specific vascular niches. The development and optimization of this 3D microfluidic device could provide the next level of complexity of an in vitro system to study the influence of glioma cells on normal brain endothelium. More importantly, it enables the possibility to conduct comparative studies to dissect the influence of 3D culture, microvessel architecture and organotypic vessel types on glioma cells’ stemness and migration.

C-C Motif Chemokine 8 promotes angiogenesis in vascular endothelial cells

Vascular ◽  
2020 ◽  
pp. 170853812095997
Author(s):  
Song Xue ◽  
Hanfei Tang ◽  
Gefei Zhao ◽  
Yang Shen ◽  
Ethan Yibo Yang ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Map Kinase ◽  
Chemokine Receptor ◽  
Umbilical Vein ◽  
Tube Formation ◽  
Microvascular Endothelial Cells ◽  
Human Umbilical Vein ◽  
Microvascular Endothelial ◽  
Endothelium Cells

Objectives Angiogenesis is an important progress associated with several pathological situations. Several chemokines have been reported to act as regulators of angiogenesis. The current study aimed to find whether C-C Motif Chemokine 8 is involved in angiogenesis regulation. Methods To verify whether C-C Motif Chemokine 8 is related to angiogenesis in plaques, carotid plaques were collected from patients with severe carotid stenosis and analysed using CD31 immunohistochemistry and real-time PCR. To further clarify the relation between C-C Motif Chemokine 8 and angiogenesis, human umbilical vein endothelium cells and human dermal microvascular endothelial cells were treated with C-C Motif Chemokine 8 in the presence or absence of C-C motif chemokine receptor 2-Ab and extracellular regulated MAP kinase 1/2 inhibition (FR180204). Proliferation and migration of human umbilical vein endothelium cells and human dermal microvascular endothelial cells were examined with Cell Counting Kit-8 and Transwell chamber assay, respectively. In vitro angiogenesis stimulated by C-C Motif Chemokine 8 was examined using tube formation assay. Ex vivo and in vivo angiogenesis were assessed by mice aortic ring assay and Matrigel plug assay, respectively. C-C motif chemokine receptors of human umbilical vein endothelium cells were examined with real-time PCR, and C-C motif chemokine receptor 1, C-C motif chemokine receptor 2, extracellular regulated MAP kinase 1/2 and phosphorylation-extracellular regulated MAP kinase 1/2 were examined with western blotting assay. Results C-C Motif Chemokine 8 was increased in carotid plaques with severe angiogenesis in both RNA and protein level. C-C Motif Chemokine 8 (5 ng/ml) weakly increased human umbilical vein endothelium cell proliferation, but not on human dermal microvascular endothelial cells. Migration and tube formation could be induced by C-C Motif Chemokine 8 in both human umbilical vein endothelium cells and human dermal microvascular endothelial cells. In mice aortic ring assay and Matrigel plug assay, C-C Motif Chemokine 8 could promote angiogenesis compared to vehicle groups. Phosphorylation of extracellular regulated MAP kinase 1/2 was increased with C-C Motif Chemokine 8 stimulation. The migration and tube formation promoted by C-C Motif Chemokine 8 could be largely blocked by C-C motif chemokine receptor 2-Ab or extracellular regulated MAP kinase 1/2 inhibition (FR180204). Conclusions C-C Motif Chemokine 8 could promote both in vitro and in vivo angiogenesis. C-C motif chemokine receptor 2 played an important role in the activation of C-C Motif Chemokine 8 and extracellular regulated MAP kinase 1/2 signalling pathway was involved in this mechanism.

Isolation and primary culture of rat cerebral microvascular endothelial cells for studying drug transport in vitro

1996 ◽  
Vol 36 (1) ◽  
pp. 45-52 ◽  
Author(s):  
Nobuhiro Ichikawa ◽  
Kohji Naora ◽  
Hidenari Hirano ◽  
Michio Hashimoto ◽  
Sumio Masumura ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Primary Culture ◽  
Drug Transport ◽  
Microvascular Endothelial Cells ◽  
Microvascular Endothelial

scholarly journals Human Brain Microvascular Endothelial Cells and Umbilical Vein Endothelial Cells Differentially Facilitate Leukocyte Recruitment and Utilize Chemokines for T Cell Migration

2008 ◽  
Vol 2008 ◽  
pp. 1-8 ◽  
Author(s):  
Shumei Man ◽  
Eroboghene E. Ubogu ◽  
Katherine A. Williams ◽  
Barbara Tucky ◽  
Melissa K. Callahan ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Migration ◽  
T Cell ◽  
Human Brain ◽  
Umbilical Vein ◽  
Microvascular Endothelial Cells ◽  
Brain Microvascular Endothelial Cells ◽  
T Cell Migration ◽  
Umbilical Vein Endothelial Cells

Endothelial cells that functionally express blood brain barrier (BBB) properties are useful surrogates for studying leukocyte-endothelial cell interactions at the BBB. In this study, we compared two different endothelial cellular models: transfected human brain microvascular endothelial cells (THBMECs) and human umbilical vein endothelial cells (HUVECs). With each grow under optimal conditions, confluent THBMEC cultures showed continuous occludin and ZO-1 immunoreactivity, while HUVEC cultures exhibited punctate ZO-1 expression at sites of cell-cell contact only. Confluent THBMEC cultures on 24-well collagen-coated transwell inserts had significantly higher transendothelial electrical resistance (TEER) and lower solute permeability than HUVECs. Confluent THBMECs were more restrictive for mononuclear cell migration than HUVECs. Only THBMECs utilized abluminal CCL5 to facilitate T-lymphocyte migration in vitro although both THBMECs and HUVECs employed CCL3 to facilitate T cell migration. These data establish baseline conditions for using THBMECs to develop in vitro BBB models for studying leukocyte-endothelial interactions during neuroinflammation.

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