scholarly journals Eucalyptol Alleviates Amyloid-β-Responsive ER Stress-Induced Endothelial Apoptosis in Diabetic Eyes

2021 ◽  
Vol 5 (Supplement_2) ◽  
pp. 333-333
Author(s):  
Dongyeon Kim ◽  
Min-Kyung Kang ◽  
Young-Hee Kang
Keyword(s):  
Endothelial Cells ◽  
Er Stress ◽  
Apoptotic Cell ◽  
Amyloid Β ◽  
Microvascular Endothelial Cells ◽  
Funding Sources ◽  
Diabetic Retina ◽  
Microvascular Endothelial ◽  
The Unfolded Protein Response

Abstract Objectives Prolonged endoplasmic reticulum (ER) stress impairs the protein folding capacity, and the accumulation of unfolded proteins in the ER eventually triggers apoptosis. This study was aimed to explore that amyloid-β (Aβ) deposited in the diabetic retina, induced retinal endothelial ER stress and apoptosis, which was attenuated by eucalyptol, a monoterpenoid and an achiral aromatic component of many plants, including Salvia and Eucalyptus leaves. Methods Primary human retinal microvascular endothelial cells (HRMVEC) were cultured in media containing 33 mM glucose and 5 μM amyloid-β for 3 days in the presence of 1–20 μM eucalyptol. The in vivo animal study employed db/db mice orally administrated with 10 mg/kg eucalyptol. Cell lysates and mouse eye tissue extracts were prepared for Western blotting, in which antibodies of Aβ, phospho-PERK, phospho-elF2α, ATF4, CHOP and Bax were used. Results Hyperglycemia enhanced the induction of Aβ protein in microvascular endothelial cells, which was inhibited by eucalyptol. Exposure of HRMVEC to high glucose and Aβ elevated the unfolded protein response to ER stress, evidenced by activation of PERK-elF2α-ATF4-CHOP. Eucalyptol diminished formation of glucose- and Aβ-induced ER stress and induction of apoptosis-related bax. Consistently, oral administration of eucalyptol to db/db mice curtailed the eye tissue levels of ER-responsive components and apoptotic proteins. Conclusions Eucalyptol may be a potent retinoprotective agent allaying diabetes-mediated Aβ-induced ER stress leading to apoptotic cell death. Funding Sources This work was supported by the National Research Foundation of Korea (NRF) grants funded by the Korea government (2020R1A6A3A13069116 and 2017R1A6A3A04011473) and was supported by the BK21 FOUR(Fostering Outstanding Universities for Research, 4220200913807) funded by the National Research Foundation of Korea (NRF).

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.

scholarly journals 2-Chlorohexadecanoic acid induces ER stress and mitochondrial dysfunction in brain microvascular endothelial cells

Redox Biology ◽  
2018 ◽  
Vol 15 ◽  
pp. 441-451 ◽  
Author(s):  
Eva Bernhart ◽  
Nora Kogelnik ◽  
Jürgen Prasch ◽  
Benjamin Gottschalk ◽  
Madeleine Goeritzer ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Er Stress ◽  
Mitochondrial Dysfunction ◽  
Microvascular Endothelial Cells ◽  
Brain Microvascular Endothelial Cells ◽  
Microvascular Endothelial

scholarly journals Application of Signature-Tagged Mutagenesis for Identification ofEscherichia coli K1 Genes That Contribute to Invasion of Human Brain Microvascular Endothelial Cells

2000 ◽  
Vol 68 (9) ◽  
pp. 5056-5061 ◽  
Author(s):  
Julie L. Badger ◽  
Carol A. Wass ◽  
Scott J. Weissman ◽  
Kwang Sik Kim
Keyword(s):  
Endothelial Cells ◽  
Human Brain ◽  
Neonatal Rat ◽  
Microvascular Endothelial Cells ◽  
Selection Strategy ◽  
Bacterial Invasion ◽  
Brain Microvascular Endothelial Cells ◽  
E Coli ◽  
Microvascular Endothelial

ABSTRACT Escherichia coli K1 is the leading cause of gram-negative bacterial meningitis in neonates. It is principally due to our limited understanding of the pathogenesis of this disease that the morbidity and mortality rates remain unacceptably high. To identify genes required for E. coli K1 penetration of the blood-brain barrier (BBB), we used the negative selection strategy of signature-tagged transposon mutagenesis (STM) to screen mutants for loss or decreased invasion of human brain microvascular endothelial cells (HBMEC) which comprise the BBB. A total of 3,360 insertion mutants of E. coli K1 were screened, and potential HBMEC invasion mutants were subjected to a secondary invasion screen. Those mutants that failed to pass the serial invasion screens were then tested individually. Seven prototrophic mutants were found to exhibit significantly decreased invasive ability in HBMEC. We identifiedtraJ and five previously uncharacterized loci whose gene products are necessary for HBMEC invasion by E. coli K1. In addition, cnf1, a gene previously shown to play a role in bacterial invasion, was identified. More importantly, atraJ mutant was attenuated in penetration of the BBB in the neonatal rat model of experimental hematogenous meningitis. This is the first in vivo demonstration that traJ is involved in the pathogenesis of E. coli K1 meningitis.

scholarly journals Tranilast inhibits the proliferation, chemotaxis and tube formation of human microvascular endothelial cells in vitro and angiogenesis in vivo

1997 ◽  
Vol 122 (6) ◽  
pp. 1061-1066 ◽  
Author(s):  
Masayuki Isaji ◽  
Hiroshi Miyata ◽  
Yoshiyuki Ajisawa ◽  
Yasuo Takehana ◽  
Nagahisa Yoshimura
Keyword(s):  
Endothelial Cells ◽  
Tube Formation ◽  
Microvascular Endothelial Cells ◽  
Microvascular Endothelial

scholarly journals A Genomic Comparison of in vivo and in vitro Brain Microvascular Endothelial Cells

2007 ◽  
Vol 28 (1) ◽  
pp. 135-148 ◽  
Author(s):  
Anthony R Calabria ◽  
Eric V Shusta
Keyword(s):  
Endothelial Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Genomic Analysis ◽  
Subtractive Hybridization ◽  
Gene Panel ◽  
Microvascular Endothelial Cells ◽  
Brain Microvascular Endothelial Cells ◽  
Microvascular Endothelial

The blood—brain barrier (BBB) is composed of uniquely differentiated brain microvascular endothelial cells (BMEC). Often, it is of interest to replicate these attributes in the form of an in vitro model, and such models are widely used in the research community. However, the BMEC used to create in vitro BBB models de-differentiate in culture and lose many specialized characteristics. These changes are poorly understood at a molecular level, and little is known regarding the consequences of removing BMEC from their local in vivo microenvironment. To address these issues, suppression subtractive hybridization (SSH) was used to identify 25 gene transcripts that were differentially expressed between in vivo and in vitro BMEC. Genes affected included those involved in angiogenesis, transport and neurogenesis, and real-time quantitative polymerase chain reaction (qPCR) verified transcripts were primarily and significantly downregulated. Since this quantitative gene panel represented those BMEC characteristics lost upon culture, we used it to assess how culture manipulation, specifically BMEC purification and barrier induction by hydrocortisone, influenced the quality of in vitro models. Puromycin purification of BMEC elicited minimal differences compared with untreated BMEC, as assessed by qPCR. In contrast, qPCR-based gene panel analysis after induction with hydrocortisone indicated a modest shift of 10 of the 23 genes toward a more ‘ in vivo-like’ gene expression profile, which correlated with improved barrier phenotype. Genomic analysis of BMEC de-differentiation in culture has thus yielded a functionally diverse set of genes useful for comparing the in vitro and in vivo BBB.

scholarly journals Basal lamina formation by cultured microvascular endothelial cells.

1984 ◽  
Vol 99 (2) ◽  
pp. 692-698 ◽  
Author(s):  
R H Kramer ◽  
K G Bensch ◽  
P M Davison ◽  
M A Karasek
Keyword(s):  
Endothelial Cells ◽  
Basal Lamina ◽  
Experimental System ◽  
Microvascular Endothelial Cells ◽  
Type Iv ◽  
Human Dermis ◽  
Subendothelial Matrix ◽  
Microvascular Endothelial ◽  
Endodermal Cells

The production of a basal lamina by microvascular endothelial cells (MEC) cultured on various substrata was examined. MEC were isolated from human dermis and plated on plastic dishes coated with fibronectin, or cell-free extracellular matrices elaborated by fibroblasts, smooth muscle cells, corneal endothelial cells, or PF HR9 endodermal cells. Examination of cultures by electron microscopy at selected intervals after plating revealed that on most substrates the MEC produced an extracellular matrix at the basal surface that was discontinuous, multilayered, and polymorphous. Immunocytochemical studies demonstrated that the MEC synthesize and deposit both type IV collagen and laminin into the subendothelial matrix. When cultured on matrices produced by the PF HR9 endodermal cells MEC deposit a subendothelial matrix that was present as a uniform sheet which usually exhibited lamina rara- and lamina densa-like regions. The results indicate that under the appropriate conditions, human MEC elaborate a basal lamina-like matrix that is ultrastructurally similar to basal lamina formed in vivo, which suggests that this experimental system may be a useful model for studies of basal lamina formation and metabolism.

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