Proteomic Profiling of Exosomes From Hemorrhagic Moyamoya Disease and Dysfunction of Mitochondria in Endothelial Cells

Stroke ◽  
2021 ◽  
Author(s):  
Xia Wang ◽  
Cong Han ◽  
Yangjie Jia ◽  
Jiayu Wang ◽  
Wei Ge ◽  
...  
Keyword(s):  
Mouse Brain ◽  
Moyamoya Disease ◽  
Molecular Mechanisms ◽  
Umbilical Vein ◽  
Neuroblastoma Cell ◽  
Human Hepatocyte ◽  
Control Group ◽  
Healthy Controls ◽  
Human Umbilical Vein ◽  
Ec Cells

Background and Purpose: Moyamoya disease (MMD) is a rare steno-occlusive and slowly progressing cerebrovascular disorder. The detailed mechanism of the underlying pathogenesis is still blurry. Methods: Tandem Mass Tag-labeled quantitative proteomics was performed on serum-derived exosomes (SDEs) extracted from adult patients diagnosed with pure ischemic MMD or hemorrhagic MMD and healthy controls. Then mouse brain vascular endothelial cell (EC), human umbilical vein EC, neuroblastoma cell, and human hepatocyte cell were treated with exosomes, and changes of the protein expression in mouse brain vascular EC cells were identified. Results: Proteomics analysis results showed that 859 shared proteins were detected in SDEs from ischemic and hemorrhagic MMD patients with 231 differently expressed compared with healthy controls. Bioinformatic analysis revealed dysregulated cell growth and maintenance and indicated disturbed actin dynamics in MMD, with CFL1 (Cofilin-1) and ACTR2/3 (actin-related protein 2/3; also known as ARP2/3) downregulated in ischemic and hemorrhagic patients’ SDEs. We also found immunity dysfunction in hemorrhagic MMD. Following treatment with MMD SDEs, mouse brain vascular EC cells showed significantly higher levels of proliferation and more ethynyl-2-deoxyuridine-positive cells compared with the healthy control group, while there were no obvious changes in the human umbilical vein EC and human hepatocyte cell. Interestingly, we also found that SDEs from ischemic MMD promoted neuroblastoma cell proliferation. Proteomic analysis of mouse brain vascular EC cells suggested that SDEs from hemorrhagic MMD patients induced dysfunction of the mitochondria in cerebrovascular ECs. Conclusions: This study highlighted potential molecular mechanisms underlying the pathogenesis of MMD patients, thereby providing new therapeutic strategies for MMD.

scholarly journals Hypoxic human umbilical vein endothelial cells induce activation of adherent polymorphonuclear leukocytes

Blood ◽  
1994 ◽  
Vol 83 (12) ◽  
pp. 3705-3716 ◽  
Author(s):  
T Arnould ◽  
C Michiels ◽  
J Remacle
Keyword(s):  
Endothelial Cells ◽  
Superoxide Dismutase ◽  
Superoxide Anion ◽  
Calcium Concentration ◽  
Molecular Mechanisms ◽  
Umbilical Vein ◽  
Superoxide Anion Production ◽  
Elastase Inhibitor ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells

Abstract Several pieces of evidence are reported for the accumulation of activated neutrophils in ischemic and reperfused tissues leading to the transformation of the ischemic tissue into an inflammatory territory and to an enhancement of tissue damages during reoxygenation. However, the molecular mechanisms responsible for these observations and the precise role played by endothelial cells in this process are still poorly understood. In this study, an in vitro model that mimics this situation was used to investigate the effects of hypoxia-incubated human umbilical vein endothelial cells (HUVEC) on polymorphonuclear leukocyte (PMN) functions. A strong PMN activation characterized by an increase in intracellular calcium concentration as well as by superoxide anion release and leukotriene B4 production was observed when these cells were coincubated with hypoxic HUVEC. On the other hand, conditioned medium from hypoxia-incubated HUVEC failed to activate PMN, as determined by the lack of PMN calcium concentration increase, the failure of superoxide anion production enhancement, as well as the absence of effects on the integrin CD18, CD11a, and CD11b expression. These results indicate that the presence of hypoxia- incubated HUVEC is necessary to obtain an activation of the PMN, probably via the adherence process. Once activated by coincubation with hypoxic HUVEC, PMN became cytotoxic, as evidenced by 51Cr released from prelabeled HUVEC. This cytotoxic effect of activated PMN for hypoxic endothelial cells could be prevented by a combination of superoxide dismutase and catalase (94% inhibition), whereas superoxide dismutase alone was inefficient. Antiprotease (alpha 2-macroglobulin) and a specific elastase inhibitor (MAAPV-CMK) were also inefficient. These results correlate very well with the fact that no increase in elastase release could be observed in supernatants from PMN coincubated with hypoxic HUVEC. Furthermore, when adherence process was blocked by oleic acid or by anti-ICAM-1 monoclonal antibodies, protection was, respectively, 90% and 72%. We thus evidenced that free radicals but not elastase released from activated PMN coincubated with hypoxic HUVEC are involved in HUVEC injury. We conclude from these results that PMN activation is initiated by PMN adherence to hypoxic HUVEC. These observations indicate that hypoxic HUVEC may be partly responsible for neutrophil activation observed in ischemic tissues, which is part of the amplification process of tissue damage.

scholarly journals Microcystins Induces Vascular Inflammation in Human Umbilical Vein Endothelial Cells via Activation of NF-κB

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Jun Shi ◽  
Jie Zhou ◽  
Min Zhang
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Adhesion Molecules ◽  
Cell Adhesion Molecules ◽  
Inflammatory Process ◽  
Molecular Mechanisms ◽  
Umbilical Vein ◽  
Health Hazard ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells

Microcystins (MCs) produced by toxic cyanobacteria cause serious water pollution and public health hazard to humans and animals. However, direct molecular mechanisms of MC-LR in vascular endothelial cells (ECs) have not been understood yet. In this study, we investigated whether MC-LR induces vascular inflammatory process in cultured human umbilical vein endothelial cells (HUVECs). Our data demonstrated that MC-LR decreased HUVECs proliferation and tube formation and enhanced apoptosis. MC-LR also induced intracellular reactive oxygen species formation (ROS) in HUVECs. The MC-LR directly stimulated phosphorylation of NF-κB. Furthermore, MC-LR also increased cell adhesion molecules (ICAM-1 and VCAM-1) expression in HUVECs. Taken together, the present data suggested that MC-LR induced vascular inflammatory process, which may be closely related to the oxidative stress, NF-κB activation, and cell adhesion molecules expression in HUVECs. Our findings may highlight that MC-LR causes potential damage to blood vessels.

scholarly journals The expression of Drosha, DGCR8, Dicer and Ago-2 genes are upregulated in human umbilical vein endothelial cells under hyperglycemic condition

2018 ◽  
Vol 52 (3) ◽  
pp. 123-127 ◽  
Author(s):  
Farhad Ghadiri Soufi ◽  
Ali Akbar Poursadegh Zonouzi ◽  
Ebrahim Eftekhar ◽  
Kamila Kamali ◽  
Sara Aghakhani Chegeni ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Mrna Expression ◽  
Umbilical Vein ◽  
Control Group ◽  
Human Umbilical Vein ◽  
Expression Levels ◽  
Expression Of Genes ◽  
Mirnas Expression ◽  
Umbilical Vein Endothelial Cells ◽  
Mrna Expression Levels

AbstractObjectives. It has been shown that dysregulation of miRNAs expression contributes to the pathogenesis and progression of the diabetes and diabetes-related complications. Drosha, DGCR8, Dicer, and Ago-2 are involved in the miRNA maturation. The aim of the present study was to investigate the mRNA expression levels of these genes in the human umbilical vein endothelial cells (HUVECs) under hyperglycemic condition.Methods. HUVECs were cultured in normo-(5 mM) and hyperglycemic (25 mM) conditions for 24 h. As osmotic control, cells were treated with D-mannitol (25 mM, for 24 h). The mRNA expression levels of Drosha, DGCR8, Dicer and Ago-2 were evaluated using quantitative real-time PCR.Results. The expression level of Drosha, DGCR8, Dicer, and Ago-2 were increased in hyperglycemic HUVECs compared to the control group.Conclusion. Our results show that under hyperglycemic condition, expression of genes involved in the miRNA maturation was significantly increased in HUVECs. Upregulation of these genes may have role in diabetic complications through the dysregulation of the miRNA expression.

Blood-brain barrier formation of grafted human umbilical vein endothelial cells in athymic mouse brain

2000 ◽  
Vol 858 (1) ◽  
pp. 172-176 ◽  
Author(s):  
Hideyuki Akiyama ◽  
Takeshi Kondoh ◽  
Takashi Kokunai ◽  
Tatsuya Nagashima ◽  
Naoaki Saito ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Blood Brain Barrier ◽  
Mouse Brain ◽  
Umbilical Vein ◽  
Brain Barrier ◽  
Athymic Mouse ◽  
Human Umbilical Vein ◽  
Blood Brain ◽  
Barrier Formation ◽  
Umbilical Vein Endothelial Cells

scholarly journals A Randomised, Controlled Study of Different Glycaemic Targets during Gestational Diabetes Treatment: Effect on the Level of Adipokines in Cord Blood and ANGPTL4 Expression in Human Umbilical Vein Endothelial Cells

2018 ◽  
Vol 2018 ◽  
pp. 1-8 ◽  
Author(s):  
P. Popova ◽  
L. Vasilyeva ◽  
A. Tkachuck ◽  
M. Puzanov ◽  
A. Golovkin ◽  
...  
Keyword(s):  
Gene Expression ◽  
Endothelial Cells ◽  
Gestational Diabetes ◽  
Cord Blood ◽  
Umbilical Vein ◽  
Fasting Blood Glucose ◽  
Controlled Study ◽  
Control Group ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells

Our aim was to study the expression of adipokine-encoding genes (leptin, adiponectin, and angiopoietin-like protein 4 (ANGPTL4)) in human umbilical vein endothelial cells (HUVECs) and adipokine concentration in cord blood from women with gestational diabetes mellitus (GDM) depending on glycaemic targets. GDM patients were randomised to 2 groups per target glycaemic levels: GDM1 (tight glycaemic targets, fasting blood glucose < 5.1 mmol/L and <7.0 mmol/L postprandial, N=20) and GDM2 (less tight glycaemic targets, <5.3 mmol/L and < 7.8 mmol/L, respectively, N=21). The control group included 25 women with normal glucose tolerance. ANGPTL4 expression was decreased in the HUVECs from GDM patients versus the control group (23.11 ± 5.71, 21.47 ± 5.64, and 98.33 ± 20.92, for GDM1, GDM2, and controls; p<0.001) with no difference between GDM1 and GDM2. The level of adiponectin gene expression was low and did not differ among the groups. Leptin gene expression was undetectable in HUVECs. In cord blood, leptin/adiponectin ratio (LAR) was increased in GDM2 compared to controls and GDM1 (p=0.038) and did not differ between GDM1 and controls. Tight glycaemic targets were associated with normalisation of increased LAR in the cord blood. ANGPTL4 expression was downregulated in HUVECs of newborns from GDM mothers and was not affected by the intensity of glycaemic control.

scholarly journals Sevoflurane inhibits Human umbilical vein endothelial cells migration by up-regulating VE-cadherin expression

2020 ◽  
Author(s):  
Yuanqing Sun ◽  
Zifeng Xu ◽  
Tao Xu ◽  
Hui Xu
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Control Group ◽  
Vascular Endothelial ◽  
Minimal Alveolar Concentration ◽  
Migration Distance ◽  
Inhalation Anesthesia ◽  
Human Umbilical Vein ◽  
Significant Difference ◽  
Umbilical Vein Endothelial Cells

Abstract Background Sevoflurane is a commonly used inhalation anesthesia and is famous for rapid onset of action, less metabolism in vivo and fast recovery. The aim of this study is to elaborate whether sevoflurane inhibits Human umbilical vein endothelial cells (HUVECs) migration function.Methods In vitro experiments, the HUVECs were divided into four groups randomly, and were exposed to 2% sevoflurane, refer to 1.6 minimal alveolar concentration (MAC), respectively for 0.5h, 1h, 2h and the first group was the control group which exposed to the same gas environment with other three groups but only without sevoflurane. After sevoflurane exposure, HUVECs were conducted the scratch assay.Results The results suggested that the HUVECs exposed to 2% sevoflurane for 2 h were more obviously inhibited on the migration distance during 12 h after scratched than the control group. Quantitative PCR results suggested that the HUVECs exposed to 2% sevoflurane for 2 h expressed more vascular endothelial cadherin (VE-cadherin) than the control group, with statistic difference. However, other scratch assay and quantitative trials suggested that the HUVECs which were transfected with VE-cadherin siRNA and exposed to 2% sevoflurane for 2 h had no significant difference with the control group on the migration distance and the expression of VE-cadherin.Conclusion These results suggested that sevoflurane inhibited the HUVECs migration function by up-regulating VE-cadherin expression, and may have an adverse effect on the normal functions of vascular endothelial cells.

scholarly journals miR-1273g-3p participates in acute glucose fluctuation-induced autophagy, dysfunction, and proliferation attenuation in human umbilical vein endothelial cells

2016 ◽  
Vol 310 (9) ◽  
pp. E734-E743 ◽  
Author(s):  
Jun Guo ◽  
Yaxiong Sang ◽  
Tao Yin ◽  
Bo Wang ◽  
Wenping Yang ◽  
...  
Keyword(s):  
Wound Healing ◽  
Endothelial Cells ◽  
Endothelial Dysfunction ◽  
Critically Ill ◽  
High Mortality ◽  
Critically Ill Patients ◽  
Umbilical Vein ◽  
Control Group ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells

Acute glucose fluctuations (AGF) often cause high mortality among critically ill patients, but the mechanisms induced by AGF are not clear. Recent studies suggest that endothelial dysfunction is a key factor that leads to high mortality among critically ill patients. Our goal is to evaluate the phenomenon and mechanisms of endothelial dysfunction induced by AGF. In this study, the functions of human umbilical vein endothelial cells (HUVECs) were compared after treatment with sustained high glucose (SHG), AGF in two groups (AGF1 fluctuations between 5 and 16 mM and AGF2 fluctuations between 5 and 25 mM), and normal glucose levels as a control group (CTR). The medium of the groups was changed every 4 h. The influence of AGF on wound healing was also tested on C57BL/6 mice. The results show that cell proliferation, angiogenesis, and migration functions were injured in the SHG and both AGF groups. AGF2 group shows the worse condition in vitro. In vivo, the wound healing was delayed after the AGF treatment. Furthermore, the markers of apoptosis and autophagy were analyzed. We observed that the autophagy changed in all treatment groups, but apoptosis showed no change. To get to know the mechanism of dysfunction and autophagy, we performed the microRNA chip assay and real-time PCR and found miR-1273g-3p remarkably changed in AGF2 group. After the mimic and inhibitor of miR-1273g-3p were transfected during the AGF2 treatment, we found that the dysfunction and autophagy were partially enhanced by miR-1273g-3p mimic and reversed by miR-1273g-3p inhibitor in AGF2 group. Thus, we conclude that AGF can induce more dysfunction and autophagy, and miR-1273g-3p is also an important factor that leads to the injury.

scholarly journals Extracellular HtrA2 Induces Apoptosis in Human Umbilical Vein Endothelial Cells

2019 ◽  
Vol 20 (21) ◽  
pp. 5446
Author(s):  
Kaur ◽  
Stallmann ◽  
Schanze ◽  
Laumann ◽  
Heger ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Molecular Mechanisms ◽  
Umbilical Vein ◽  
Ischemia Reperfusion ◽  
Annexin V ◽  
Myocardial Damage ◽  
Human Umbilical Vein ◽  
St Segment Elevation ◽  
Induced Apoptosis ◽  
Umbilical Vein Endothelial Cells

The serine protease high-temperature-required protein A2 (HtrA2) has been identified as a key intracellular molecule promoting apoptosis in cells during ischemia reperfusion (IR) injury. IR injury in ST-segment elevation myocardial infarction (STEMI) contributes to overall myocardial damage. HtrA2 has further been shown to be significantly increased in the serum of patients with STEMI. In the present pilot study, we use human umbilical vein endothelial cells (HUVECs) to investigate whether extracellular HtrA2 induces apoptosis using Annexin V staining. Furthermore, we examine whether HtrA2 is released extracellularly after staurosporine-induced apoptosis using ELISA. We find that HtrA2 is released upon induction of apoptosis by staurosporine into the cell culture medium. Furthermore, treatment of HUVECs with extracellular HtrA2-induces apoptosis, while the addition of anti-HtrA2 antibodies reduces both HtrA2- and staurosporine-induced endothelial cell apoptosis. In conclusion, we show here that extracellular HtrA2 induces apoptosis in human endothelial cells, although the exact molecular mechanisms have to be investigated in future.

scholarly journals PPAR-αAgonist Fenofibrate Upregulates Tetrahydrobiopterin Level through Increasing the Expression of Guanosine 5′-Triphosphate Cyclohydrolase-I in Human Umbilical Vein Endothelial Cells

PPAR Research ◽  
2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Jinbo Liu ◽  
Changlin Lu ◽  
Fuwang Li ◽  
Haining Wang ◽  
Liyun He ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Control Group ◽  
Peroxisome Proliferator ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Peroxisome Proliferator Activated Receptor ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells

Tetrahydrobiopterin (BH4) is an essential cofactor for endothelial nitric oxide (NO) synthase. Guanosine 5′-triphosphate cyclohydrolase-I (GTPCH-I) is a key limiting enzyme for BH4 synthesis. In the present in vitro study, we investigated whether peroxisome proliferator-activated receptorα(PPAR-α) agonist fenofibrate could recouple eNOS by reversing low-expression of intracellular BH4 in endothelial cells and discussed the potential mechanisms. After human umbilical vein endothelial cells (HUVECs) were treated with lipopolysaccharide (LPS) for 24 hours, the levels of cellular eNOS, BH4 and cell supernatant NO were significantly reduced compared to control group. And the fluorescence intensity of intracellular ROS was significantly increased. But pretreated with fenofibrate (10 umol/L) for 2 hours before cells were induced by LPS, the levels of eNOS, NO, and BH4 were significantly raised compared to LPS treatment alone. ROS production was markedly reduced in fenofibrate group than LPS group. In addition, our results showed that the level of intracellular GTPCH-I detected by western blot was increased in a concentration-dependent manner after being treated with fenofibrate. These results suggested that fenofibrate might help protect endothelial function and against atherosclerosis by increasing level of BH4 and decreasing production of ROS through upregulating the level of intracellular GTPCH-I.

Effect of Chlorogenic Acid on H2O2-Induced Dysfunction and Apoptosis in HUVECs

2013 ◽  
Vol 706-708 ◽  
pp. 69-72 ◽  
Author(s):  
Ping Li ◽  
Luo Sha Zhao ◽  
Yin Man Feng ◽  
Xian Li Yuan ◽  
Shu Lian Liu ◽  
...  
Keyword(s):  
Chlorogenic Acid ◽  
Mrna Expression ◽  
Molecular Mechanisms ◽  
Fluorescence Probe ◽  
Umbilical Vein ◽  
Control Group ◽  
Dependent Manner ◽  
Protective Effects ◽  
Acid Pretreatment ◽  
Enos Mrna

To investigate the protective effects and the possible molecular mechanisms of chlorogenic acid on H2O2-induced dysfunction and apoptosis in human umbilical vein endothelial cells (HUVECs). HUVECs were divided into three groups: H2O2groups (0, 5, 10, 20, 50mM), chlorogenic acid (100nmol/L) groups and atorvastatin (10μmol/L) groups. The eNOS mRNA expression was measured by RT-PCR. The content of NO in the supernatant was detected by the nitrate reductase assay. Cells activity was evaluated by the MTT assay. The level of reactive oxygen species (ROS) in cells was measured using H2DCF-DA as a fluorescence probe. Results demonstrated that H2O2caused a significant down-regulation of eNOS mRNA expression and decreased the secretion of NO in a dose-dependent manner compared with the control group. Furthermore, H2O2 induced cell apoptosis, increased ROS accumulation. Nevertheless, these effects of H2O2could be blocked by chlorogenic acid in chlorogenic acid pretreatment groups. CONCLUSION:chlorogenic acid can inhibite H2O2-induced dysfunction and apoptosis in HUVECs, which may be related to its effects on suppressing oxidative stress, up-regulating eNOS pathway. These results suggest the usefulness of chlorogenic acid particularly in the treatment of cardiovascular disease.

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