Endothelial Cell Injury Under High Frequency Vibration in the Rat-Tail Model

2011 ◽  
Author(s):  
Shilpi Goenka ◽  
Srikara V. Peelukhana ◽  
Jay Kim ◽  
Keith F. Stringer ◽  
Rupak K. Banerjee
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Oxidative Damage ◽  
Cell Injury ◽  
Industrial Workers ◽  
Injury Model ◽  
Blue Stain ◽  
Hematoxylin And Eosin ◽  
Rat Tail Model ◽  
Rat Tail

Hand-Arm Vibration Syndrome (HAVS) consists of vascular, sensorineural and musculoskeletal disorders and affects around 1.7–5.8% of industrial workers. In this study, a rat-tail vibration injury model is used to assess early vascular damage due to HAVS, manifested in the form of endothelial cell vacuolation and oxidative injury. Tails were vibrated at two frequencies 125Hz and 250Hz for 4hr/day for 1 and 5 days (49m/s2). Hematoxylin and Eosin (H&E) staining was done to assess gross changes in artery sections and toluidine blue stain was done for vacuole counting. Immunohistochemical (IHC) methods were used to detect Nitrotyrosine, a potent biomarker of cell inflammation and oxidative stress. The vacuole count in Endothelial Cells (ECs) was not statistically significant after 1 and 5 days for any frequency. However IHC images showed significant oxidative damage in Endothelial Cells (ECs) with considerable oxidative damage being induced as early as 1 day for both 125Hz and 250Hz frequencies, with more EC damage induced by 250Hz frequency after 5 days. These findings indicate that higher frequency vibrations can cause severe oxidative damage to EC.

Peptide5 attenuates rtPA related brain microvascular endothelial cells reperfusion injury via the Wnt/β-catenin signalling pathway

2021 ◽  
Vol 18 ◽  
Author(s):  
Weimin Ren ◽  
Chuyi Huang ◽  
Heling Chu ◽  
Yuping Tang ◽  
Xiaobo Yang
Keyword(s):  
Endothelial Cells ◽  
Ischemic Stroke ◽  
Endothelial Cell ◽  
Cell Injury ◽  
Time Window ◽  
Microvascular Endothelial Cells ◽  
Brain Microvascular Endothelial Cells ◽  
Endothelial Cell Injury ◽  
Endothelial Cell Death ◽  
Therapeutic Time Window

Aims: Brain vascular endothelial cell dysfunction after rtPA treatment is a significant factor associated with poor prognosis, suggesting that alleviation of rtPA-related endothelial cell injury may represent a potential beneficial strategy along with rtPA thrombolysis. Background: Thrombolysis with recombinant tissue plasminogen activator (rtPA) is beneficial for acute ischemic stroke but may increase the risk of hemorrhagic transformation (HT), which is considered ischemia-reperfusion injury. The underlying reason may contribute to brain endothelial injury and dysfunction related to rtPA against ischemic stroke. As previous studies have demonstrated that transiently blocked Cx43 using peptide5 (Cx43 mimetic peptide) during retinal ischemia reduced vascular leakage, it is necessary to know whether this might help decrease side effect of rtPA within the therapeutic time window. Objective: This study aims to investigate the effects of peptide5 on rtPA-related cell injury during hypoxia/reoxygenation (H/R) within the therapeutic time window. Methods: In this study, we established a cell hypoxia/reoxygenation H/R model in cultured primary rat brain microvascular endothelial cells (RBMECs) and evaluated endothelial cell death and permeability after rtPA treatment with or without transient peptide5. In addition, we also investigated the potential signaling pathway to explore the underlying mechanisms preliminarily. Results: The results showed that peptide5 inhibited rtPA-related endothelial cell death and permeability. It also slightly increased tight junction (ZO-1, occluding, claudin-5) and β-catenin mRNA expression, demonstrating that peptide5 might attenuate endothelial cell injury by regulating the Wnt/β-catenin pathway. The following bioinformatic exploration from the GEO dataset GSE37239 was also consistent with our findings. Conclusion: This study showed that the application of peptide5 maintained cell viability and permeability associated with rtPA treatment, revealing a possible pathway that could be exploited to limit rtPA-related endothelial cell injury during ischemic stroke. Furthermore, the altered Wnt/β-catenin signaling pathway demonstrated that signaling pathways associated with Cx43 might have potential applications in the future. This study may provide a new way to attenuate HT and assist the application of rtPA in ischemic stroke.

scholarly journals Omentin-1 Ameliorated Free Fatty Acid-Induced Impairment in Proliferation, Migration, and Inflammatory States of HUVECs

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Yubin Chen ◽  
Fen Liu ◽  
Fei Han ◽  
Lizhi Lv ◽  
Can-e Tang ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Fatty Acid ◽  
Endothelial Cell ◽  
Free Fatty Acid ◽  
Cell Injury ◽  
Umbilical Vein ◽  
Quantitative Polymerase Chain Reaction ◽  
Cell Counting ◽  
Endothelial Cell Injury ◽  
Umbilical Vein Endothelial Cells

Objectives. Endothelial cell injury is a critical pathological change during the development of atherosclerosis. Here, we explored the effect of omentin-1 on free fatty acid- (FFA-) induced endothelial cell injury. Methods. An FFA-induced endothelial cell injury model was established to investigate the role of omentin-1 in this process. Cell proliferation was analyzed with the Cell Counting Kit assay and flow cytometry. Scratch and transwell assays were used to evaluate cell migration. Factors secreted by endothelial cells after injury were detected by western blotting, reverse-transcription quantitative polymerase chain reaction, and cellular fluorescence assay. Results. Omentin-1 rescued the FFA-induced impaired proliferation and migration capabilities of human umbilical vein endothelial cells (HUVECs). It decreased the number of THP-1 cells attached to HUVECs in response to injury and inhibited the FFA-induced proinflammatory state of HUVECs. Conclusion. Omentin-1 could partly ameliorate FFA-induced endothelial cell injury.

scholarly journals Human Endothelial Cell Injury Mechanisms in Vitro

1977 ◽  
Author(s):  
R.T. Wall ◽  
L.A. Harker ◽  
G. Striker ◽  
L. Quadracci
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Dose Response ◽  
Cell Injury ◽  
Cultured Cells ◽  
Test Material ◽  
51Cr Release ◽  
Injury Mechanisms ◽  
Cell Antibody

Since endothelial cell desquamation constitutes the initial event leading to acute and chronic vascular diseases, we have developed an in vitro system for studying injury mechanisms using cultured cells. The system involves the incubation of test material with confluent, 51Cr-labeled, cultured human umbilical vein endothelial cells in 100% pooled human serum; injury was measured as% endothelial cell 51Cr release (ECR) into the supernatant media. Baseline spontaneous ECR was 6.0% ± 1.5 while specific rabbit antihuman endothelial cell antibody (1:64 dilution) in the presence of fresh complement released 90% ± 3 of the total releasable 51Cr activity. Consistent with in vivo predictions, homocysteine in concentrations of 0.5-40 mM iduced dose response ECR up to 20%. Neither methionine nor homocystine increased ECR over controls. Also as expected from experimental work endotoxin (S. enteritidis) caused dose response ECR, i.e., 24% ± 2 at 10 ug/ml. Control studies with cultured human smooth muscle cells (SMC) demonstrated no ECR with homocysteine but significant release occurred with endotoxin.Specific complement dependent cytotoxic anti-endothelial cell antibody was demonstrated in the secum of two thrombotic thrombocytopenic purpura (TTP) patients at a 1:1 dilution, inducing ECR of 23.1% ± 1.4 and 21.0% ± 0.25. The antibody was also demonstrated by immunoflourescent techniques and was absorbed from the serum using human endothelial cells. One disease-free, six month survivor showed no cytotoxic activity. Serum from a patient with adult hemolytic-uremic syndrome demonstrated antibody dependent cell mediated cytotoxicity with release of 22% ± 1.1 when normal non-immune lymphocytes were added to heat inactivated seriun. Control studies with SMC showed no ECR with TTP sera. We conclude that assays of endothelial cell 51Cr release are stable, reproducible and useful in the characterization of injury mechanisms.

scholarly journals Acacetin improves endothelial dysfunction and aortic fibrosis in insulin-resistant SHR rats by estrogen receptors

2020 ◽  
Vol 47 (9) ◽  
pp. 6899-6918
Author(s):  
Yaxin Wei ◽  
Peipei Yuan ◽  
Qi Zhang ◽  
Yang Fu ◽  
Ying Hou ◽  
...  
Keyword(s):  
Estrogen Receptor ◽  
Endothelial Cell ◽  
Endothelial Dysfunction ◽  
Estrogen Receptors ◽  
Cell Injury ◽  
Shr Rats ◽  
Endothelial Cell Injury ◽  
Insulin Resistant ◽  
Injury Model ◽  
Huvec Cells

Abstract The aim of the work was to investigate the effects of acacetin on endothelial dysfunction and aortic fibrosis in insulin-resistant SHR rats and explore its mechanism. Seven-week-old male spontaneously hypertensive rats (SHR) were selected to establish a rat model of hypertension with insulin resistance induced by 10% fructose. The nuclear factor kappa B p65 (NF-κB p65) and Collagen I were observed by Immunohistochemistry. Immunofluorescence was used to observe estrogen receptor-alpha (ERα), estrogen receptor-beta (ERβ), and G protein-coupled receptor 30 (GPR30). Western blotting was used to detect interleukin (IL-1β), Arginase 2 (ARG2), Nostrin, endothelial nitric oxide synthase (eNOS), TGF-β, Smad3, ERK pathway proteins such as p-c-Raf, p-MEK1/2, p-ERK, ERK, p-P90RSK and p-MSK1. We found that acacetin did have an improvement on endothelial dysfunction and fibrosis. Meanwhile, it was also found to have a significant effect on the level of estrogen in this model by accident. Then, the experiment of uterine weight gain in mice confirmed that acacetin had a certain estrogen-like effect in vivo and played its role through the estrogen receptors pathway. In vitro experience HUVEC cells were stimulated with 30 mM/L glucose and 100 mM/L NaCl for 24 h to establish the endothelial cell injury model. HUVEC cells were treated with 1 μM/L estrogen receptors antagonist (ICI 182780) for 30 min before administration. Cell experiments showed that acacetin could reduce the apoptosis of HUVEC cells, the levels of inflammatory cytokines and the expression of TGF-β, Collagen I and Smad3 in endothelial cell injury model. After treatment with ICI 182780, the improvement of acacetin was significantly reversed. The results showed that acacetin relieved endothelial dysfunction and reduced the aortic fibrosis in insulin-resistant SHR rats by reducing the release of inflammatory factors and improving vasodilatory function through estrogen signaling pathway.

scholarly journals Human Cytomegalovirus-Specific CD4+-T-Cell Cytokine Response Induces Fractalkine in Endothelial Cells

2004 ◽  
Vol 78 (23) ◽  
pp. 13173-13181 ◽  
Author(s):  
Cynthia A. Bolovan-Fritts ◽  
Rodney N. Trout ◽  
Stephen A. Spector
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
T Cell ◽  
Mononuclear Cells ◽  
Cell Injury ◽  
Transplant Rejection ◽  
Vascular Diseases ◽  
T Cell Response ◽  
Peripheral Blood Mononuclear ◽  
Necrosis Factor Alpha

ABSTRACT Cytomegalovirus (CMV) infection has been linked to inflammation-related disease processes in the human host, including vascular diseases and chronic transplant rejection. The mechanisms through which CMV affects the pathogenesis of these diseases are for the most part unknown. To study the contributing role of the host immune response to CMV in these chronic inflammatory processes, we examined endothelial cell interactions with peripheral blood mononuclear cells (PBMC). Endothelial cultures were monitored for levels of fractalkine induction as a marker for initiating the host inflammatory response. Our results demonstrate that in the presence of CMV antigen PBMC from normal healthy CMV-seropositive donors produce soluble factors that induce fractalkine in endothelial cells. This was not observed in parallel assays with PBMC from seronegative donors. Examination of subset populations within the PBMC further revealed that CMV antigen-stimulated CD4+ T cells were the source of the factors, gamma interferon and tumor necrosis factor alpha, driving fractalkine induction. Direct contact between CD4+ cells and the endothelial monolayers is required for this fractalkine induction, where the endothelial cells appear to provide antigen presentation functions. These findings indicate that CMV may represent one member of a class of persistent pathogens where the antigen-specific T-cell response can result in the induction of fractalkine, leading to chronic inflammation and endothelial cell injury.

scholarly journals Atherosclerosis-Associated Endothelial Cell Apoptosis by MiR-429-Mediated Down Regulation of Bcl-2

2015 ◽  
Vol 37 (4) ◽  
pp. 1421-1430 ◽  
Author(s):  
Tao Zhang ◽  
Feng Tian ◽  
Jing Wang ◽  
Jing Jing ◽  
Shan-Shan Zhou ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Cell Apoptosis ◽  
Cell Injury ◽  
Luciferase Reporter ◽  
Lower Percentage ◽  
Endothelial Cell Apoptosis ◽  
Down Regulation ◽  
Bioinformatics Analyses

Background/Aims: Endothelial cell injury and subsequent apoptosis play a key role in the development and pathogenesis of atherosclerosis, which is hallmarked by dysregulated lipid homeostasis, aberrant immunity and inflammation, and plaque-instability-associated coronary occlusion. Nevertheless, our understanding of the mechanisms underlying endothelial cell apoptosis is still limited. MicroRNA-429 (miR-29) is a known cancer suppressor that promotes cancer cell apoptosis. However, it is unknown whether miR-429 may be involved in the development of atherosclerosis through similar mechanisms. We addressed these questions in the current study. Methods: We examined the levels of endothelial cell apoptosis in ApoE (-/-) mice suppled with high-fat diet (HFD), a mouse model for atherosclerosis (simplified as HFD mice). We analyzed the levels of anti-apoptotic protein Bcl-2 and the levels of miR-429 in the purified CD31+ endothelial cells from mouse aorta. Prediction of the binding between miR-429 and 3'-UTR of Bcl-2 mRNA was performed by bioinformatics analyses and confirmed by a dual luciferase reporter assay. The effects of miR-429 were further analyzed in an in vitro model using oxidized low-density lipoprotein (ox-LDL)-treated human aortic endothelial cells (HAECs). Results: HFD mice developed atherosclerosis in 12 weeks, while the control ApoE (-/-) mice that had received normal diet (simplified as NOR mice) did not. HFD mice had significantly lower percentage of endothelial cells and significantly higher percentage of mesenchymal cells in the aorta than NOR mice. Significantly higher levels of endothelial cell apoptosis were detected in HFD mice, resulting from decreases in Bcl-2 protein, but not mRNA. The decreases in Bcl-2 in endothelial cells were due to increased levels of miR-429, which suppressed the translation of Bcl-2 mRNA via 3'-UTR binding. These in vivo findings were reproduced in vitro on ox-LDL-treated HAECs. Conclusion: Atherosclerosis-associated endothelial cell apoptosis may result from down regulation of Bcl-2, through increased miR-429 that binds and suppresses translation of Bcl-2 mRNA.

Dependence of Higher Frequency Components on Bone Tissue Alterations in the Rat-Tail Model

2013 ◽  
Author(s):  
Brian Kim ◽  
Shilpi Goenka ◽  
Srikara V. Peelukhana ◽  
Keith F. Stringer ◽  
Jay H. Kim ◽  
...  
Keyword(s):  
Structural Damage ◽  
Statistical Significance ◽  
Excitation Frequency ◽  
Control Group ◽  
Vibration Experiment ◽  
Frequency Components ◽  
Hematoxylin And Eosin ◽  
Rat Tail Model ◽  
Tissue Alterations ◽  
Rat Tail

Through a recently conducted rat-tail vibration experiment, we have been able to determine that the tested frequencies of vibration have a significant effect on biochemical damage signified by nitro-tyrosine (NT) staining on trabecular bone, while structural damage quantified through a Hematoxylin and Eosin (H&E) stain on cortical bone exhibited statistical significance only for the 250 Hz group compared to the control group. These results seem to indicate a relationship between the growing quantities of biochemical damage when increasing the excitation frequency, thus further experimentation for frequencies between 250 Hz and 400 Hz is recommended.

IMMUNOLOGICAL AND CHEMICAL DETECTION OF VAC IN CULTURED ENDOTHELIAL CELLS

1987 ◽  
Author(s):  
C P M Reutelingsperger ◽  
W Buurman ◽  
G Horn-stra ◽  
H C Horn-stra
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Cell Injury ◽  
Polyclonal Antibodies ◽  
Human Umbilical Cord ◽  
Nuclear Area ◽  
Intracellular Protein ◽  
Coagulation Assay ◽  
Chemical Techniques ◽  
Extracellular Side

Recently we reported the presence in human umbilical cord vessels of an anticoagulatory protein (VAC, Mr = 32,000) which inhibits phospholipid dependent procoagulant reactions through a high affinity binding, in the presence of calcium, to the phospholipid surface. The mechanism of anticoagulation differs fundamentally from those of the well-known physiological anticoagulants.Polyclonal antibodies, raised in rabbits against purified VAC, bind in cultured endothelial cell lysates to an antigen with Mr = 32,000, as was revealed with immunoblotting techniques. It is demonstrated with chemical techniques, that this antigen is identical to VAC.VAC appears to be an intracellular protein, attached to fine granular structures, which are located outside the nuclear area. Quiescent endothelial cells do neither secrete VAC in detectable amounts, nor contain detectable VAC on the extracellular side of their plasma membrane. In the presence of 1 mM calcium, endothelial VAC binds reversibly, as was indicated with EDTA, to the subcellular structures of the endothelial cell. Once VAC is bound to the subcellular structures, their apparent procoagulant activities are diminished, as was shown in a one-stage coagulation assay.Based on these findings, we propose that the presence of VAC in endothelial cells supplies the endothelium with an additional anticoagulatory mechanism, which can be activated after cell injury, when intracellular structures become exposed to plasma constituents. VAC then controls the formation of procoagulant complexes, localized to the subcellular structures.

Activation of endothelial cells, coagulation and fibrinolysis in children with Dengue virus infection

2007 ◽  
Vol 97 (04) ◽  
pp. 627-634 ◽  
Author(s):  
Panya Seksarn ◽  
Sureeporn Pongsewalak ◽  
Usa Thisyakorn ◽  
Jeanne Lusher ◽  
Darintr Sosothikul
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Dengue Virus ◽  
Plasminogen Activator ◽  
Cell Activation ◽  
Cell Injury ◽  
Clinical Severity ◽  
Coagulation Cascade ◽  
Severe Bleeding ◽  
Pai 1

SummaryDengue virus causes a febrile illness: Dengue fever (DF), and less frequently a life-threatening illness: Dengue hemorrhagic fever (DHF). Although severe bleeding remains a major cause of death in DHF, the pathogenesis of bleeding is poorly understood. This prospective cohort study was designed to determine the extent of activation of endothelial cells and the hemostatic system in correlation with clinical severity, and also to detect the best prognostic factor(s) for DHF. Endothelial cell activation, coagulation, anticoagulant and fibrinolysis parameters were measured in 42 children with Dengue infections (20 with DF and 22 with DHF) during three phases of illness. In DHF patients, during the febrile phase, vonWillebrand factor antigen (vWF:Ag),tissue factor (TF) and plasminogen activator inhibitor (PAI-1) were significantly elevated, while platelet counts andADAMTS 13 (a disintegrin and metalloprotease with thrombospondin repeats) were significantly low compared to DF patients. During the toxic phase, soluble thrombomodulin (sTM), tissue plasminogen activator (t-PA) and PAI-1 were also significantly increased, while ADAMTS 13 and thrombin activatable fibrinolysis inhibitor (TAFIa) were significantly low compared to DF patients. Abnormal vWF multimers were seen only in DHF patients. For endothelial cell injury and release of procoagulant components, activation of the coagulation cascade with thrombin generation, increased antifibrinolytic factors and consumption of natural anticoagulants, each appeared to play an important role in the development of hemorrhage in Dengue patients. Using logistic regression analysis, we found plasma VWF:Ag to be the best indicator of progression to DHF.

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