scholarly journals Perfusion fixation method is critical for immunoelectron microscopy and ultrastructural evaluation on changes of caveolin-1 and caveolae relates with capillarization of liver sinusoidal endothelial cells in human cirrhotic liver

2014 ◽  
Vol 7 (1) ◽  
pp. 32-32
Author(s):  
Hiroaki Yokomori ◽  
Jing-Yan Han ◽  
Masaya Oda
Keyword(s):  
Endothelial Cells ◽  
Immunoelectron Microscopy ◽  
Cirrhotic Liver ◽  
Fixation Method ◽  
Liver Sinusoidal Endothelial Cells ◽  
Sinusoidal Endothelial Cells ◽  
Caveolin 1 ◽  
Perfusion Fixation

scholarly journals oxLDL induces injury and defenestration of human liver sinusoidal endothelial cells via LOX1

2014 ◽  
Vol 53 (2) ◽  
pp. 281-293 ◽  
Author(s):  
Qi Zhang ◽  
Jing Liu ◽  
Jia Liu ◽  
Wenhui Huang ◽  
Limin Tian ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Human Liver ◽  
Underlying Mechanism ◽  
Ros Generation ◽  
Dependent Manner ◽  
Liver Sinusoidal Endothelial Cells ◽  
Sinusoidal Endothelial Cells ◽  
Alcoholic Fatty Liver ◽  
Caveolin 1

Non-alcoholic fatty liver disease is associated with hepatic microangiopathy and liver inflammation caused by type 2 diabetes mellitus. Oxidised LDL (oxLDL) is involved in proinflammatory and cytotoxic events in various microcirculatory systems. The lectin-like oxLDL receptor 1 (LOX1) plays a crucial role in oxLDL-induced pathological transformation. However, the underlying mechanism of oxLDL's effects on liver microcirculation disturbances remains unclear. In this study, we investigated the effects of oxLDL on LOX1 (OLR1) expression and function, as well as on the fenestration features of human liver sinusoidal endothelial cells (HLSECs)in vitro. Primary HLSECs were obtained and cultured. The cells were treated with various concentrations of oxLDL (25, 50, 100 and 200 μg/ml), and the cytotoxicity and expression of LOX1 were examined. Furthermore,LOX1knockdown was performed using siRNA technology, and the changes in intracellular reactive oxygen species (ROS), NFκB, p65, (p65), endothelin 1 (ET1 (EDN1)), eNOS (NOS3) and caveolin 1 (CAV1) levels were measured. Cells were treated with 100 μg/ml oxLDL, and the fenestra morphology was visualised using scanning electron microscopy. oxLDL significantly increased LOX1 expression at both the mRNA and protein levels in HLSECs in a dose- and time-dependent manner. oxLDL stimulation increased ROS generation and NFκB activation, upregulated ET1 and caveolin 1 expression, downregulated eNOS expression and reduced the fenestra diameter and porosity. All of these oxLDL-mediated effects were inhibited afterLOX1knockdown. These results reveal a mechanism by which oxLDL stimulates the production of LOX1 through the ROS/NFκB signalling pathway and by which LOX1 mediates oxLDL-induced endothelial injury and the defenestration of HLSECs.

scholarly journals The response of fenestrations, actin, and caveolin-1 to vascular endothelial growth factor in SK Hep1 cells

2008 ◽  
Vol 295 (1) ◽  
pp. G137-G145 ◽  
Author(s):  
Victoria C. Cogger ◽  
Irwin M. Arias ◽  
Alessandra Warren ◽  
Aisling C. McMahon ◽  
Debra L. Kiss ◽  
...  
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Endothelial Cells ◽  
Growth Factor ◽  
Growth Factor Receptor ◽  
Green Fluorescence Protein ◽  
Vascular Endothelial ◽  
Liver Sinusoidal Endothelial Cells ◽  
Sinusoidal Endothelial Cells ◽  
Caveolin 1 ◽  
Endothelial Growth Factor

To study the regulation of fenestrations by vascular endothelial growth factor in liver sinusoidal endothelial cells, SK Hep1 cells were transfected with green fluorescence protein (GFP)-actin and GFP-caveolin-1. SK Hep1 cells had pores; some of which appeared to be fenestrations (diameter 55 ± 28 nm, porosity 2.0 ± 1.4%), rudimentary sieve plates, bristle-coated micropinocytotic vesicles and expressed caveolin-1, von Willebrand factor, vascular endothelial growth factor receptor-2, endothelial nitric oxide synthase and clathrin, but not CD31. There was avid uptake of formaldehyde serum albumin, consistent with endocytosis. Vascular endothelial growth factor caused an increase in porosity to 4.8 ± 2.6% ( P < 0.01) and pore diameter to 104 ± 59 nm ( P < 0.001). GFP-actin was expressed throughout the cells, whereas GFP-caveolin-1 had a punctate appearance; both responded to vascular endothelial growth factor by contraction toward the nucleus over hours in parallel with the formation of fenestrations. SK Hep1 cells resemble liver sinusoidal endothelial cells, and the vascular endothelial growth factor-induced formation of fenestration-like pores is preceded by contraction of actin cytoskeleton and attached caveolin-1 toward the nucleus.

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