scholarly journals Severe sepsis cytomix elicits inflammation in cerebrovascular endothelial cells and polymorphonuclear (PMN) leukocytes in vitro.

2013 ◽  
Vol 27 (S1) ◽  
Author(s):  
Christopher Blom ◽  
Brittany Deller ◽  
Douglas D Fraser ◽  
Claudio Martin ◽  
Bryan Young ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Severe Sepsis ◽  
Pmn Leukocytes ◽  
Cerebrovascular Endothelial Cells

Abstract 195: Simvastatin Rescues Impaired Angiogenesis Potential of Cerebrovascular Endothelial Cells by AGEs : The Role of Autophagy

Stroke ◽  
2014 ◽  
Vol 45 (suppl_1) ◽  
Author(s):  
Xuwei Hou ◽  
Zhaohui Hu ◽  
Ningfu Wang
Keyword(s):  
Endothelial Cells ◽  
Tube Formation ◽  
Western Blot Assay ◽  
Angiogenic Potential ◽  
Cellular Autophagy ◽  
Glycation End Products ◽  
Angiogenic Effect ◽  
Cerebrovascular Endothelial Cells ◽  
High Level

Background: Statin therapy is beneficial for ischemic stroke (IS) partly due to its pro-angiogenic effect. In diabetes patients, the accumulations of advanced glycation end products (AGEs) impair the angiogenic potential of endothelial cells (ECs), comprising collateral circulation in ischemic area. Autophagy affects endothelial function. The effect of statins on EC autophagy remains largely unknown. This study was to: 1.investigate the effect of Statin on the angiogenesis ability of ECs impaired by AGEs; 2. explore the autophagy of ECs in this process. Methods: primary cerebromicrovascular endothelial cells (CMVECs) were isolated from Wistar rats and cultured in medium. CMVECs were treated with AGEs. Subsequently, Simvastatin was added to cells. The angiogenic potential was evaluated by tube formation in vitro. Two autophagy marker, Beclin1 and LC3 protein expression were detected by western blot assay. The autophagosome quantitative analysis was evaluated by GFP-LC3-II assay. Results: AGEs (25mg/L, 50mg/L and 100mg/L) reduced the number of tube like structures in a dose dependant manner, accompanied by dramatically enhanced cellular autophagy level indicated by the increased GFP-LC3-II puncta number/cell and increased Beclin1 and LC3 expressions. Autophagy inhibitor 3-MA reversed the AGEs impaired angiogenic ability of CMVES. Simvastatin administration (10μmol/L) markedly increased the angiogenic potential of CMVES evaluated by tube formation ability, together with decreased Beclin1 and LC3, as well as decreased GFP-LC3-II puncta number/cell. AGEs significantly inhibits the PI3K/Akt/mTOR, while Simvastatin re-activate this pathway. More interestingly, the addition of mTOR inhibitor, rapamycin, which enhance the cellular autophagy, abolished the rescue effect of Simvastatin on the angiogenic potential of CMVECs impaired by AGEs. Conclusion: This study provides evidence that AGEs impair the angiogenesis ability via triggering high level of autophagy of CMVECs. Simvastatin rescue the angiogenic ability of CMVECs by surprising autophagy level via activation of PI3K/Akt/mTOR pathway.

scholarly journals Structural, Functional, and Metabolic Alterations in Human Cerebrovascular Endothelial Cells during Toxoplasma gondii Infection and Amelioration by Verapamil In Vitro

2020 ◽  
Vol 8 (9) ◽  
pp. 1386
Author(s):  
Alaa T. Al-sandaqchi ◽  
Victoria Marsh ◽  
Huw E. L. Williams ◽  
Carl W. Stevenson ◽  
Hany M. Elsheikha
Keyword(s):  
Endothelial Cells ◽  
Toxoplasma Gondii ◽  
Intercellular Junctions ◽  
Proton Nuclear Magnetic Resonance ◽  
Functional Changes ◽  
Brain Infection ◽  
Toxoplasma Gondii Infection ◽  
Cerebrovascular Endothelial Cells ◽  
Treatment Application

Toxoplasma gondii (T. gondii), the causative agent of toxoplasmosis, is a frequent cause of brain infection. Despite its known ability to invade the brain, there is still a dire need to better understand the mechanisms by which this parasite interacts with and crosses the blood–brain barrier (BBB). The present study revealed structural and functional changes associated with infection and replication of T. gondii within human brain microvascular endothelial cells (BMECs) in vitro. T. gondii proliferated within the BMECs and disrupted the integrity of the cerebrovascular barrier through diminishing the cellular viability, disruption of the intercellular junctions and increasing permeability of the BMEC monolayer, as well as altering lipid homeostasis. Proton nuclear magnetic resonance (1H NMR)-based metabolomics combined with multivariate data analysis revealed profiles that can be attributed to infection and variations in the amounts of certain metabolites (e.g., amino acids, fatty acids) in the extracts of infected compared to control cells. Notably, treatment with the Ca2+ channel blocker verapamil rescued BMEC barrier integrity and restricted intracellular replication of the tachyzoites regardless of the time of treatment application (i.e., prior to infection, early- and late-infection). This study provides new insights into the structural and functional changes that accompany T. gondii infection of the BMECs, and sheds light upon the ability of verapamil to inhibit the parasite proliferation and to ameliorate the adverse effects caused by T. gondii infection.

scholarly journals Cytosolic Ca2+ Oscillations in Human Cerebrovascular Endothelial Cells after Subarachnoid Hemorrhage

2008 ◽  
Vol 29 (1) ◽  
pp. 57-65 ◽  
Author(s):  
Wolfram Scharbrodt ◽  
Yaser Abdallah ◽  
Sascha A Kasseckert ◽  
Dragan Gligorievski ◽  
Hans M Piper ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Subarachnoid Hemorrhage ◽  
Cerebral Vasospasm ◽  
In Vitro Model ◽  
Molecular Mechanisms ◽  
Vascular Inflammation ◽  
Cell Contraction ◽  
Vitro Model ◽  
Cerebrovascular Endothelial Cells

Molecular mechanisms of cerebral vasospasm after subarachnoid hemorrhage (SAH) include specific modes of cell signaling like activation of nuclear factor (NF)-kB and vascular cell adhesion molecules (VCAM)-1 expression. The study's hypothesis is that cisternal cerebral spinal fluid (CSF) from patients after SAH may cause Ca2+ oscillations which induce these modes of vascular inflammation in an in vitro model of human cerebral endothelial cells (HCECs). HCECs were incubated with cisternal CSF from 10 SAH patients with confirmed cerebral vasospasm. The CSF was collected on days 5 and 6 after hemorrhage. Cytosolic Ca2+ concentrations and cell contraction as an indicator of endothelial barrier function were examined by fura-2 microflurometry. Activation of NF-κB and VCAM-1 expression were measured by immunocytochemistry. Incubation of HCEC with SAH-CSF provoked cytosolic Ca2+ oscillations (0.31 ± 0.09 per min), cell contraction, NF-κB activation, and VCAM-1 expression, whereas exposure to native CSF had no significant effect. When endoplasmic reticulum (ER) Ca2+-ATPase and ER inositol trisphosphate (IP3)-sensitive Ca2+ channels were blocked by thapsigargin or xestospongin, the frequency of the Ca2+ oscillations was reduced significantly. In analogy to the reduction of Ca2+ oscillation frequency, the blockers impaired HCEC contraction, NF-κB activation, and VCAM-1 expression. Cisternal SAH-CSF induces cytosolic Ca2+ oscillations in HCEC that results in cellular constriction, NF-κB activation, and VCAM-1 expression. The Ca2+ oscillations depend on the function of ER Ca2+-ATPase and IP3-sensitive Ca2+ channels.

Ultrastructural studies on the interaction of haemophilus influenzae with human endothelial cells in vitro

1990 ◽  
Vol 48 (3) ◽  
pp. 636-637
Author(s):  
D.J.P. Ferguson ◽  
M. Virji ◽  
H. Kayhty ◽  
E.R. Moxon
Keyword(s):  
Endothelial Cells ◽  
Haemophilus Influenzae ◽  
Intercellular Junctions ◽  
Blood Stream ◽  
Ultrastructural Studies ◽  
Endothelial Barriers ◽  
Serotype B ◽  
Meningitis In Children ◽  
Respiratory Epithelial

Haemophilus influenzae is a human pathogen which causes meningitis in children. Systemic H. influenzae infection is largely confined to encapsulated serotype b organisms and is a major cause of meningitis in the U.K. and elsewhere. However, the pathogenesis of the disease is still poorly understood. Studies in the infant rat model, in which intranasal challenge results in bacteraemia, have shown that H. influenzae enters submucosal tissues and disseminates to the blood stream within minutes. The rapidity of these events suggests that H. influenzae penetrates both respiratory epithelial and endothelial barriers with great efficiency. It is not known whether the bacteria penetrate via the intercellular junctions, are translocated within the cells or carried across the cellular barrier in 'trojan horse' fashion within phagocytes. In the present studies, we have challenged cultured human umbilical cord_vein endothelial cells (HUVECs) with both capsulated (b+) and capsule-deficient (b-) isogenic variants of one strain of H. influenzae in order to investigate the interaction between the bacteria and HUVEC and the effect of the capsule.

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