Expression Levels Of Vascular Cell Adhesion Molecule-1 Research Articles

IL-33 Suppresses the Progression of Atherosclerosis via the ERK1/2-IRF1-VCAM-1 Pathway.

This study was designed to explore the effects of interleukin 33 (IL-33) on the progression of atherosclerosis and the possible mechanism. The adhesion assay was performed on isolated peripheral blood mononuclear cells (PBMCs) and human umbilical vein endothelial cells (HUVEC). The expression of proteins and messenger RNA (mRNA) were detected by western blot and quantitative real-time polymerase chain reaction (PCR), including intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and P-selectin. The effect of IL-33 on the interaction of growth stimulation expressed gene 2 (ST2) with myeloid differentiation factor 88 (MyD88) and interleukin-1 receptor-associated kinase (IRAK) 1/4 were investigated using co-immunoprecipitation assay. An apolipoprotein (Apo) E-/- mice model was used to confirm the effect of IL-33 on atherosclerosis progression. Area of plaques was recorded by hematoxylin-eosin (H&E) staining. The severity of atherosclerosis plaque was evaluated using immunohistochemistry assay, and lipid accumulation was measured by an oil red O staining. In contrast, western blot was performed to detect the expression levels of VCAM-1, extracellular signal-regulated kinase (ERK) 1/2, and interferon regulatory factor 1 (IRF1). Our study observed that IL-33 suppressed cell adhesion and the expression of VCAM-1 in tumor necrosis factor-α (TNF-α) exposed HUVEC. Moreover, the addition of IL-33 significantly inhibited the expression of IRF1 and the binding level of IRF1 to VCAM-1 and also promoted the phosphorylation level of IRAK1/4 and ERK1/2 compared to TNF-α-stimulated HUVEC. The ST2 neutralizing antibody or ERK pathway inhibitor SCH772984 reversed the regulatory effects of IL-33 on HUVEC, suggesting that IL-33 suppressed IRF1 and VCAM-1 dependent on binding to ST2 and activating the ERK1/2 signaling pathway. Further investigation in vivo confirmed that IL-33 decreased the expressions of IRF1 and VCAM-1 by activating the phosphorylation of ERK1/2 in the thoracic aorta of Apo E-/- mice. In conclusion, our results demonstrated that IL-33 plays a protective role in the progression of atherosclerosis by inhibiting cell adhesion via the ERK1/2-IRF1-VCAM-1 pathway. This study may provide a potential therapeutic way to prevent the development of atherosclerosis.

Aquaporin-8 overexpression is involved in vascular structure and function changes in placentas of gestational diabetes mellitus patients.

To study the role and mechanism of aquaporin-8 (AQP8) in placental vascular development in gestational diabetes mellitus (GDM), hematoxylin-eosin staining and immunohistochemistry were utilized to analyze the histopathological changes in placentas in GDM patients. Transwell, CCK-8, and tube formation assays were performed to examine cell migration, proliferation, and tube formation. AQP8, vascular cell adhesion molecule 1 (VCAM-1), tumor necrosis factor alpha (TNF)-α, and vascular endothelial growth factor (VEGF)-A expression levels were investigated. Relative to the control group, the placentas in the GDM group showed morphological changes, the number of microvessels in the placental villi arterioles was significantly higher, and the area of microvessels in the arterioles of placental villi was significantly lower. The expression levels of VCAM-1, TNF-α, VEGF-A, and AQP8 in the GDM placentas and human umbilical vein endothelial cells (HUVECs) stimulated by high glucose were significantly higher than those in the control group, and AQP8 was located in placental endothelial cells. Overexpression of glucose and AQP8 inhibited tube formation, migration, and proliferation in HUVECs. High glucose levels can induce dysfunction in vascular endothelial cells and lead to pathological changes in the placental vascular structure in GDM. AQP8 overexpression in placental GDM can inhibit endothelial cell behavior, cause endothelial cell dysfunction, and further participate in the occurrence and development of GDM placental vascular lesions.

Antcin K inhibits VCAM-1-dependent monocyte adhesion in human rheumatoid arthritis synovial fibroblasts.

BackgroundAntcin K, an extract of Antrodia cinnamomea (a medicinal mushroom endemic to Taiwan commonly used in Chinese medicine preparations), inhibits proinflammatory cytokine production and angiogenesis in human rheumatoid arthritis synovial fibroblasts (RASFs), major players in RA disease. Antcin K also inhibits disease activity in mice with collagen-induced arthritis (CIA). Up until now, the effects of Antcin K upon cell adhesion molecules (CAMs) were unknown.MethodsRA and healthy synovial tissue samples (n = 10 in each group) were retrieved from the Gene Expression Omnibus (GEO) database (accession code: GDS5401) to compare CAM and monocyte marker expressions. In addition, synovial tissue samples from six RA patients and six patients undergoing arthroscopy for trauma/joint derangement (healthy controls) were subjected to immunohistochemical (IHC) analysis. mRNA and protein expression levels were analyzed in RASFs using RT-qPCR (Reverse transcription-quantitative polymerase chain reaction) and Western blot. RASFs were incubated with Antcin K and examined for monocyte adherence by fluorescence microscopy. Ankle joint tissue specimens from a CIA mouse model and healthy controls were stained with hematoxylin and eosin (H&E) and Safranin-O/Fast Green to examine histological changes and evidence of bone loss. IHC analysis determined levels of vascular cell adhesion molecule 1 (VCAM-1) and CD11b in CIA ankle tissue and clinical synovial tissue.ResultsLevels of VCAM-1 expression were higher in the GEO database specimens and the study’s clinical samples of RA synovial tissue compared with the healthy specimens. Antcin K dose-dependently inhibited VCAM-1 expression and monocyte adhesion in RASFs. Antcin K also significantly inhibited levels of VCAM-1 and monocyte CD11b expression in CIA tissue. These effects appeared to be mediated by MEK1/2-ERK, p38, and AP-1 signaling.ConclusionsAntcin K seems promising for the treatment of RA and deserves further investigations.

SARS‐CoV‐2 Spike Protein Enhances Toxicant‐Induced Blood‐Brain Barrier Dysfunction

Exposure to toxicants such as benzo(a)pyrene (BaP), 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin (TCDD), or perfluorooctane sulfonate (PFOS), or viruses such as severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) is associated with neuropathology and blood‐brain barrier (BBB) dysfunction. Recently, toxicant exposure has been reported to result in poorer health outcomes of (SARS‐CoV‐2) infection. Thus, we hypothesized that co‐exposure to toxicants and SARS‐CoV‐2 will cause greater BBB disruption than exposure to toxicants or SARS‐CoV‐2 alone.BBB permeability was assessed in vitro by measuring transendothelial electrical resistance (TEER; ohms cm2) of a monolayer of primary adult human brain microvascular endothelial cells (hBMECs). Cells were allowed to form a confluent monolayer on a 96‐well plate outfitted with circuit electrodes. Cells were treated with BaP (2, 10, and 50 uM), TCDD (1.5, 15, and 150 nM), PFOS (3, 30, and 300 uM), or recombinant SARS‐CoV‐2 spike protein (SP; 0.1, 1, and 10 nM). In experiments to test the synergistic effects of toxicants and SP, cells were treated with toxicants, then immediately were treated with SP (10 nM). Cell viability was assessed 24 hours after treatment using Cell Proliferation Kit I (Roche Diagnostics, Inc). Gene expression of tight junction protein 1 (TJP1) and vascular cell adhesion molecule‐1 (VCAM‐1) was measured by RT‐qPCR. One‐ and two‐way ANOVA followed by Holm‐Sidak post‐hoc comparisons were performed to detect differences in TEER (resistance, ohms cm2) and cell viability (absorbance, nm). T‐tests were performed to compare RT‐qPCR data. Statistical significance was reached at p < 0.05. Technical triplicates were averaged on 4 different days to obtain N = 4/group.Recombinant SARS‐CoV‐2 spike protein did not significantly affect the resistance or leakiness of the monolayer of hBMECs at any of the concentrations tested. However, when hBMECs were treated with SP (10 nM) in conjunction with BaP or PFOS, TEER was significantly reduced beyond treatment with the toxicants alone. When compared to BaP treatment alone, SP significantly lowered TEER in hBMECs treated with 10 uM or 50 uM of BaP at 9 to 24 hours after treatment (Holm‐Sidak, t’s > 2.507, p’s < 0.04), indicating a synergistic effect of SP and BaP in increasing leakiness of the hBMEC barrier. Similarly, SP caused a significant reduction in TEER of hBMECs treated with 3 uM PFOS starting at 15 hours post treatment when compared to PFOS treatment alone (Holm‐Sidak, t’s > 2.273, p’s < 0.04). TEER of TCDD‐treated hBMECs were not affected by SP treatment. At 24 hours after treatment, cell viability and expression of VCAM‐1 and TJP1 were not altered by treatment by SP alone, nor did SP affect cell viability or gene expression of toxicant‐treated hBMECs. These results suggest that disruption of the hBMEC monolayer was not due to cell death, or expression levels of VCAM‐1 and TJP1.To our knowledge, this is the first report of the synergistic effects of SARS‐CoV‐2 spike protein and environmental toxicants in an in vitro model of the BBB. These findings suggest that individuals who are exposed to environmental pollutants may suffer greater damage of their BBB if subsequently exposed to SARS‐CoV‐2 spike proteins or SARS‐CoV‐2.

Cornuside ameliorated experimental autoimmune encephalomyelitis by limiting the recruitment of CD4+ T lymphocytes in the spinal cord

We conducted this study to determine whether cornuside could improve the neurological deficit symptoms of experimental autoimmune encephalomyelitis (EAE) rats, as well as determine the potential involvement of CD4+ T lymphocytes, vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and tumor necrosis factor-α (TNF-α). Altogether, 32 Lewis rats were randomly divided into control, EAE, EAE/prednisolone, and EAE/cornuside, wherein their neurological function was assessed every day. CD4+ T lymphocyte recruitment into the spinal cord (SC) was evaluated using immunohistochemistry. The VCAM-1, ICAM-1 and TNF-α mRNA expressions in the SC were determined by real-time quantitative PCR, and the VCAM-1 and ICAM-1 proteins were determined by western blotting. Compared to the control group, the EAE group rats with neurological deficits had enhanced CD4+ T lymphocyte infiltration and higher expression levels of VCAM-1, ICAM-1, and TNF-α in the SC. Meanwhile, compared with the EAE group, the EAE/cornuside and EAE/prednisolone groups had lower neurological scores, less CD4+ T lymphocyte infiltrations, and lower expression levels of VCAM-1, ICAM-1, and TNF-α in the SC. Thus, cornuside ameliorated EAE, which could be owed to the inhibition of CD4+ T lymphocyte recruitment and VCAM-1, ICAM-1, and TNF-α expressions in the SC.

HO‑1 knockdown upregulates the expression of VCAM‑1 to induce neutrophil recruitment during renal ischemia‑reperfusion injury.

Heme oxygenase-1 (HO-1) has been reported to be upregulated following renal ischemia-reperfusion injury (IRI) and plays a key cytoprotective role; however, the underlying molecular mechanisms of its protective effects remain poorly understood. In the present study, in order to further elucidate the molecular mechanisms underlying the cytoprotective role of HO-1 in renal IRI, HO-1+/+ and HO-1+/− mice were subjected to renal ischemia and subsequent reperfusion followed by the analysis of blood urea nitrogen (BUN) and serum creatinine (SCr) levels, the severity of histological changes, HO-1 and vascular cell adhesion molecule-1 (VCAM-1) protein expression, the mRNA expression of inflammatory factors and the effects of VCAM-1 blockade. The results of the present study demonstrated that the upregulated expression levels of VCAM-1 in HO-1+/− mice during IRI increased the extent of renal tissue damage and activated the inflammatory response. These effects were subsequently reversed following infusion with an anti-VCAM-1 antibody. In addition, the upregulated expression of VCAM-1 in mouse glomerulus vascular endothelial cells isolated from HO-1+/− mice increased the adhesion and migration of neutrophils, effects which were also reversed upon incubation with an anti-VCAM-1 antibody. These results indicated that HO-1 knockdown may upregulate the expression of VCAM-1 during renal IRI, resulting in increased neutrophil recruitment and the activation of the inflammatory response, thereby exacerbating renal IRI. The present study thus highlights the regulatory mechanisms of HO-1 in renal IRI and provides a potential target for the clinical treatment of IRI following renal transplantation.

In Vitro Acute Ischemic Injury Altered Human Brain Endothelial MMP‐9 Activity and Associated Cell Surface Adhesion Protein Expression in Part by S1PR Type 1 Activation

Cerebrovascular endothelial cells are among the first cells to be impacted by acute ischemic stroke (AIS) which poses them as important therapeutic targets pivotal to the maintenance of blood brain barrier (BBB) integrity. Following AIS, matrix metalloproteinase 9 (MMP-9) proteolytic activity contributes to the loss in BBB integrity, increased hemorrhagic risk, and worse outcomes. We have previously shown that sphingosine-1-phosphate receptor (S1PR) ligands attenuated hypoxia plus glucose deprivation (HGD)-induced decreases in brain microvascular endothelial trans-endothelial resistance (TEER), suggesting a potential role for S1PR ligands on barrier function during AIS. Endothelial vascular cell adhesion molecule 1 (VCAM-1) is an inflammatory mediator and adhesion molecule that contributes to the loss of endothelial barrier integrity by serving as a scaffold for immune cell extravasation as well as a mediator for intracellular signals involved in MMP-9 activation. Cell surface adhesion receptor, CD44, likewise plays a key role in facilitating MMP-9 activity via co-localization at the endothelial surface. Thusly, we sought to investigate protein expression levels of VCAM-1, MMP-9, tight junctional proteins, (ZO-1 and claudin-5), as well as to characterize CD44 protein expression in cerebrovascular endothelial cells following HGD. Pediatric male human brain microvascular endothelial cells (HBMECs) were treated with ozanimod (selective S1PR type 1 ligand; 0.5 nM) at P7 and immediately exposed to normoxia (21% O2) or HGD (1% O2) for 3h, a timepoint determined from our previous TEER studies which revealed deterioration of the endothelial barrier as early as 3h. To verify S1PR type 1 dependency, selective blockade with antagonist W146 was applied to select experiments. We observed through standard immunoblotting that HGD induced a simultaneous decrease in ZO-1 and claudin-5 in parallel to increased expression of VCAM-1. Additionally, intracellular pro-MMP-9 (inactive enzymatic form) levels decreased following HGD and the ratio of active MMP-9 to pro-MMP-9 was increased. Ozanimod did not alter these responses. Using immunocytochemistry, anti-CD44 fluorescence was increased following HGD exposure, and ozanimod attenuated this response. S1PR type 1 blockade demonstrated receptor dependence. In conclusion, HGD induced endothelial activation characterized by increased adhesion molecule levels and involvement of MMP-9 activity potentially contributing to tight junction protein loss. Furthermore, S1PR type 1 ligands may in part attenuate HGD-induced BBB integrity loss by modulating the bioavailability of MMP-9 to disrupt endothelial barrier integrity. Mechanism(s) by which S1PR type 1 ligands regulate MMP-9 activity and its associated cell surface adhesion proteins (e.g. CD44 and VCAM-1) following ischemic-like injury warrants further investigation.

Abstract P778: Hypertension and Vascular Cell Adhesion Molecule 1 Relate To % Change in National Institutes of Health Stroke Scale Score in Ischemic Stroke After Mechanical Thrombectomy

Introduction: The University of Kentucky Blood and Clot Thrombectomy Registry and Collaboration (BACTRAC) protocol utilizes thrombectomy to isolate intracranial (i.e. distal to thrombus) arterial blood and systemic (i.e. carotid) arterial blood from thrombectomy procedures to study stroke. Here, we investigate the relationship among Vascular Cell Adhesion Molecule 1 (VCAM1), hypertension (HTN), and stroke recovery in patients undergoing mechanical thrombectomy for emergent large vessel occlusion (ELVO) stroke. Methods: Intracranial and systemic plasma samples from 25 subjects underwent cardiometabolic proteomic analysis at Olink Proteomics. Demographic data including HTN status and both admission NIHSS and discharge NIHSS were included. Linear regression analysis was run on both intracranial and systemic VCAM1 expression levels against % change in NIHSS ((Admittance NIHSS - Discharge NIHSS)/Admittance NIHSS)) and two-tailed t-tests were run assessing VCAM1 expression with HTN vs. no HTN. Results: Increased expression of intracranial VCAM1 significantly correlated with a smaller % change in NIHSS (p=0.001). Similarly, increased systemic VCAM1 expression was also found to have a significant relationship with smaller % change in NIHSS (p=0.005). Subjects with hypertension had significantly higher intracranial (p=0.03) and systemic (p=0.001) VCAM1 levels compared to those without HTN. Discussion: VCAM1 mediates leukocyte-endothelial cell adhesion and has been shown to play a role in stroke. This study takes a novel approach of sampling both intracranial and systemic arterial blood during an ELVO stroke. We found increased intracranial and systemic VCAM1 independently correlate with a smaller % change in NIHSS, an indicator of poorer initial recovery. Although preliminary, these results suggest an informative role of VCAM1 levels at the time of infarct. Additionally, those with HTN had higher levels of intracranial and systemic VCAM1. These data are in line with previous studies suggesting VCAM1 is a marker of endothelial damage due to HTN leading to negative clinical outcomes. To better understand our findings, we plan to perform subset analyses investigating VCAM1 levels in relation to dyslipidemia, infarct time and infarct volume.

Cilostazol on the expression of ICAM-1, VCAM-1 and inflammatory factors in plasma in patients with thromboangiitis obliterans.

The effects of cilostazol on the expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and inflammatory factors in plasma in patients with thromboangiitis obliterans (TAO) were studied. Plasma viscosity, fibrinogen, total cholesterol (TC) and triglyceride (TG) were detected for the healthy control, TAO and cilostazol groups, respectively. Results showed that compared with those in the control group, the plasma viscosity, fibrinogen, TC and TG levels in TAO group were significantly increased. By contrast, compared with those in TAO group, the plasma viscosity, fibrinogen, TC and TG levels in the cilostazol group were significantly decreased. ELISA results revealed that ICAM-1 and VCAM-1 expression levels in TAO group were obviously increased compared with those in control group. ICAM-1 and VCAM-1 expression levels in cilostazol group were obviously decreased compared with those in TAO group. According to RT-PCR, the mRNA expression levels of IL-1β, IL-6 and TNF-α in TAO group were significantly higher than those in control group, while the levels in cilostazol group were significantly decreased compared with those in TAO group. In addition, RT-PCR and western blotting proved that expression of both mRNA and protein of ICAM-1 and VCAM-1 in TAO group was significantly increased and obviously decreased after administration of cilostazol. The results of analysis of variance showed that the differences of ICAM-1 and VCAM-1 expression was statistically significant among the control, TAO and cilostazol groups (p<0.01). Cilostazol can significantly reduce the TAO-induced abnormal increase in ICAM-1, VCAM-1 and inflammatory factor expression in plasma in patients. It was proven that cilostazol has a good anti-TAO effect.

Soy-Leaf Extract Exerts Atheroprotective Effects via Modulation of Krüppel-Like Factor 2 and Adhesion Molecules

Soy-leaf extracts exert their cardioprotective effects by inducing endothelium-dependent vasodilation in the arteries, and they favorably modulate the serum lipid profile. In this study, we investigated the atheroprotective effects of an ethanol extract of soy leaf (ESL) in human umbilical vein endothelial cells (HUVECs) and high-cholesterol diet (HCD)-fed low-density lipoprotein receptor deficient (LDLR−/−) mice. ESL induced the expression of Krüppel-like factor 2 (KLF2), an endothelial transcription factor, and endothelial nitric oxide synthase (eNOS), and suppressed the expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) through moderate inflammatory signal activation, not only in tumor necrosis factor-α (TNF-α)-stimulated HUVECs but also in 7-ketocholesterol (7-KC)-stimulated HUVECs. ESL supplementation reduced aortic lesion formation in Western diet-fed LDLR−/− mice by 46% (p < 0.01) compared to the HCD group. ESL also markedly decreased the aortic expression levels of VCAM-1, ICAM-1, monocyte chemotactic protein-1 (MCP-1), TNF-α, IL-6, IL-1β, matrix metallopeptidase 9 (MMP-9), and fractalkine, while the expression of KLF2 was significantly increased. These results suggest that ESL supplementation has potential for preventing HCD-induced atherosclerosis effectively.

Abstract 1174: VCAM-1 enhances immune evasion in a murine model of metastatic cervical cancer

Abstract Surface over-expression of Vascular Cell Adhesion Molecule-1 (VCAM-1) has recently been identified as a mechanism used by metastatic breast cancer cells to provide pro-survival advantages. It has been previously shown that the metastatic and immune-resistant mouse cervical cancer cell line, A17 has a significantly increased expression of VCAM-1 compared with the primary parental tumor cell line, TC1-P0. Using this metastatic tumor model, we first modulated the surface expression levels of VCAM-1 on A17 in vitro by using an shRNA knockdown approach. We then inoculated these cells intra-venously (iv) and subcutaneously (sq) into C57BL/6 background mice and athymic nude mice to interrogate how VCAM-1 expression affects surrounding immune cell interactions within the tumor microenvironment. To further characterize VCAM-1 dependent immune responses, we used intravital 2-photon laser-scanning microscopy (2P-LPS) to dynamically visualize immune-tumor cells interaction. We observed that VCAM-1 down-regulation reduces the primary tumor growth kinetics in C57BL/6 mice but exhibited similar growth kinetics in nude mice, suggesting that VCAM-1 levels are not responsible for the intrinsic primary tumor growth differences in vivo. Moreover, VCAM-1 down-regulated cell lines showed decreased lung metastasis in both immune-competent mice and athymic nude mice. Furthermore, the metastatic lung parenchyma in mice injected with the non-silencing shVCAM-1 control exhibited immune cell infiltrates with a predominance of M2 polarized macrophages and an altered cytokine production. This may be responsible for decreased susceptibility to immune surveillance. Together, our results demonstrate that tumor-associated VCAM-1 expression helps aggressive tumors circumvent effective immune surveillance, and may represent a target for the development of immune-mediated therapy in metastatic cervical cancer. Citation Format: Francesca Scrimieri, Iuliana Dit Bobanga, Saada Eid, David J. Corn, Deborah Barkauskas, Jay Jay Myers, Alex Y. Huang. VCAM-1 enhances immune evasion in a murine model of metastatic cervical cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1174. doi:10.1158/1538-7445.AM2014-1174

99mTc-cAbVCAM1-5 imaging is a sensitive and reproducible tool for the detection of inflamed atherosclerotic lesions in mice.

(99m)Tc-cAbVCAM1-5, a single-domain antibody fragment directed against mouse or human vascular cell adhesion molecule 1 (VCAM-1), recently has been proposed as a new imaging agent for the detection of inflamed atherosclerotic lesions. Indeed, in a mouse model of atherosclerosis, (99m)Tc-cAbVCAM1-5 specifically bound to VCAM-1-positive lesions, thereby allowing their identification on SPECT images. The purpose of the present study was to investigate (99m)Tc-cAbVCAM1-5 imaging sensitivity using a reference statin therapy. Thirty apolipoprotein E-deficient mice were fed a western-type diet. First, the relationship between the level of VCAM-1 expression and (99m)Tc-cAbVCAM1-5 uptake was evaluated in 18 mice using immunohistochemistry and autoradiography. Second, longitudinal SPECT/CT imaging was performed on control (n = 9) or atorvastatin-treated mice (0.01% w/w, n = 9). (99m)Tc-cAbVCAM1-5 uptake in atherosclerotic lesions correlated with the level of VCAM-1 expression (P < 0.05). Atorvastatin exerted significant antiatherogenic effects, and (99m)Tc-cAbVCAM1-5 lesion uptake was significantly reduced in 35-wk-old atorvastatin-treated mice, as indicated by ex vivo γ-well counting and autoradiography (P < 0.05). SPECT imaging quantification based on contrast-enhanced CT was reproducible (interexperimenter intraclass correlation coefficient, 0.97; intraexperimenter intraclass correlation coefficient, 0.90), and yielded results that were highly correlated with tracer biodistribution (r = 0.83; P < 0.0001). Therefore, reduced (99m)Tc-cAbVCAM1-5 uptake in atorvastatin-treated mice was successfully monitored noninvasively by SPECT/CT imaging (0.87 ± 0.06 vs. 1.11 ± 0.09 percentage injected dose per cubic centimeter in control group, P < 0.05). (99m)Tc-cAbVCAM1-5 imaging allowed the specific, sensitive, and reproducible quantification of VCAM-1 expression in mouse atherosclerotic lesions. (99m)Tc-cAbVCAM1-5 therefore exhibits suitable characteristics for the evaluation of novel antiatherogenic agents.

MONITORING THE PROGRESSION OF ATHEROSCLEROSIS IN RABBITS MODEL WITH TARGETED ULTRASOUND DETECTION OF VASCULAR CELL ADHESION MOLECULE-1

ObjectivesThe aim of our study was to investigate the use of targeted contrast-enhanced high-frequency ultrasonography for molecular imaging of vascular cell adhesion molecule-1 (VCAM-1) expression on carotid artery in a...

Anti-atherosclerotic function of Astragali Radix extract: downregulation of adhesion molecules in vitro and in vivo

BackgroundAtherosclerosis is considered to be a chronic inflammatory disease. Astragali Radix extract (ARE) is one of the major active ingredients extracted from the root of Astragalus membranaceus Bge. Although ARE has an anti-inflammatory function, its anti-atherosclerotic effects and mechanisms have not yet been elucidated.MethodsMurine endothelial SVEC4-10 cells were pretreated with different doses of ARE at different times prior to induction with tumor necrosis factor (TNF)-α. Cell adhesion assays were performed using THP-1 cells and assessed by enzyme-linked immunosorbent assay, western blotting and immunofluorescence analyses to detect the expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), phosphorylated inhibitor of κB (p-iκB) and nuclear factor (NF)-κB. We also examined the effect of ARE on atherosclerosis in the aortic endothelium of apolipoprotein E-deficient (apoE−/−) mice.ResultsTNF-α strongly increased the expression of VCAM-1 and ICAM-1 accompanied by increased expression of p-iκB and NF-κB proteins. However, the expression levels of VCAM-1 and ICAM-1 were reduced by ARE in dose- and time-dependent manners, with the strongest effect at a dose of 120 μg/ml incubated for 4 h. This was accompanied by significantly decreased expression of p-iκB and inhibited activation of NF-κB. Immunofluorescence analysis also revealed that oral administration of ARE resulted in downregulation of adhesion molecules and decreased expression of macrophages in the aortic endothelium of apoE−/− mice. ARE could suppress the inflammatory reaction and inhibit the progression of atherosclerotic lesions in apoE−/− mice.ConclusionThis study demonstrated that ARE might be an effective anti-inflammatory agent for the treatment of atherosclerosis, possibly acting via the decreased expression of adhesion molecules.

Anti-atherosclerotic Activity of Platycodin D Derived from Roots of &lt;i&gt;Platycodon grandiflorum&lt;/i&gt; in Human Endothelial Cells

This study examined the effects of platycodin D (PD), a triterpene saponin from the the root of Platycodon grandiflorum A.DC on human umbilical vein endothelial cells (HUVECs) in vitro, which were pre-treated with PD (0.01, 0.15, 0.25 mg/mL), respectively, and treated with 50 mg/L oxidized low-density lipoprotein (oxLDL). The levels of nitric oxide (NO) and malonaldehyde (MAD) in the culture medium, vascular cell adhesion molecule-1 (VCAM-1) and intercellular cell adhesion molecule-1 (ICAM-1) mRNA expression in endothelium cells and the adhesion of monocytes to endothelial cells were measured. The results showed that PD increased NO concentration and decreased MDA level induced by oxLDL in the medium of endothelial cells. Moreover, PD significantly inhibited the oxLDL-induced increase in monocyte adhesion to endothelial cells as well as decreasing mRNA expression levels of VCAM-1 and ICAM-1 on these cells. Based on these results, it is suggested that PD is a promising anti-atherosclerotic activity, which is at least in part the result of its increasing NO concentration, reducing the oxLDL-induced cell adhesion molecule expression in endothelial cells and the endothelial adhesion to monocytes.

Effects of Angiotensin II and Shear Stress Interaction on Vascular Inflammation

❙ABSTRACT❙ Background: Angiotensin II (AngII) and abnormal oscillatory shear stress are highly associated with vascular inflammation including atherosclerosis. However, it is poorly understood how interactions between AngII and shear stress in human aortic endothelial cells (HAEC) are involved in mechanisms by which cellular adhesion molecules are expressed. The purpose of this study was to improve that understanding. Methods: AngII (10 -7 M for 6 hr) and two-types of shear stress treatments were used: laminar shear stress (LS: unidirectional, 12 dynes/cm 2 ) and oscillatory shear stress (OS: bi-directional, 5 dynes/cm 2 , 1 Hz) in HAEC. Immunoblotting was used to detect expression of cellular adhesion molecules markers such as vascular cell adhesion molecule 1 (VCAM1) and intercellular adhesion molecule 1 (ICAM1). Results: AngII significantly increased VCAM1 and ICAM1 expression in HAEC that had been reduced due to pretreatment with telmisartan. AngII-LS co-stimulation and AngII-OS co-stimulation significantly increased VCAM1 and ICAM1 expression in HAEC. The expression levels of VCAM1 and ICAM1 were also, significantly reduced when pretreated with telmisartan. However, VCAM1 and ICAM1 expression were significantly reduced under LS and OS stimulation. Conclusions: Telmisartan may modulate the expressions of VCAM1 and ICAM1 via different types of shear stress in HAEC that are activated by AngII. (J Korean Soc Hypertens 2011;17(1):17-27)

Polychlorinated biphenyl-induced VCAM-1 expression is attenuated in aortic endothelial cells isolated from caveolin-1 deficient mice

Exposure to environmental contaminants, such as polychlorinated biphenyls (PCBs), is a risk factor for the development of cardiovascular diseases such as atherosclerosis. Vascular cell adhesion molecule-1 (VCAM-1) is a critical mediator for adhesion and uptake of monocytes across the endothelium in the early stages of atherosclerosis development. The upregulation of VCAM-1 by PCBs may be dependent on functional membrane domains called caveolae. Caveolae are particularly abundant in endothelial cell membranes and involved in trafficking and signal transduction. The objective of this study was to investigate the role of caveolae in PCB-induced endothelial cell dysfunction. Primary mouse aortic endothelial cells (MAECs) isolated from caveolin-1-deficient mice and background C57BL/6 mice were treated with coplanar PCBs, such as PCB77 and PCB126. In addition, siRNA gene silencing technique was used to knockdown caveolin-1 in porcine vascular endothelial cells. In MAECs with functional caveolae, VCAM-1 protein levels were increased after exposure to both coplanar PCBs, whereas expression levels of VCAM-1 were not significantly altered in cells deficient of caveolin-1. Furthermore, PCB-induced monocyte adhesion was attenuated in caveolin-1-deficient MAECs. Similarly, siRNA silencing of caveolin-1 in porcine endothelial cells confirmed the caveolin-1-dependent VCAM-1 expression. Treatment of cells with PCB77 and PCB126 resulted in phosphorylation of extracellular signal-regulated kinase-1/2 (ERK1/2), and pharmacological inhibition of ERK1/2 diminished the observed PCB-induced increase in monocyte adhesion. These findings suggest that coplanar PCBs induce adhesion molecule expression, such as VCAM-1, in endothelial cells, and that this response is regulated by caveolin-1 and functional caveolae. Our data demonstrate a critical role of functional caveolae in the activation and dysfunction of endothelial cells by coplanar PCBs.

Prevention of allograft vasculopathy by transfection of calcitonin gene-related peptide

To explore suppression of allograft vasculopathy (AV) by transfer of the calcitonin gene-related peptide (CGRP). The descending thoracic aortas from 39 Lewis rats were grafted to the abdominal aortas of 39 F344 rats, and then the recipient rats were randomized into 3 groups: Group A transfected with Ad5-CGRP-EGFP, a gene construct containing sequences from the adenoviral oncoprotein, CGRP, and enhanced green fluorescent protein, Group B, transfected with Ad5-EGFP containing sequences from the adenoviral oncoprotein and enhanced green fluorescent protein, and Group C without transfection. Four and 8 weeks later the abdominal aortas of 6 recipient rats from each were collected respectively. The degree of vascular obstruction was observed by microscopy. Frozen tissue sections were made and the inverted phase contrast fluorescence microscopy was used to observe the green fluorescence showing the expression of CGRP and enhanced green fluorescent protein (EGFP). RT-PCR was used to detect the expression of CGRP. Immunohistochemistry was used to examine the expression of inducible nitric oxide synthase (iNOS) and vascular cell adhesion molecule-1 (VCAM-1). The apoptosis index (AI) was detected by TUNEL method. CGRP expression was positive in both Group A and B 4 weeks later and negative in both groups 8 weeks later. Group A is approximately normal in pathological morphology 4 weeks later. The vascular luminal occlusion score of Group A was lower than Groups B and C 4 weeks later, and significantly higher than that I the same group (P < 0.05), however, still lower than those of Groups B and C 8 weeks later. The AI 4 weeks later of Group A was significantly lower than those of Groups B and group C, however, there was no significance in AI among the 3 groups. Four weeks later the VCAM-1 expression levels in the tunicae intima, media, and adventitia of Group A were all significantly lower than those of Group B C 4 weeks later, however, the iNOS expression level of Group A was only significantly lower than those of Groups B and C in the tunica intima 4 weeks later. The expression of CGRP effectively suppresses the development of AV 4 weeks after the operation.

Renal allograft rejection: expression and function of VCAM-1 on tubular epithelial cells.

The interaction between vascular cell adhesion molecule-1 (VCAM-1) and very late antigen-4 (VLA-4) is known to play an important role in stabilizing the adhesion of lymphocytes to endothelial cells. Such cellular adhesion is crucial to many immunological processes including lymphocyte-mediated cell lysis. In this study the expression of VCAM-1 on renal tubular epithelial cells is demonstrated on biopsy sections recovered during acute renal allograft rejection. Experiments performed using epithelial cells cultured from renal tubules show that VCAM-1 is up-regulated by addition of the inflammatory cytokines tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). Combination of TNF-alpha and IFN-gamma synergized to induce high levels of VCAM-1 expression. Further experiments demonstrated that the cytokines produced by activated lymphocytes in mixed leucocyte culture also up-regulate expression of VCAM-1. Assays of the adhesion of lymphoid cells to cultured renal epithelial cells showed that cytokine pretreatment of the renal cells enhanced the binding of lymphoid cells. The proportion of bound lymphoid cells was significantly reduced by addition of an antibody capable of blocking the interaction of VCAM-1 with VLA-4. This result indicated that the VCAM-1 induced on renal epithelial cells by inflammatory cytokines is functionally capable of binding VLA-4, thereby enhancing the adhesion of potentially graft-damaging lymphoid cells.

Vascular Cell Adhesion Molecule 1 Predicts Cancer-Free Survival in Clear Cell Renal Carcinoma Patients

Vascular cell adhesion molecule 1 (VCAM1) is a cell surface glycoprotein implicated in various pathophysiologic conditions. We measured VCAM1 expression levels in tumor tissues and evaluated its significance and prognostic use in renal cell carcinoma (RCC). We used real-time quantitative PCR to examine the VCAM1 expression levels of a total of 485 sporadic renal tumors, including 429 clear cell, 21 papillary, 17 chromophobe, 11 oncocytomas, and 7 collecting duct carcinomas. We retrospectively examined the relationship of this expression to various clinicopathologic variables and the von Hippel-Lindau alteration status. We evaluated its significance with respect to patient survival rates using the Cox regression model combined with the split-sample method. Compared with normal kidney samples (n = 43), VCAM1 was significantly up-regulated in clear cell RCC and papillary RCC, whereas it was down-regulated in chromophobe RCC and oncocytoma. In clear cell RCC, VCAM1 expression levels were apparently high in patients asymptomatic at presentation and in patients with small tumor size, low-stage, low-grade, microvascular invasion-negative, and von Hippel-Lindau alteration-positive tumors. Univariate analyses showed that VCAM1 high expression is strongly associated with better outcomes in clear cell and papillary RCCs. Further, Cox multivariate analysis models combined with the split-sample method revealed that this association is significant only in cancer-free survival for patients with clear cell RCC after curative surgical resection. VCAM1 expression levels were found to be histologically subtype specific in renal tumors. Determination of the VCAM1 expression level as a biomarker can provide useful prognostic information for patients with clear cell RCC.