Intercellular Adhesion Molecule-1 Levels Research Articles

A Allele of ICAM-1 Rs5498 and VCAM-1 Rs3181092 is Correlated with Increased Risk for Periodontal Disease.

ObjectivePeriodontal disease (PD) is viewed today as multifactorial problems initiated and sustained by bacteria but significantly modified by the body’s response to bacterial plaque. Recent studies have suggested that gene polymorphisms could be involved in the pathophysiology of periodontitis. This study aimed to investigate a possible correlation of the polymorphisms of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) with PD.MethodsThe genotypes of ICAM-1 and VCAM-1 were initially determined in PD patients using denaturing high performance liquid chromatography (DHPLC). ELISA was then conducted to measure ICAM-1 and VCAM-1 protein levels. Next, the association of ICAM-1/VCAM-1 genotype distribution and expression with clinical indicators and severity of PD was analyzed.ResultsPD patients contained increased levels of hemoglobin A1c (HbA1c), total cholesterol (TC), triglyceride (TG), and low-density lipoprotein (LDL), increased ICAM-1 and VCAM-1 protein levels, and decreased high-density lipoprotein (HDL) level. The GG genotype and G allele at ICAM-1 rs5498, as well as the AG and GG genotypes and G allele at VCAM-1 rs3181092 may reduce PD risk.ConclusionTo sum up, the overexpressed ICAM-1 and VCA M-1 as well as A allele of ICAM-1 rs5498 and VCAM-1 rs3181092 is associated with the onset of PD.

Rice-derived peptide AAGALPS inhibits TNF-α-induced inflammation and oxidative stress in vascular endothelial cells.

Injury of vascular endothelial cell is one of the main factors triggering atherosclerosis. Peptide AAGALPS was derived from digestion and absorption product of rice α‐globulin, which was proved to prevent atherosclerosis in previous study. This study aims to investigate the potential effects of AAGALPS on improving tumor necrosis factor‐α (TNF‐α)‐stimulated human umbilical vein endothelial cells’ (HUVECs) injury. As a result, the viability of HUVECs stimulated by tumor necrosis factor‐α was significantly increased by AAGALPS in a dose‐dependent manner until 25 μg/ml. The peptide obviously reduced the levels of intercellular adhesion molecule‐1, vascular cell adhesion molecule‐1, nitric oxide, inducible nitric oxide synthase, reactive oxygen species, and malondialdehyde and increased the concentrations of glutathione peroxidase. Furthermore, AAGALPS inhibited the nuclear factor κB (NF‐κB) activation and nuclear translocation through regulating inhibitor of nuclear factor κB kinase α and inhibitor of NF‐κB. These results indicated that AAGALPS protected vascular endothelial cells through mediating inflammation and oxidative stress.

Proinflammatory Mediators, IL (Interleukin)-1β, TNF (Tumor Necrosis Factor) α, and Thrombin Directly Induce Capillary Tube Regression.

In this work, we examine the molecular basis for capillary tube regression and identify key proregressive factors, signaling pathways, and pharmacological antagonists of this process. Approach and Results: We demonstrate that the proinflammatory mediators, IL (interleukin)-1β, TNF (tumor necrosis factor) α, and thrombin, singly and in combination, are potent regulators of capillary tube regression in vitro. These proregressive factors, when added to endothelial cell-pericyte cocultures, led to selective loss of endothelial cell-lined tube networks, with retention and proliferation of pericytes despite the marked destruction of adjacent capillary tubes. Moreover, treatment of macrophages with the TLR (toll-like receptor) agonists Pam3CSK4 and lipopolysaccharide generates conditioned media with marked proregressive activity, that is completely blocked by a combination of neutralizing antibodies directed to IL-1β and TNFα but not to other factors. The same combination of blocking antibodies, as well as the anti-inflammatory cytokine IL-10, interfere with macrophage-dependent hyaloid vasculature regression in mice suggesting that proinflammatory cytokine signaling regulates capillary regression in vivo. In addition, we identified a capillary regression signaling signature in endothelial cells downstream of these proregressive agents that is characterized by increased levels of ICAM-1 (intercellular adhesion molecule-1), phospho-p38, and phospho-MLC2 (myosin light chain-2) and decreased levels of phospho-Pak2, acetylated tubulin, phospho-cofilin, and pro-caspase3. Finally, we identified combinations of pharmacological agents (ie, FIST and FISTSB) that markedly rescue the proregressive activities of IL-1β, TNFα, and thrombin, individually and in combination. Overall, these new studies demonstrate that the major proinflammatory mediators, IL-1β, TNFα, and thrombin, are key regulators of capillary tube regression-a critical pathological process regulating human disease.

Roseltide rT7 is a disulfide-rich, anionic, and cell-penetrating peptide that inhibits proteasomal degradation

Disulfide-rich plant peptides with molecular masses of 2-6 kDa represent an expanding class of peptidyl-type natural products with diverse functions. They are structurally compact, hyperstable, and underexplored as cell-penetrating agents that inhibit intracellular functions. Here, we report the discovery of an anionic, 34-residue peptide, the disulfide-rich roseltide rT7 from Hibiscus sabdariffa (of the Malvaceae family) that penetrates cells and inhibits their proteasomal activities. Combined proteomics and NMR spectroscopy revealed that roseltide rT7 is a cystine-knotted, six-cysteine hevein-like cysteine-rich peptide. A pair-wise comparison indicated that roseltide rT7 is >100-fold more stable against protease degradation than its S-alkylated analog. Confocal microscopy studies and cell-based assays disclosed that after roseltide rT7 penetrates cells, it causes accumulation of ubiquitinated proteins, inhibits human 20S proteasomes, reduces tumor necrosis factor-induced IκBα degradation, and decreases expression levels of intercellular adhesion molecule-1. Structure-activity studies revealed that roseltide rT7 uses a canonical substrate-binding mechanism for proteasomal inhibition enabled by an IIML motif embedded in its proline-rich and exceptionally long intercysteine loop 4. Taken together, our results provide mechanistic insights into a novel disulfide-rich, anionic, and cell-penetrating peptide, representing a potential lead for further development as a proteasomal inhibitor in anti-cancer or anti-inflammatory therapies.

Skeletal stem cell-mediated suppression on inflammatory osteoclastogenesis occurs via concerted action of cell adhesion molecules and osteoprotegerin

In the current study, we investigated how skeletal stem cells (SSCs) modulate inflammatory osteoclast (OC) formation and bone resorption. Notably, we found that intercellular adhesion molecule‐1 (ICAM‐1), vascular cell adhesion molecule‐1 (VCAM‐1), and osteoprotegerin (OPG) play a synergistic role in SSC‐mediated suppression of inflammatory osteoclastogenesis. The effect of SSCs on inflammatory osteoclastogenesis was investigated using a lipopolysaccharide‐induced mouse osteolysis model in vivo and human osteoarthritis synovial fluid (OASF) in vitro. OC formation was determined by tartrate‐resistant acid phosphatase staining. Bone resorption was evaluated by microcomputerized tomography, serum C‐terminal telopeptide assay, and pit formation assay. The expression of ICAM‐1, VCAM‐1, and OPG in SSCs and their contribution to the suppression of osteoclastogenesis were determined by flow cytometry or enzyme linked immunosorbent assay. Gene modification, neutralization antibodies, and tumor necrosis factor‐α knockout mice were used to further explore the mechanism. The results demonstrated that SSCs remarkably inhibited inflammatory osteoclastogenesis in vivo and in vitro. Mechanistically, inflammatory OASF stimulated ICAM‐1 and VCAM‐1 expression as well as OPG secretion by SSCs. In addition, ICAM‐1 and VCAM‐1 recruited CD11b+ OC progenitors to proximity with SSCs, which strengthened the inhibitory effects of SSC‐derived OPG on osteoclastogenesis. Furthermore, it was revealed that tumor necrosis factor α is closely involved in the suppressive effects. In summary, SSCs express a higher level of ICAM‐1 and VCAM‐1 and produce more OPG in inflammatory microenvironments, which are sufficient to inhibit osteoclastogenesis in a “capture and educate” manner. These results may represent a synergistic mechanism to prevent bone erosion during joint inflammation by SSCs.

Perilla frutescens Britton var. frutescens leaves attenuate dextran sulfate sodium-induced acute colitis in mice and lipopolysaccharide-stimulated angiogenic processes in human umbilical vein endothelial cells

The aim of the current study was to investigate whether the leaves of Perilla frutescens Britton var. frutescens (PL), a frequently consumed vegetable in Korea, attenuate dextran sulfate sodium (DSS)-induced acute colitis in mice and lipopolysaccharide (LPS)-stimulated angiogenic processes in human umbilical vein endothelial cells (HUVEC). In DSS-treated mice, dietary supplementation with PL mitigated DAI and colon shortening. The dietary PL also reduced colonic levels of inflammatory and angiogenic mediators, such as interleukin-1β, interleukin-6, monocyte chemoattractant protein-1, macrophage inflammatory protein-2, leukotriene B4, inducible nitric oxide synthase, cyclooxygenase-2, basic fibroblast growth factor, and intercellular adhesion molecule-1 (ICAM-1). Treatment of HUVEC with ethanol extract of PL attenuated LPS-stimulated increases in ICAM-1 levels, monocyte adhesion, invasion, and tube formation. This study suggests that dietary PL effectively inhibited DSS-induced acute colitis in mice, and its anti-angiogenic activities may partially contribute to the inhibition.

Evaluation of intercellular adhesion molecule-1 (ICAM-1), Tumor necrosis factor α (TNF-α), Interleukins (IL-6, IL-8) and C-reactive protein (CRP) levels in neonatal calves with presumed septicemia

In this study; intercellular molecule-1 (ICAM-1), Tumor necrosis factor α (TNF-α), Interleukin-6 (IL-6), Interleukin-8 (IL-8) and C-reactive protein (CRP) levels were evaluated in neonatal calves with presumed septicemia. The material of this study consisted of a total of 30 calves. Study group consisted of 20 septicemic calves and 10 clinically healthy calves and with same breed and weight and their ages were between 1 and 10 days. The statistical analysis indicated that leukocyte (WBC), red blood cell (RBC), hematocrit (Hct) hemoglobin (Hb) and mean corpuscular haemoglobin concentration (MCHC) values in neonatal calves with presumed septicemia were higher than those of the control group (P<0.05), whereas PLT values of neonatal calves with septicemia were lower than those of the control group (P<0.05). The levels of ICAM-1, TNF-α, IL-6, IL-8 and CRP values in septicemic calves were higher than those of the control group (P<0.05). As a result, it was concluded that analyzes of TNF-α, IL-6, IL-8 and particularly ICAM-1 concentrations may be considered in evaluating the severity and prognosis of patients with septicemia, as well as monitoring the efficacy of the treatment of the disease.

The Role of Intercellular Adhesion Molecule-1 in the Pathogenesis of Psychiatric Disorders.

Intercellular adhesion molecule-1 (ICAM-1) is a transmembrane glycoprotein that is overexpressed in many pathological states. Although, like many other immune molecules, ICAM-1 plays only a limited role in the abundant concert of the immune response, it may be more important than we realize. In the central nervous system (CNS), ICAM-1 is expressed in microglial cells and astrocytes and in endothelial cells in the white and gray matter of the human forebrain. It is of particular interest in psychiatric disorders for two reasons: It has a key function for the blood–brain barrier, which plays an important role in the biology of psychiatric disorders, and it is a marker for inflammation. Although the blood level of soluble ICAM-1 (sICAM-1) might be lower in acute unmedicated schizophrenia, it has been reported to be increased in many other psychiatric conditions, such as major depression, bipolar disorder, and dementia. In bipolar disorder, high sICAM levels were found during both the depressed and the manic states and also during the euthymic phase (the free interval), possibly indicating that sICAM is a trait marker. High sICAM-1 blood levels have also been found in depression comorbid to a somatic disease state. Interestingly, sICAM-1 levels also increase during aging. Some studies investigated sICAM-1 levels in the cerebrospinal fluid of psychiatric disorders and ICAM-1 expression in postmortem CNS tissue of psychiatric patients and found that the overall duration and duration of the chronic phase of the psychiatric disorder seem to play a role in both. Moreover, confounders, such as antipsychotic and antidepressive medication, have to be considered. sICAM-1 levels seem to be associated with hypopermeability or hyperpermeability of the blood-brain barrier and thus to influence the communication between the CNS immune system, represented by glia cells, and the peripheral immune system. The balance between the influx and efflux of immune molecules into and out of the CNS may be one of the pinpoints in psychiatric disorders, in particular in the chronic phase, e.g., in schizophrenia. This aspect, however, needs further intense research, in particular to enable researchers to develop therapeutic principles based on an immune/inflammatory approach.

Pharmacological blockage of ICAM-1 improves angiotensin II-induced cardiac remodeling by inhibiting adhesion of LFA-1+ monocytes.

Intercellular adhesion molecule-1 (ICAM-1) is a member of an immunoglobulin-like superfamily of adhesion molecules that mediate leukocyte adhesion to vascular endothelium and are involved in several cardiovascular diseases, including ischemia-reperfusion injury, myocardial infarction, and atherosclerosis. However, the role of ICAM-1 in angiotensin II (ANG II)-induced cardiac remodeling in mice remains unclear. Wild-type mice were administered an IgG control or ICAM-1 neutralizing antibody (1 and 2 mg/mouse, respectively) and ANG II (1,000 ng·kg-1·min-1) for up to 14 days. Cardiac contractile function and structure were detected by echocardiography. Hypertrophy, fibrosis, and inflammation were assessed by histological examination. The infiltration of lymphocyte function-associated antigen-1 (LFA-1+) monocytes/macrophages was assessed by immunostaining. The mRNA expression of genes was evaluated by quantitative RT-PCR analysis. Protein levels were tested by immunoblotting. We found that ICAM-1 expression in ANG II-infused hearts and ICAM-1 levels in serum from human patients with heart failure were significantly increased. Moreover, ANG II infusion markedly enhanced ANG II-induced hypertension, caused cardiac contractile dysfunction, and promoted cardiac hypertrophy, fibrosis, and LFA-1+ macrophage infiltration. Conversely, blockage of ICAM-1 with a neutralizing antibody dose-dependently attenuated these effects. Moreover, our in vitro data further demonstrated that blocking ICAM-1 inhibited ANG II-induced LFA-1+ macrophage adhesion to endothelial cells and migration. In conclusion, these results provide novel evidence that blocking ICAM-1 exerts a protective effect in ANG II-induced cardiac remodeling at least in part through the modulation of adhesion and infiltration of LFA-1+ macrophages in the heart. Inhibition of ICAM-1 may represent a new therapeutic approach for hypertrophic heart diseases.NEW & NOTEWORTHY Leukocyte adhesion to vascular endothelium is a critical step in cardiovascular diseases. ICAM-1 is a member of immunoglobulin-like superfamily of adhesion molecules that binds LFA-1 to mediate leukocytes adhesion and migration. However, the significance of ICAM-1 in ANG II-induced cardiac remodeling remains unclear. This study reveals that blocking of ICAM-1 prevents ANG II-induced cardiac remodeling via modulating adhesion and migration of LFA-1+ monocytes, may serve as a novel therapeutic target for hypertensive cardiac diseases.

Greater IL-6, D-dimer, and ICAM-1 Levels Are Associated With Lower Small HDL Particle Concentration in the Multicenter AIDS Cohort Study.

Low HDL cholesterol (HDL-C) is common in people living with HIV infection, which is associated with inflammation, and correlates with greater cardiovascular disease (CVD) risk. Particles of HDL are HDL subfractions, and in some general population studies, higher small HDL particle number (HDL-P) has been associated with lower CVD risk. The objective of this study was to determine whether HIV serostatus and systemic inflammation were associated with small HDL-P in the Multicenter AIDS Cohort Study (MACS). The MACS is composed of HIV-infected and HIV-uninfected men. Separate linear regression analyses were conducted to evaluate the associations between outcomes (small HDL-P, large HDL-P, total HDL-P, and HDL size) and variables of interest (interleukin-6 [IL-6], D-dimer, and intercellular adhesion molecule-1 [ICAM-1] levels), with adjustment for other CVD risk factors. The study population included 553 HIV-infected (88.1% on current ART) and 319 HIV-uninfected men. The mean age was 52.7 years for HIV-infected men and 55.3 years for HIV-uninfected men. In separate models of the study population, higher log IL-6 was associated with lower total and small HDL-P (P < .01 for both), independent of HIV serostatus and CVD risk factors. Similar results were seen with ICAM-1. Positive HIV serostatus was associated with lower small and total HDL-P, adjusted for inflammatory markers. Greater systemic inflammation and HIV infection both were associated with lower atheroprotective small HDL-P. This may be a potential mechanism contributing to increased cardiovascular risk among HIV-infected people.

Plasma BDNF Levels Are Associated with Stroke in Children with SCD

Background: Cerebrovascular disease, particularly overt stroke, is one of the most critical clinical complications of sickle cell disease (SCD). Individual stroke risk for patients with SCD is assessed by measuring trans-cranial Doppler ultrasound (TCD) velocity. While TCD screening and resulting use of chronic blood transfusion therapy in patients with abnormal TCDs has reduced stroke incidence in patients with SCD from 11% to 1%, the test has significant limitations. It can only be performed on children old enough to remain still for the assessment, or young enough to have open bony windows; typically 2-16 years of age. Additionally the test has poor positive predictive value, causing patients to be placed on chronic transfusion therapy who would not have gone on to have a stroke. Hydroxyurea (HU) is replacing chronic transfusion therapy in many institutions for primary stroke prevention, but many sites initiate a "cooling off" period of transfusions prior to initiating HU, and transfusion therapy may be indicated for abnormal TCD velocities in patients already on HU. The pathophysiology of stroke in SCD patients is not completely understood, but vascular remodeling, abnormal cerebral blood flow, and vaso-occlusion all play a role. Plasma levels of Brain derived neurotrophic factor (BDNF), myeloperoxidase (MPO), vascular cell adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1 (ICAM-1) have been shown to be associated with high trans-cranial Doppler ultrasound (TCD) velocity in children with SCD; but there is limited data on their potential association with stroke in this population. BDNF is a neurotropin that is responsive to hypoxia and promotes neurogenesis as well as playing a key role in the regulation of the cell survival pathway; myeloperoxidase (MPO) promotes vascular injury by oxygen radical generation, leukocyte/neutrophil recruitment, and oxidative stress; vascular cell adhesion molecule 1 (VCAM-1) is and endothelial adhesion molecule involved in the pathophysiology of vaso-occlusion in SCD through promoting RBC adhesion; and intercellular adhesion molecule 1 (ICAM-1) is expressed on endothelial and immune system cells and is thought to be a ligand for leukocyte adhesion. We hypothesized that BDNF, MPO, VCAM-1 and ICAM-1 may be involved in stroke in SCD, and may be potential biomarkers to be used to assess an individual with SCD's stroke risk in addition to TCD. Methods: We collected plasma samples from the peripheral blood of fifteen SCD patients (HbSS) at the time of either silent cerebral infarct (SCI) or stroke. Stroke (n=8) or SI was confirmed by MRI using diffusion imaging and T2 FLAIR. The cohort included 6 females and 9 males between the ages of 3 and 20 years. Plasma from fifteen age and gender matched control patients with HbSS but had normal MRIs and TCD velocities less than 170 m/s. We measured plasma levels of BDNF, MPO, VCAM-1 and ICAM-1 using antibody immobilized fluorescent beads (Millipore, Billerica, MA) and Luminex xMAP technology (Bio-Rad, Hercules, CA). Student's t-test was used to analyze the difference in plasma levels between the stroke and control groups. Results and Conclusions: BDNF levels were significantly higher in the stroke group than in the control group; 2978.2 ± 960.3 pg/ml compared to 2200.8 ± 758.4 pg/ml, p=0.005. Difference in MPO levels between the two groups approached significance; 36617.2 ± 14828.9 pg/ml compared to 29521.9 ± 7889.6 pg/ml, p=0.07. Difference in VCAM-1 levels also approached significance; 143997.0 ± 9963.8 pg/ml compared to 138126.9 ± 11902.0 pg/ml, p=0.08, but there was no significant difference in ICAM-1 levels. Only BDNF plasma levels positively correlated with the area and volume of the infarct, p=0.009. These results further support the assertion that BDNF is involved in the pathophysiology of cerebrovascular disease in SCD. It is possible that MPO and VCAM-1 may also play a role, and that a significant difference was not found due to sample size limitations. Plasma BDNF levels, and possibly MPO and VCAM-1 plasma levels, have potential as biomarkers for stroke risk to improve the positive predictive value of TCD velocities in patients with SCD, or to assess risk in patients unable to receive TCDs. Disclosures No relevant conflicts of interest to declare.

The Relationship Between Global Sleep Score And Inflammatory Markers In Obese Adults From The United States.

BackgroundPoor sleep is a risk factor for cardiovascular diseases (CVDs). The underlying pathogenesis is not clear. Levels of inflammatory markers, such as C-reactive protein (CRP), interleukin-6 (IL-6) and tumor necrosis factor-ɑ (TNF-ɑ), have been found to be elevated in patients with CVDs.AimThe study aimed to investigate the associations between sleep quality and serum inflammatory markers in a cohort of obese adults.MethodsThis was a second analysis of the data from the Midlife in the United States (MIDUS) study, a longitudinal study of a national (US) sample of adults. A total of 1255 participants completed comprehensive biological assessments. The associations between global sleep score and serum levels of inflammatory markers were analyzed.ResultsUnivariate analysis showed that a higher global sleep score was correlated with lower age (r = −0.079, P= 0.009), higher BMI (r = 0.100, P= 0.001) and heavier perceived stress (r = 0.335, P<0.001). Multivariate linear regression analysis showed that the global sleep score was positively related to levels of IL-6 (Sβ=0.074, P=0.009), IL-8 (Sβ=0.089, P=0.002), TNF-ɑ (Sβ=0.0.082, P=0.005), E-selectin (Sβ=0.071, P=0.016) and intercellular adhesion molecule-1 (ICAM-1, Sβ=0.117, P<0.001) after adjustments were made for age, gender, race, marital status, education, current smoking status, physician-diagnosed CVDs and respiratory diseases, BMI and perceived stress. However, the global sleep score was not associated with serum IL-10 (Sβ=−0.021, P=0.463) and CRP (Sβ=0.035, P=0.059) levels after adjustments were made for these confounding factors.ConclusionPoor sleep is positively associated with serum inflammatory marker levels among obese adults. Sufficient sleep may be particularly important for obese adults to prevent CVDs.

Ropivacaine via nuclear factor kappa B signalling modulates CD62E expression and diminishes tumour cell arrest.

The issue whether anaesthesia has an impact on the prognosis of carcinoma has been widely discussed and remains debated. Ropivacaine has been widely used in perioperative period as a long acting local anesthetic. An early event during recurrence or metastasis of carcinoma is the adhesion of circulating tumour cells (CTCs) to endothelial cells (ECs) through binding adhesion molecules that are up-regulated on inflamed endothelium during the perioperative period or other periods. This study was to explore the impact of ropivacaine on the adhesion of tumour cells, providing evidences of its influence on the prognosis of carcinoma. Human umbilical vein endothelial cells (HUVECs) were pre-treated with ropivacaine (10-7-10-5 M; 30 min) prior to treatment with tumour necrosis factor alpha (TNFα) (10 ngml-1; 1, 4 and 8 h). Intercellular adhesion molecule-1 (ICAM-1), endothelial-selectin (CD62E) and vascular cell adhesion molecule-1 (VCAM-1) mRNA levels were detected via quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). To clarify the underlying action mechanism, p65, p-p65, IκBα, p-IκBα, IKKα/β and p-IKKα/β protein levels were evaluated via western blotting. Cell viability and tumour cell adhesion assays were also assessed. The clinically usage concentration of ropivacaine (10-6 M) produced a significant decrease in CD62E expression compared with that produced by TNFα only (p < 0.001). Moreover, adhesion assays showed that ropivacaine effectively inhibited the adhesion of hepatoma cells (p < 0.01), human colon cancer cells (p < 0.01) and human leukemic monocyte (p < 0.01). Western blot results showed that pre-treatment with ropivacaine inhibited the phosphorylation of p65 (p < 0.05), IκBα (p < 0.001) and IKKα/β (p < 0.01). Ropivacaine decreased the adhesion of tumour cells. Ropivacaine modulated CD62E expression by inhibiting the activation of NF-κB. These results might provide new insight into the issue whether anaesthesia has an impact on the prognosis of carcinoma.

MicroRNA-29a-3p Reduces TNFα-Induced Endothelial Dysfunction by Targeting Tumor Necrosis Factor Receptor 1

miR-29a-3p has been shown to be associated with cardiovascular diseases; however, the effect of miR-29a-3p on endothelial dysfunction is unclear. This study aimed to reveal the effects and mechanisms of miR-29a-3p on endothelial dysfunction. The levels of vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), and E-selectin were determined by real-time PCR and immunofluorescence staining to reveal the degree of tumor necrosis factor alpha (TNFα)-induced endothelial dysfunction. A luciferase activity assay and cell transfection with a miR-29a-3p mimic or an inhibitor were used to reveal the underlying mechanisms of miR-29a-3p action. Furthermore, the effects of miR-29a-3p on endothelial dysfunction were assessed in C57BL/6 mice injected with TNFα and/or a miR-29a-3p agomir. The results showed that the expression of TNFα-induced adhesion molecules in vascular endothelial cells (EA.hy926 cells, human aortic endothelial cells [HAECs], and primary human umbilical vein endothelial cells [pHUVECs]) and smooth muscle cells (human umbilical vein smooth muscle cells [HUVSMCs]) was significantly decreased following transfection with miR-29a-3p. This effect was reversed by cotransfection with a miR-29a-3p inhibitor. As a key target of miR-29a-3p, tumor necrosis factor receptor 1 mediated the effect of miR-29a-3p. Moreover, miR-29a-3p decreased the plasma levels of TNFα-induced VCAM-1 (32.62%), ICAM-1 (38.22%), and E-selectin (39.32%) in vivo. These data indicate that miR-29a-3p plays a protective role in TNFα-induced endothelial dysfunction, suggesting that miR-29a-3p is a novel target for the prevention and treatment of atherosclerosis.

Intercellular adhesion molecule-1 expression and serum levels as markers of pre-clinical atherosclerosis in polycystic ovary syndrome

BackgroundPolycystic ovary syndrome (PCOS) is a common reproductive endocrine disorder characterized by obesity, hyperandrogenism, and insulin resistance. Intercellular adhesion molecule-1 (ICAM-1) is a proinflammatory and proatherogenic cytokine which is associated with atherosclerosis, insulin resistance, and cardiovascular disease (CVD). The pathogenesis of PCOS is not precisely known. Thus, the purpose of this study was to investigate the potential role of ICAM-1 expression and serum ICAM-1 concentrations in pathogenesis of PCOS. Moreover, we aimed to evaluate the possible relationship between ICAM-1 gene expression with carotid intima-media thickness as well as clinic-morphological features of PCOS.MethodsThis case control study enrolled 180 patients with PCOS and 120 controls groups and they were stratified according to their fasting plasma glucose (FPG) into three subgroups; normal glucose tolerance (NGT) [n = 75], those with impaired glucose tolerance (IGT) [n = 65], and 40 patients with type 2 diabetes mellitus (T2DM). Circulating ICAM-1 expression levels were determined by real time polymerase chain reaction (RT-PCR). Serum ICAM-1 concentrations were measured using enzyme-linked immunosorbent assay (ELISA).ResultsOur results revealed that PCOS patients had higher values of ICAM-1expression and serum levels. Among PCOS patients, T2DM patients had the highest values of ICAM-1 expression and serum levels compared to IGT and NGT subgroups. The ICAM-1 expression and serum levels were significantly positive correlated with cardiovascular risk and PCOS phenotypes. Linear regression test showed that HOMA-IR was the main predictors of serum ICAM-1 levels in PCOS. Receiver operating characteristic curve (ROC) analysis revealed that, the power of ICAM-1 expression levels was higher than serum ICAM-1 in diagnosis of PCOS and in differentiating T2DM from IGT and NGT subgroups. Interestingly, combination of both ICAM-1 expression and serum levels improved the diagnostic role of serum ICAM-1.ConclusionICAM-1 expression and serum levels were higher in women with PCOS compared to control group also, there was a strong independent association between higher ICAM-1 expression and serum levels with cardiovascular risks in PCOS group.

Screening and identification of potential active components in Ophiopogonis Radix against atherosclerosis by biospecific cell extraction

Atherosclerosis is a chronic disease and an important pathological process associated with cardiovascular disease. Endothelial dysfunction, vascular smooth muscle cells (VSMCs) proliferation and neutrophil activation are involved in the development of atherosclerosis. Ophiopogonis Radix is a common traditional Chinese medicine use to treat cardiovascular diseases, however, its active constituents remain to be elucidated. In this study, primary vascular endothelial cells, primary VSMCs and neutrophils were prepared, and extract of Ophiopogonis Radix (EOR) was investigated to ameliorate H2O2 induced reactive oxygen species (ROS) and nitric oxide (NO) production. The results showed that EOR decreased levels of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1, its protective effects against oxidative damage of endothelia and endothelial dysfunction. Additionally, EOR treatment inhibited oxidized low-density lipoprotein-induced VSMC proliferation, phorbol-12-myristate-13-acetate-mediated ROS production and neutrophil activation, malondialdehyde production, and decreased superoxide dismutase activity and myeloperoxidase release. By HPLC-Q-TOF-MS/MS analysis, 51 compounds in EOR were identified including 22 saponins and 24 homoisoflavonoids. Then biospecific cell extraction and LC-MS technique were employed to screening the antiatherosclerosis active components in Ophiopogonis Radix. After co-cultured with EOR, the multi-effective active constituents including four saponins and two homoisoflavonoids were acquired and subsequently verified to restore properties including endothelial injury, VSMC proliferation and neutrophil activation, indicating that these compounds may be multi-effective active constituents that were responsible for atherosclerosis and the cardiovascular protection of Ophiopogonis Radix.

P5343The presence of anti-apolipoprotein A1 autoantibodies is associated with a pro-atherogenic profile in HIV-infected patients

Abstract Background With the access to antiretroviral therapy (ART), the mortality related to the human immunodeficiency virus (HIV) has dropped, shifting the clinical challenges towards chronic disease management, including cardiovascular disease (CVD) risk assessment. Factors that potentially contribute to the physiopathology of HIV-related CVD include the HI-virus itself, adverse effects of ART, and processes such as dyslipidemia, inflammation, immune/autoimmune activation and endothelial injury. Among autoantibodies of possible cardiovascular relevance, those directed against apolipoprotein A-1 (anti-apoA-1 IgG) were shown to predict major adverse cardiovascular events and promote atherogenesis. Purpose The aim of the study was to evaluate the prevalence of anti-apoA1 IgG in HIV-free and ART experienced and naïve HIV-infected patients as well as the association between anti-apoA1 IgG levels and, indices of viral suppression, clinical parameters (10 year Framingham Risk Score (FRS)) and inflammatory biomarkers, known to underlie atherosclerosis burden in these patients. Methods Anti-apoA1 IgG serum levels were assessed by a homemade ELISA assay in 144 participants from a South African cohort divided in three groups: HIV-free (n=50), HIV-infected/ART experienced (n=50) and HIV-infected/ART naïve (n=44). Inflammatory biomarkers were measured. Results HIV-infected patients displayed an increased pro-atherogenic biomarker profile compared to HIV-free subjects, but not difference in the FRS was observed between these two groups. Regarding anti-apoA1 IgG, 24% of HIV-free patients tested positive compared to 40% and 70% in HIV-infected/ART experienced and naïve groups, respectively. HIV-infected, anti-apoA1 IgG positive patients showed a significant decrease in CD4+ counts (p=0.003) and a significant increase in viremia (p=0.0130), mean heart rate (p=0.0243), albuminuria (p=0.0155), pro-inflammatory biomarkers (IFNγ, IL-10, TNFα, MIPα; all p&lt;0.05), circulating levels of intercellular adhesion molecule (ICAM-1) (p=0.0217) and vascular cell adhesion molecule (VCAM-1) (p=0.003) compared to anti-apoA1 IgG negative ones. Of note, while this profile was maintained in HIV-infected/ART experienced, these significant differences were lost in HIV-infected/ART naïve patients. No significant difference in FRS was observed between anti-apoA1 IgG positive vs negative individuals in all groups. Conclusions HIV-infected patients presented with an increased prevalence of anti-apoA1 IgG compared to HIV-free subjects. In HIV-infected/ART experienced patients, anti-apoA1 IgG levels were associated with low CD4+ counts, levels of adhesion molecules and pro-inflammatory responses, features associated with increased cardiovascular events. ART highlighted pro-atherogenic differences between HIV-infected anti-apoA1 IgG negative and positive patients.

P6289Neuregulin-1 improves cardiac function through protecting the loss of endothelial barrier in septic rats via RhoA/ROCK pathway

Abstract Background The main path of sepsis-induced cardiomyopathy is the loss of endothelial barrier function. Neuregulin-1 (NRG-1) has beneficial effects on endothelial function. Purpose To investigate the effect of NRG-1 treatment on the protection of cardiac endothelial cells and the changes of RhoA/ROCK signaling in sepsis. Methods Rats were randomly divided into three groups: sham, LPS, and NRG-1. After successful induction of sepsis by lipopolysaccharide (LPS, 10mg/kg), rats were administered either a vehicle or recombinant human neuregulin-1 (rhNRG-1, 10μg/kg/d) for one or two days. We recorded their survival rate at 48h after sepsis. Hemodynamic methods were performed to assess cardiac function. We used immunofluorescence assay to detect von Willebrand Factor (vWF). Intercellular adhesion molecule-1 (ICAM-1) and vascular endothelial growth factor (VEGF)levels in serum were measured by enzyme-linked immunosorbent assay (ELISA), and serum nitric oxide (NO) was detected by reductase method. We used transmission electron microscopy to observe changes in myocardial ultrastructure and western blot to assess expression of RhoA and ROCK1 protein. Results Sepsis impaired endothelial function manifested by increased ICAM-1, NO and VEGF levels in serum, NRG-1 treatment could significantly alleviate these increase (P&lt;0.05). Compared with the vehicle, NRG-1 significantly decreased cardiac vascular permeability through increasing the expression of vWF on endothelial lining (P&lt;0.05). Moreover, NRG-1 alleviated damages of ultrastructure of myocardial cells and suppressed the expression of RhoA and ROCK1 protein (P&lt;0.05). Ultimately, NRG-1 improved the survival,and prevented hemodynamic derangement. Levels of endothelial biomarker in serum ICAM-1 (pg/ml) VEGF (pg/ml) NO (pg/ml) Sham 24h 1564.74±94.41 0.84±0.28 203.27±1.81 Sham 48h 322.92±7.92 1.78±0.61 6.57±0.38 LPS 24h 5139.85±284.15† 3.11±0.04† 679.05±78.59† LPS 48h 950.41±25.23†‡ 4.65±0.30† 180.40±30.08†‡ NRG 24h 2772.18±164.45§ 0.30±0.04§ 355.14±23.41§ NRG 48h 578.57±28.97§‡ 1.02±0.45§ 104.67±2.13§‡ †p&lt;0.05 vs. the sham group, §p&lt;0.05 vs. the LPS group and ‡p&lt;0.05 vs. the 24h group. Effect of NRG-1 on endothelial cells Conclusions NRG-1 could alleviate endothelial injury in sepsis by strengthening the barrier function of vascular, reducing the secretion of endothelial-related biomarkers and inhibiting oxidative stress, thus improving cardiac function and survival rate, these effects may base on RhoA/ROCK signaling pathway. These may contribute to reverse the impaired endothelial cells in sepsis in the future. Acknowledgement/Funding The national natural science foundation of China (81772044)

Biomarkers of Systemic Lupus Erythematosus and Systemic Sclerosis diseases activity in a sample of Egyptian patients :Soluble Intercellular Adhesion Molecule-1 and Soluble Interleukin-2 Receptor, Case Control Study

Systemic Lupus Erythematosus (SLE) and systemic sclerosis (SSc) are systemic inflammatory autoimmune disorders characterized by a large spectrum of clinical and laboratory features. The aim of the present study was to investigate the possible use of serum level of soluble intercellular adhesion molecule-1(sICAM-1) and soluble interleukin-2 receptor (sIL-2Ra) as biomarkers for monitoring of SLE and SSc disease activity. Moreover, it aimed to compare the specificity and sensitivity as well as cut-off value of both biomarkers in a sample of Egyptian patients. 50 SLE patients, 30 SSc patients and 60 age and sex matched healthy controls were enrolled in our study. sICAM-1and sIL-2Ra were measured in serum samples obtained from all participants. In addition to Erythosedimentation rate (ESR), complete blood count (CBC), Antineuclearantibodies (ANA) estimation, disease activity of both diseases were also assessed. sICAM-1and sIL-2Ra levels were higher in SLE and SSc patients versus control. Both parameters are correlated with each other as well as the activity parameters. A cut-off levels of 455.59 (ng/ml) &amp;2525935 (pg/ml) in both SLE &amp; SSs respectively was observed with the highest specificity and sensitivity. It could be concluded that sICAM-1 and sIL-2Ra are noninvasive biomarkers for SLE and SSc that could play a pathophysiologic role in development and progression of both diseases. Moreover, sICAM-1 and sIL-2Ra are correlated with the disease activity at cut-off values of 455.59 (ng/ml) &amp; 2525935(pg/ml) respectively.

Oncolytic Immunotherapy for Bladder Cancer Using Coxsackie A21 Virus: Using a Bladder Tumor Precision-Cut Slice Model System to Assess Viral Efficacy.

Oncolytic viruses are anticancer agents that selectively target and kill cancer cells by direct lysis, while at the same time stimulating a tumor antigen-specific adaptive immune response. These promising therapeutic agents target multiple cancers and have already proven to be an effective treatment option for solid malignancies. One such agent, T-Vec (Talimogene laherparepvec) has been licensed and is in routine clinical use for treatment of malignant melanoma.Non-muscle invasive bladder cancer (NMIBC) is an ideal potential target for oncolytic immunotherapy as locally instilled live biological therapy using Bacille Calmette-Guerin (BCG) is already well established in the clinical setting. Coxsackievirus A21 (CVA21) is a novel intercellular adhesion molecule-1 (ICAM-1)-targeted immunotherapeutic virus. We have investigated CVA21-induced cytotoxicity in a panel of human bladder cancer cell lines, revealing a range of sensitivities largely correlating with expression of the viral receptor ICAM-1. CVA21 in combination with low doses of mitomycin-C enhanced CVA21 viral replication and oncolysis by increasing surface expression levels of ICAM-1. In addition to cell lines and an animal model a key component of our studies into oncolytic immunotherapy for bladder cancer was the use of a bladder tumor precision slice preclinical model system which represents tumor architecture, heterogeneity, and the complexity of a tumor in vitro. Results seen in cell lines were reflected in the tumor slice model whereby levels of virus protein expression and induction of apoptosis were enhanced with prior exposure to mitomycin-C. In this chapter we demonstrate the utility of the precision cut tumor slice model as a unique organotypic model to test oncolytic viruses. We will describe how to prepare and slice the tumor using a vibrating microtome together with the optimum culture and conditions for treatment.