Intracellular Adhesion Molecule-1 Levels Research Articles

Endotoxin-induced and vaccine-induced systemic inflammation both impair endothelium-dependent vasodilation, but not pulse wave reflection

BackgroundInflammation induced by either endotoxin or vaccination has previously been shown to impair endothelium-dependent vasodilation (EDV) in healthy young individuals. However, the vascular effects of these two mechanisms of inducing inflammation have not been compared in the same individuals.MethodsTwelve young healthy males were studied at the same time of the day on three occasions in a random order; on one occasion 4 hours following an endotoxin injection (Escherichia coli endotoxin, 20 IU/kg), on another occasion 8 hours following vaccination against Salmonella typhi, and on a third occasion 4 hours following a saline control injection. EDV and endothelium-independent vasodilation (EIDV) were evaluated by local infusions of acetylcholine and sodium nitroprusside in the brachial artery, and forearm blood flow was measured with venous occlusion plethysmography. The augmentation index was determined by pulse wave analysis as an index of pulse wave reflection.ResultsBoth endotoxin and vaccination impaired EDV to a similar degree compared with the saline control (P = 0.005 and P = 0.014, respectively). EIDV was not significantly affected by inflammation. Endotoxin, but not vaccination, increased body temperature and circulating levels of intracellular adhesion molecule-1 and interleukin-6. Augmentation index was not affected by the interventions.ConclusionDespite the fact that endotoxin induced a more pronounced degree of inflammation than vaccination, both inflammatory challenges impaired EDV to a similar degree, supporting the view that different inflammatory stimuli could induce harmful effects on the vasculature.

Protective effects of kaerophyllin against liver fibrogenesis in rats

We previously demonstrated that kaerophyllin, a lignan, isolated from a widely used traditional Chinese herb, Bupleurum scorzonerifolium, leading to the inhibition of hepatic stellate cells (HSCs) activation in vitro. This current study evaluated the in vivo role of kaerophyllin in protecting the liver against injury and fibrogenesis caused by thioacetamide (TAA) in rats and further explored the underlying mechanisms. Liver fibrosis in Sprague-Dawley rats was induced by intraperitoneal injection of TAA (200 mg/kg) twice per week for 6 weeks. Animals were divided into five groups: vehicle control, TAA control, TAA + low dose kaerophyllin, TAA + high dose kaerophyllin and TAA + curcumin groups. Kaerophyllin (10 or 30 mg/kg) or curcumin (150 mg/mL) was given by gavage twice per day consecutively for 4 weeks starting 2 weeks after TAA injection. Rat HSCs were used to investigate the anti-inflammatory role of kaerophyllin against tumour necrosis factor α (TNF-α) in vitro. Peroxisome proliferator-activated receptor-γ (PPAR-γ) expression was knocked down in rat HSCs using PPAR-γ small interfering RNAs. Kaerophyllin significantly protected liver from injury by reducing serum aspartate transaminase and alanine transaminase levels and by improving the histological architecture and fibrosis score. In addition, kaerophyllin suppressed inflammation by reducing the mRNA of TNF-α, interleukin-1β (IL-1β) and monocyte chemoattractant protein-1 (MCP-1) genes. In HSCs, kaerophyllin elevated PPAR-γ activity and reduced TNF-α-stimulated mRNA levels of intracellular adhesion molecule-1 (ICAM-1), MCP-1 and IL-1β genes, which were reversed by small interfering RNA knockdown of PPAR-γ gene. Our results demonstrated that kaerophyllin protected the rat liver from TAA-caused injury and fibrogenesis by suppressing hepatic inflammation and inhibiting HSC activation, possibly through upregulation of PPAR-γ expression.

Flavonoids inhibit high glucose-induced up-regulation of ICAM-1 via the p38 MAPK pathway in human vein endothelial cells

Recently, several flavonoids have been shown to have cardioprotective, cancer preventive, or anti-inflammatory properties. However, the specific mechanisms underlying their protective effects remain unclear. We aimed to investigate the different effects of three representative flavonoids—hesperidin, naringin, and resveratrol—on intracellular adhesion molecule-1 (ICAM-1) induction in human umbilical vein endothelial cells (HUVECs) by using high-glucose (HG) concentrations and the possible underlying molecular mechanisms. In HG-induced HUVEC cultures, the effects of three different flavonoids on ICAM-1 production and p38 phosphorylation were examined in the presence or absence of inhibitors targeting the mitogen-activated protein kinase (MAPK) signal transduction pathway. HG stimulation of HUVECs increased the levels of the adhesion molecules ICAM-1 and endothelial selectin (E-selectin). Pretreatment with all the three flavonoids drastically inhibited ICAM-1 expression in a time-dependent manner, but did not alter VCAM-1 and E-selectin expressions. Moreover, we investigated the effects of flavonoids on the MAPK signal transduction pathway, because MAPK families are associated with vascular inflammation under stress. These flavonoids did not block HG-induced phosphorylation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), but completely inhibited the HG-induced phosphorylation of p38 MAPK. SB202190, an inhibitor of p38 MAPK, also inhibited the HG-induced enrichment of ICAM-1. This study demonstrated that hesperidin, naringin, and resveratrol reduced the HG-induced ICAM-1 expression via the p38 MAPK signaling pathway, contributing to the inhibition of monocyte adhesion to endothelial cells.

Statin therapy attenuates growth and malignant potential of human esophageal adenocarcinoma cells

Esophageal adenocarcinoma is an aggressive malignancy generally diagnosed after metastatic spread and currently lacks effective medical therapy. Expression of intracellular adhesion molecule-1 (ICAM-1) is an adverse prognostic indicator in various human tumor cells and contributes significantly to their metastatic potential. Statin therapy reduces circulating ICAM-1 levels in patients with coronary heart disease and is associated with reduction in progression from Barrett esophagus to esophageal adenocarcinoma. We hypothesize that statin therapy may attenuate growth and malignant potential via ICAM-1 expression and nuclear factor-kappa beta activation in human esophageal adenocarcinoma cells. Verified human esophageal adenocarcinoma cells (FLO-1) were treated with simvastatin, atorvastatin, or pravastatin (10-, 30-, and 50-μmol/L concentrations). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide viability, 5-bromo-2'-deoxyuridine proliferation, or annexin V apoptosis assays were performed, or cells were stimulated with tumor necrosis factor-alpha and collected for immunoblotting and flow cytometry. Simvastatin decreased cell viability and proliferation while increasing apoptosis in a dose-dependent manner (P < .05). Simvastatin attenuated total cellular and cell-surface ICAM-1 expression as well as nuclear factor-kappa beta activation (P < .05). Atorvastatin had mild effects and pravastatin had essentially no effect on growth and metastatic potential of these cells. We demonstrate that treatment of human esophageal adenocarcinoma cells with simvastatin attenuates growth, by decreasing cell viability, decreasing cell proliferation, and increasing apoptosis, and attenuates metastatic potential, by decreasing expression of key metastatic markers. These findings identify simvastatin as a potential therapeutic and chemopreventive modality to thwart the progression of esophageal adenocarcinoma.

Piper sarmentosum inhibits ICAM-1 and Nox4 gene expression in oxidative stress-induced human umbilical vein endothelial cells

BackgroundAqueous extract of Piper sarmentosum (AEPS) is known to possess antioxidant and anti-atherosclerotic activities but the mechanism responsible for it remains unclear. In early part of atherosclerosis, nuclear factor-kappa B (NF-κB) induces the expression of cellular adhesion molecules such as vascular cell adhesion molecule-1 (VCAM-1), intracellular adhesion molecule-1 (ICAM-1) and E-selectin. NADPH oxidase 4 (Nox4) is the predominant source of superoxide in the endothelial cells whereas superoxide dismutase 1 (SOD1), catalase (CAT) and glutathione peroxidase (GPx) are the antioxidant enzymes responsible for inactivating reactive oxygen species. The present study aimed to investigate the effects of AEPS on the gene expression of NF-κB, VCAM-1, ICAM-1, E-selectin, Nox4, SOD1, CAT and GPx in cultured human umbilical vein endothelial cells (HUVECs).MethodsHUVECs were divided into four groups:- control; treatment with 180 μM hydrogen peroxide (H2O2); treatment with 150 μg/mL AEPS and concomitant treatment with AEPS and H2O2 for 24 hours. Total RNA was extracted from all the groups of HUVEC using TRI reagent. Subsequently, qPCR was carried out to determine the mRNA expression of NF-κB, VCAM-1, ICAM-1, E-selectin, Nox4, SOD1, CAT and GPx. The specificity of the reactions was verified using melting curve analysis and agarose gel electrophoresis.ResultsWhen stimulated with H2O2, HUVECs expressed higher level of ICAM-1 (1.3-fold) and Nox4 (1.2-fold) mRNA expression. However, AEPS treatment led to a reduction in the mRNA expression of ICAM-1 (p < 0.01) and Nox4 (p < 0.05) in the H2O2-induced HUVECs. AEPS also upregulated the mRNA expression of SOD1 (p < 0.05), CAT (p < 0.01) and GPx (p < 0.05) in oxidative stress-induced HUVECs. There was no significant change in the mRNA expression of VCAM-1 and E-selectin.ConclusionThe expressional suppression of ICAM-1 and Nox4 and induction of antioxidant enzymes might be an important component of the vascular protective effect of AEPS.

ASSOCIATION BETWEEN NOVEL BIOMARKERS AND ARTERIAL ELASTICITY INDEX AMONG HEALTHY MULTI-ETHNIC U.S. POPULATION

Methods: Multi-Ethnic study of Atherosclerosis (MESA) is a population based study (n=6,814) of White (38%), Black (28%), Chinese (22%) and Hispanic (12%) subjects, aged 45-84 yrs, free from clinical cardiovascular disease. We performed post-hoc analysis of the NHLBI limited access dataset of MESA subjects (n=6,002) using multivariate regression analyses to determine the association of biomarkers with LAE and SAE derived from radial artery diastolic pulse wave contour registration in all subjects in a supine position using tonometry. Results: C reactive protein, interleukin-6, fibrinogen and factor-8 were negatively associated with both LAE and SAE. Plasma activated protein, homocysteine and microalbuminuria were negatively associated with SAE only and intracellular adhesion molecule levels were associated negatively with LAE. Conclusion: Markers of inflammation are associated with both LAE and SAE, whereas, markers of endothelial dysfunction and hemostasis are more strongly associated with SAE. The differential association of these biomarkers with LAE and SAE may further help our understanding of the putative biological pathways mediating adverse cardiovascular outcomes. Table 1: Association of Novel biomarkers with Arterial Elasticity Index Small Arterial Elasticity Index Large Arterial Elasticity Index Biomarkers Beta-coefficient [95% CI] p-value Beta-coefficient [95% CI] p-value High-sensitivity C-reactive protein (mg/L) -0.015 [-0.023 to -0.007] <0.001 -0.029 [-0.041 to -0.017] <0.001 Interleukin-6 (pg/mL) -0.030 [-0.043 to -0.016] <0.001 -0.036 [-0.057 to -0.016] <0.001 Plasmin-Antiplasmin Complex (nM) -0.041 [-0.064 to -0.017] 0.001 -0.012 [-0.022 to -0.047] 0.483 Fibrinogen (mg/dL) -0.071 [-0.115 to -0.027] 0.002 -0.086 [-0.150 to -0.020] 0.008 D-dimer (mcg/mL) -0.001 [-0.011 to 0.009] 0.864 0.009 [-0.005 to 0.023] 0.215 Factor-8 (%) -0.029 [-0.050 to -0.008] 0.005 -0.030 [-0.060 to -0.001] 0.049 Homocysteine (micromol/L) -0.037 [-0.067 to -0.006] 0.017 -0.032 [-0.012 to 0.076] 0.155 *Microalbuminuria (mg/g of creatinine) -0.039 [-0.071 to -0.007] 0.015 -0.015 [-0.061 to 0.031] 0.530 ¶Intercellular adhesion molecule-1 (ng/ml) -0.033 [-0.083 to 0.016] 0.188 -0.144 [-0.216 to -0.072] <0.001

Levocetrizine has anti-inflammatory effects against Toll-like receptor (TLR)3 through the inhibition of Toll-IL-1 receptor (TIR)-domain-containing adapter inducing IFN-beta (TRIF) and receptor-interacting protein (RIP)

To investigate the anti-inflammatory effect of levocetrizine (LCEZ) on the intracellular adhesion molecule-1 (ICAM-1) in human nasal epithelial cells stimulated by TLR3 and further analyze the anti-inflammatory mechanism of LCEZ in the MyD88-independent pathway before NF-κB is activated. A primary culture of human nasal epithelial cells (HNECs) was generated from nasal polyps. After stimulation of epithelial cells with LTA, double-stranded RNA (dsRNA), and LPS, reverse transcription-polymerase chain reaction (RT-PCR) was performed at 1, 6, and 24 h to clarify the optimal stimulation of ICAM-1 in HNECs. To investigate the anti-inflammatory effects of LCEZ, HNECs were pretreated with three different concentrations of LCEZ (500, 50, and 5 nM) for 2 h. HNECs were washed and then stimulated with dsRNA. At 1, 6, and 24 h after stimulation, the level of ICAM-1 was measured by RT-PCR and ELISA. Western blots for TRIF and RIP were performed. The level of ICAM-1 was significantly elevated by dsRNA. Pretreatment with LCEZ decreased the secretion of ICAM-1, which was observed in RT-PCR and Western blots but not in ELISA analyses. The expression of TRIF and RIP, measured by Western blot, was decreased by pretreatment with LCEZ. The activation of HNECs by TLRs (especially TLR3) could trigger an inflammatory process, which might be inhibited by LCEZ through the suppression of TRIF and RIP proteins.

Evaluation of Morroniside, Iridoid Glycoside from Corni Fructus, on Diabetes-Induced Alterations such as Oxidative Stress, Inflammation, and Apoptosis in the Liver of Type 2 Diabetic db/db Mice

The present study was conducted to examine whether morroniside has an ameliorative effect on diabetes-induced alterations such as oxidative stress, inflammation, and apoptosis in the liver of type 2 diabetic db/db mice. Morroniside (20 or 100 mg/kg body weight/d, per os (p.o.)) was administered every day for 8 weeks to db/db mice, and its effect was compared with vehicle-treated db/db and m/m mice. The administration of morroniside decreased the elevated serum glucose concentration in db/db mice, and reduced the increased oxidative biomarkers including the generation of reactive oxygen species and lipid peroxidation in the liver. The db/db mice exhibited the up-regulation of nicotinamide adenine dinucleotide phosphate oxidase subunits, NF-E2-related factor 2 (Nrf2), heme oxygenase-1, nuclear factor-kappa B, cyclooxygenase-2, inducible nitric oxide synthase, monocyte chemotactic protein-1, and intracellular adhesion molecule-1 levels in the liver; however, morroniside treatment significantly reduced those expressions. Moreover, the augmented expressions of apoptosis-related proteins, Bax and cytochrome c, were down-regulated by morroniside administration. Hematoxylin-eosin staining showed that the increased hepatocellular damage in the liver of db/db mice improved on morroniside administration. Taking these into consideration, our findings support the therapeutic evidence for morroniside ameliorating the development of diabetic hepatic complications via regulating oxidative stress, inflammation, and apoptosis.

Inflammatory Response in the First 48 Hours of Acute Ischemic Stroke

Background : Activation of endothelial cells is an important mediator of atherothrombosis. Markers of endothelial cells such as soluble adhesion molecules can be measured in plasma and reflect the activity of the endothelium and the inflammatory system. We hypothesized that patients with acute ischemic stroke would have a dynamic change in their markers of inflammation over time, primarily reflecting the activation of endothelial cells and the immunological ability to respond to an acute brain insult. We also believed that the acute inflammatory response in the first 72 hours might affect the short and long term clinical outcome. Methods : We conducted a prospective case study of 27 patients that were admitted with acute ischemic stroke during the years 2005 - 2007. All were examined clinically using the National Institute of Health Stroke Scale [NIHSS] and a brain computed tomography (CT) scan was done in the first 24 hours. Blood was drawn for levels of E-selectin, intracellular adhesion molecule 1 (ICAM-1), and vascular cellular adhesion molecule 1 (VCAM-1) on admission and 48 hours later by ELISA methods. The blood was separated and the serum was frozen at -80 ° C until analyzed as one batch. Results : Mean blood concentrations of soluble E-selectin, intracellular adhesion molecule 1 (ICAM-1), and vascular cell adhesion molecule 1 (VCAM-1) were measured on admission and 48 hours later. Clinically there were 3 groups: 6 patients with transient ischemic attack [TIA] (58 ± 12 years old, 3 women and 3 men), 8 patients with cerebrovascular accident [CVA] without recovery (75 ± 18 years old, 4 women and 4 men), and 13 patients with CVA who recovered clinically (70 ± 13 years old, 6 women and 7 men). There was a significant increase in E-selectin levels in the second measurement (from 27.5 ± 21.6 ng/ml to 38.7 ± 19.6 ng/ml; Z = -1.997, P = 0.046) in the TIA group. An inverse correlation was found between E-selectin level and age among TIA patients on admission (r = -0.913, P = 0.011) and 48 hours later (r = -0.850, P = 0.032). A positive correlation between ICAM-1 and VCAM-1 levels was found 48 hours post admission (r = 0.436, P = 0.026). Conclusions : We have demonstrated a significant increase in E-selectin level within 48 hours among patients with TIA. In the TIA group there was an inverse correlation between age and E-selectin level. This may suggest that younger patients can protect their ischemic brain more efficiently due to a more competent immune system, and that the immune system may have an important role in ischemic brain injury. J Neurol Res. 2011;1(1):16-21 doi: https://doi.org/10.4021/jnr101e

Inflammatory Response in the First 48 Hours of Acute Ischemic Stroke

Background : Activation of endothelial cells is an important mediator of atherothrombosis. Markers of endothelial cells such as soluble adhesion molecules can be measured in plasma and reflect the activity of the endothelium and the inflammatory system. We hypothesized that patients with acute ischemic stroke would have a dynamic change in their markers of inflammation over time, primarily reflecting the activation of endothelial cells and the immunological ability to respond to an acute brain insult. We also believed that the acute inflammatory response in the first 72 hours might affect the short and long term clinical outcome. Methods : We conducted a prospective case study of 27 patients that were admitted with acute ischemic stroke during the years 2005 - 2007. All were examined clinically using the National Institute of Health Stroke Scale [NIHSS] and a brain computed tomography (CT) scan was done in the first 24 hours. Blood was drawn for levels of E-selectin, intracellular adhesion molecule 1 (ICAM-1), and vascular cellular adhesion molecule 1 (VCAM-1) on admission and 48 hours later by ELISA methods. The blood was separated and the serum was frozen at -80 ° C until analyzed as one batch. Results : Mean blood concentrations of soluble E-selectin, intracellular adhesion molecule 1 (ICAM-1), and vascular cell adhesion molecule 1 (VCAM-1) were measured on admission and 48 hours later. Clinically there were 3 groups: 6 patients with transient ischemic attack [TIA] (58 ± 12 years old, 3 women and 3 men), 8 patients with cerebrovascular accident [CVA] without recovery (75 ± 18 years old, 4 women and 4 men), and 13 patients with CVA who recovered clinically (70 ± 13 years old, 6 women and 7 men). There was a significant increase in E-selectin levels in the second measurement (from 27.5 ± 21.6 ng/ml to 38.7 ± 19.6 ng/ml; Z = -1.997, P = 0.046) in the TIA group. An inverse correlation was found between E-selectin level and age among TIA patients on admission (r = -0.913, P = 0.011) and 48 hours later (r = -0.850, P = 0.032). A positive correlation between ICAM-1 and VCAM-1 levels was found 48 hours post admission (r = 0.436, P = 0.026). Conclusions : We have demonstrated a significant increase in E-selectin level within 48 hours among patients with TIA. In the TIA group there was an inverse correlation between age and E-selectin level. This may suggest that younger patients can protect their ischemic brain more efficiently due to a more competent immune system, and that the immune system may have an important role in ischemic brain injury. doi:10.4021/jnr101e

Abstract P2-16-04: Plasma Biomarker Analyses in the AVADO Phase III Randomized Study of First-Line Bevacizumab + Docetaxel in Patients with Human Epidermal Growth Factor Receptor (HER) 2-Negative Metastatic Breast Cancer

Abstract Background: Inhibition of vascular endothelial growth factor (VEGF) signaling by bevacizumab (Bv; Avastin) combined with chemotherapy (chemo) is associated with improved clinical benefit in several malignancies, including metastatic breast cancer (mBC). Bv+chemo was evaluated in HER2 negative mBC in AVADO where baseline biomarker (BM) sampling was included as an optional element to identify prognostic and predictive biomarkers. Different angiogenic markers have been evaluated in trials containing Bv, with most trials having focused on circulating VEGF. Thus far, VEGF seems to have more a prognostic than predictive value as demonstrated across different indications (lung, renal, and colorectal cancer [Bernaards et al. ASCO 2010]). Trends for the predictive value of intracellular adhesion molecule-1 (ICAM-1) have been noted in two NSCLC studies (E4599 and AVAiL) and for basic fibroblast growth factor in AVAiL. The value of angiogenic plasma markers has not yet been established in mBC. Material and Methods: In AVADO, 736 mBC patients (pts) were randomized to docetaxel 100mg/m2 plus either Bv7.5mg/kg, Bv15mg/kg or placebo. Primary endpoint was progression-free survival (PFS). Baseline plasma samples were analyzed for VEGF, ICAM-1, VEGF Receptor 1 and 2 (VEGFR1, 2), E-selectin and vascular cell adhesion molecule-1 using a novel ELISA based multiplex assay. Samples for BM analysis were available for 396 pts, equally distributed across treatment arms. Median levels of these markers were pre-specified as a cut point to categorize pts as low or high. Pre-treatment values were correlated with PFS (before start of subsequent antineoplastic therapy) using simple and multiple regression approaches as well as subgroup analyses. Results: All subgroups had HR point estimates in favor of the Bv arms, with the exception of pts with low VEGFR2 levels at a Bv dose of 7.5mg/kg. In general, high plasma levels of VEGF and VEGFR2 were associated with better treatment effect in this study. Low ICAM-1 levels did not show the trend towards a larger treatment effect previously seen in E4599 and AVAiL. Further analyses will be presented. Discussion: We report the first plasma BM data in a phase III placebo-controlled randomized study in mBC with Bv. In this subset analysis, using a novel ELISA assay, VEGF and VEGFR2 were identified as promising candidates for predictive value for PFS. A similar observation has been made in an independent BM analysis of pancreatic cancer (AVITA; data on file). Plasma data have so far not shown these trends in mCRC and mNSCLC. Further prospective evaluation of these markers is warranted. Authors D.W.M. and S.L.d.H. contributed equally. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-16-04.

PPARγ inhibits inflammatory reaction in oxidative stress induced human diploid fibloblast

The ageing of an inevitable life function is an unavoidable regressive physical process. Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear hormone receptor family. PPARgamma plays an important role in regulating several metabolic pathways. Recently, PPARgamma has been implicated in inflammatory responses and age-related diseases. The aim of this study was to determine the anti-inflammatory reaction of PPARgamma in an induced ageing progress. The late passage of human diploid fibroblasts (HDF), an in vitro ageing model, reveals the biological index materials of ageing. Aged cells showed decreased PPARgamma expression and elevated levels of intracellular adhesion molecule-1 (ICAM-1), an inflammatory molecule. To induce the aged cell phenotype, the middle stage of HDF cells (PD31) were induced stress induced premature senescence (SIPS) with 200 microM H(2)O(2) for 2 h. SIPS-HDF cells showed high levels of ICAM-1, extracellular signal regulated kinase (ERK1/2) activity and matrix metallomatrix protease (MMP-2, -9) activity, and low levels of PPARgamma expression. A reconstitution of SIPS HDF cells with Ad/PPARgamma resulted in the downregulation of ICAM-1, ERK1/2, MMP-2 and -9, and normalized growth of SIPS-HDF cells. Moreover, PPARgamma in aged HDF cells reduced pro-inflammatory molecules and eliminated the formation of reactive oxygen species (ROS) through the ERK1/2 pathway. These results strongly suggest that PPARgamma plays a key role in age-related inflammation and may have clinical applications as a molecular target in the treatment of age-related inflammation.

Rat Pancreatic β Cells and Cytomegalovirus Infection

Cytomegalovirus (CMV) infection has been suggested to accelerate beta-cell destruction and thereby to contribute to new-onset diabetes and failure of islet allografts in both humans and rodents. Surprisingly, direct CMV infection of beta cells has received only minor attention. Therefore, we investigated the susceptibility of rat beta cells for rat CMV (RCMV) infection and the direct effects on the regulation of immune cell-activating ligands. Primary rat beta cells, the rat beta-cell line Rin-m5F, and fibroblasts were RCMV-infected in vitro. The viral gene and protein expression levels were determined as a measure for RCMV susceptibility. Gene expression levels of intracellular adhesion molecule 1, lymphocyte function associated antigen 3, rat major histocompatibility complex region A, rat major histocompatibility complex region E, toll like receptor 2, and clustered domain 14 were determined as a measure for cellular immunogenicity. We demonstrate that beta cells are susceptible for RCMV infection but allow only low levels of viral gene expression. In contrast, infected fibroblasts demonstrated productive viral infection and formation of viral progeny. After RCMV infection, beta-cell immunogenicity was markedly increased, as demonstrated by the increased cellular expression of immune cell-activating ligands. Direct beta-cell infection by RCMV and subsequent low-grade viral gene expression may lead to increased immunogenicity of native or transplanted beta cells in vivo. An infection-induced enhanced beta-cell recognizability may have important consequences for beta-cell survival and the development of diabetes or rejection of islet grafts.

Macronutrient Composition and Increased Physical Activity Modulate Plasma Adipokines and Appetite Hormones during a Weight Loss Intervention

We have shown previously that in overweight premenopausal women, changes in macronutrient composition and increasing the number of steps walked per day favorably affect body composition and plasma lipid profiles. As a follow-up, we evaluated the effect of moderate carbohydrate intake and increased physical activity on inflammation and regulation of appetite. Seventy premenopausal women with a body mass index (BMI) between 25 and 37 kg/m(2) participated in a 10-week weight loss intervention program consisting of the following macronutrient energy distribution: 40% carbohydrate, 30% fat, and 30% protein, in addition to a progressive increase in the number of steps taken per day. Plasma adiponectin, intracellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), leptin, and ghrelin levels were assessed at baseline and after 10 weeks. Subjects reduced body weight by 4.5%, waist circumference (WC) by 6.4%, and trunk fat by 4.6%. Plasma insulin and insulin resistance assessed by homeostasis model assessment (HOMA) were reduced after 10 weeks (p < 0.01). Plasma adiponectin was increased by 11% (p < 0.05), and ICAM-1 levels were decreased (p < 0.05) after 10 weeks. A negative correlation was found between changes in insulin and changes in adiponectin between baseline and 10 weeks (r = -0.397, p < 0.01), indicating a role of adiponectin in increasing insulin sensitivity. In addition, plasma ghrelin levels were increased by 17% (p < 0.001), indicating a signal for increased appetite associated with weight loss. These studies indicate that weight loss interventions involving moderate changes in dietary carbohydrate and increases in physical activity favorably affect insulin sensitivity and decrease inflammation.

The adhesion molecules of L-selectin and ICAM-1 in thrombocytosis and thrombocytopenia

Thrombopoiesis is regulated by a variety of cytokines. Intracellular adhesion molecules are inducible cell-surface glycoproteins that belong to the immunoglobulin superfamily. Cytokines, endothelium and adhesive molecules represent the point of crosstalk in normal and pathological hematopoiesis. With the hypothesis that circulating intracellular adhesion molecule-1 (ICAM-1) and lymhocyte adhesion molecule-1 (L-selectin) concentrations could be changed based on pathological thrombopoiesis resulting in quantitative platelet disorders, we evaluated ICAM-1 and L-selectin levels in patients with thrombocytosis, thrombocytopenia and healthy controls. The L-selectin levels were found to be significantly higher in the thrombocytopenia group compared to the control group. ICAM-1 levels were found to be significantly higher in both thrombocytopenia and thrombocytosis groups compared to control group. Our study corroborates our original hypothesis implying the roles of adhesion molecules in the challenging status of pathological thrombopoiesis.

Upregulation of PTEN by CKD712, a synthetic tetrahydroisoquinoline alkaloid, selectively inhibits lipopolysaccharide-induced VCAM-1 but not ICAM-1 expression in human endothelial cells

Aims We examined our hypothesis that CKD712, (S)-1-(α-naphthylmethyl)-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline, selectively inhibits vascular adhesion molecule-1 (VCAM-1) but not intracellular adhesion molecule-1 (ICAM-1) expression via activation of PTEN in human umbilical vein endothelial cells (HUVECs) activated with lipopolysaccharide (LPS). Methods and results In order to do this, cells were pretreated with CKD712 1 h prior to stimulate with LPS (1 μg/ml) for 16 h, and VCAM-1 and ICAM-1 levels were measured by Western blot. We found that CKD712 dose-dependently inhibited VCAM-1 but not ICAM-1 expression after LPS stimulation in HUVECs. Furthermore CKD712 blocked the attachment of monocytes (U397) to HUVECs by LPS. Differential effect of CKD712 on adhesion molecules was mediated via regulation of PI3K/Akt signaling. It was found that CKD712 is able to significantly upregulate PTEN activity through its dephosphorylation. The inhibitory effect of CKD712 in activated HUVECs was reversed by siPTEN. In addition, administration of CKD712 (10 mg/kg) inhibited VCAM-1 but not ICAM-1 expression in thoracic aorta of LPS (10 mg/kg)-treated rats. Conclusion Our results indicate that upregulation of PTEN by CKD712 selectively inhibit VCAM-1 expression in LPS-treated HUVECs. Thus CKD712 may be beneficial in the treatment of cardiovascular disorders such as atherosclerosis.

Inhibitory effect of betulinic acid on TNF‐α‐induced adhesion molecule and its related signaling

Vascular inflammation is an important event in the development of vascular diseases including atherosclerosis. In the present study, we investigated whether betulinic acid (BA) inhibits vascular inflammation process induced by tumor necrosis factor‐alpha (TNF‐α) in human umbilical vein endothelial cells (HUVEC). Pretreatment with BA was significantly blocked TNF‐α‐induced expression levels of cell adhesion molecules such as vascular cell adhesion molecule‐1 (VCAM‐1), intracellular adhesion molecule‐1 (ICAM‐1), and endothelial cell selectin (E‐selectin). Gelatin zymography results showed that TNF‐α‐induced metalloproteinase (MMP)‐2 and MMP‐9 activities were prevented by pretreatment with BA. Real time qRT‐PCR data also showed that BA suppressed TNF‐α‐induced increase in VCAM‐1, ICAM‐1, and E‐selectin mRNA expression levels. In addition, BA inhibited the TNF‐α‐induced intracellular reactive oxygen species (ROS) production. Furthermore, pretreatment with BA significantly inhibited the translocation to nucleus and transcriptional activity of NF‐κB increased by TNF‐α. In conclusion, the present data suggested that BA could suppress TNF‐α‐induced vascular inflammatory process, due to the inhibition of ROS and NF‐κB activation in HUVEC.

Phosphomannopentaose sulfate (PI-88) inhibits retinal leukostasis in diabetic rat

Retinal leukostasis, mediated by intracellular adhesion molecule-1 (ICAM-1) and vascular endothelial growth factor (VEGF), has been implicated in the pathogenesis of early diabetic retinopathy. Phosphomannopentaose sulfate (PI-88) is a highly sulfonated oligosaccharide which inhibits heparanase activity and competes with heparan sulfate binding to growth factors. In this study, we evaluated whether PI-88 could inhibit retinal leukostasis in strepotzotocin(STZ)-induced diabetic rat and elucidated the possible mechanisms. Diabetes was induced in Sprague–Dawley rats by intraperitoneal injection (i.p.) of STZ. Three months after induction, diabetic rats were administered PI-88 (25 mg/kg body weight) or vehicle solution daily via i.p. for 14 consecutive days. Leukostasis was analyzed on retinal flatmounts by concanavalin A and CD45 immunofluorescence staining. Retinal function was analyzed by electroretinography (ERG). ICAM-1 and VEGF levels in retinas were studied by Western blot and enzyme-linked immunosorbent assay (ELISA) respectively. The systemic administration of PI-88, but not vehicle, significantly decreased the number of adherent leukocytes in retinas by 52.24% ( P < 0.001) and led to significant preservation (about 50%, P < 0.001) of scotopic ERG a- and b-wave amplitudes in treated diabetic rats as compared to those of diabetic control rats. These changes were associated with downregulation of ICAM-1 (45%, P < 0.001) and VEGF (26.83 ± 2.01 versus 40.8 ± 3.24 pg/mg, P < 0.01) in retinas of PI-88 treated diabetic rats as compared to those of diabetic control rats. PI-88 significantly inhibited retinal leukostasis and reversed retinal dysfunction by a mechanism that may include decreased ICAM-1 and VEGF expression in diabetic rats. Our data suggests that PI-88 is a promising agent for the treatment of diabetic retinopathy.

Anti-Inflammatory Effect ofGastrodia elataRhizome in Human Umbilical Vein Endothelial Cells

Vascular inflammation is a pivotal factor of a variety of diseases, such as atherosclerosis and tumor progression. The present study was designed to examine the anti-inflammatory effect of ethanol extract of Gastrodia elata rhizome (EGE) in primary cultured human umbilical vein endothelial cells (HUVEC). Pretreatment of cells with EGE attenuated TNF-alpha-induced increase in expression levels of cell adhesion molecules such as intracellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin. Real time qRT-PCR also showed that EGE decreased the mRNA expression levels of ICAM-1, VCAM-1, E-selectin as well as macrophage chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8). In addition, EGE significantly inhibited TNF-alpha-induced increase in monocyte adhesion of HUVEC in a dose-dependent manner. Furthermore, EGE significantly inhibited TNF-alpha-induced intracellular reactive oxygen species (ROS) production and p65 NF-kappaB activation by preventing IkappaB-alpha phosphorylation. In conclusion, the present data suggest that EGE could suppress TNF-alpha-induced vascular inflammatory process via inhibition of oxidative stress and NF-kappaB activation in HUVEC.

The Effects of Atorvastatin on Hematological and Inflammatory Parameters.

Background: Chronic inflammation may play role in the development of atherosclerosis and its complications. In hypercholesterolemia, the evidences of chronic inflammation such as the stimulation of chemokines and cytokines, increase in endothelial adhesion molecules, and the immune reactions against oxidants on lipoproteins are detected. It was shown that statins had some beneficial effects on lipids, thrombosis, endothelial dysfunction, and smooth muscle proliferation.Aim: To investigate the effects of atorvastatin on hematological and inflammatory parameters in prospective study.Material and methods: Forty patients (14 male and 26 female) with primary hypercholesterolemia were treated with 20 mg/day atorvastatin for 12 weeks, according to Adult Treatment Panel for Third Report of National Cholesterol Education Program. All patients received a detailed description of this study and signed consent prior to enrollment. National and local ethical committees approved this study. The exclusion criteria were secondary hypercholesterolemia, acute coronary syndromes, liver and renal dysfunctions, diabetes mellitus, acute/chronic infection and inflammatory diseases, pregnancy, lactation, malignancy, and tendency to bleeding. At baseline, and 12th weeks, lipid levels such as low-density lipoprotein cholesterol (LDL-C), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), very-low-density lipoprotein cholesterol (VLDL-C), triglycerides (TGs), and hematological parameters such as whole blood cell counts, hemoglobin and fibrinogen levels, CD3, CD4, CD5, CD8, CD14, CD16, CD19, CD40, CD45 using flow-cytometry, inflammatory parameters such as interleukin-1 (IL-1), IL-6, IL-18, tumor necrosis factor-alpha (TNF-a), interferon-gamma, soluble CD-40, intracellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1, high-sensitive CRP, sedimentation rate, and enzymes including CK, AST, ALT were evaluated. The results were compared with two-paired student's-t test.Results: At the end of study, atorvastatin decreased TC (p<0.001), LDL-C (p<0.001), TGs (p=0.006), VLDL-C (p=0.012), and HDL-C (p<0.001). While absolute lymphocyte (p=0.003) and platelet counts (p=0.001) were decreased with atorvastatin treatment, absolute monocyte count increased (p=0.002). On flow-cytometric examination, the expressions of CD14 (p=0.015) and CD19 (p=0.039) on lymphocytes were decreased with atorvastatin. Moreover, atorvastatin decreased the levels of TNF-a (p<0.001), sCD40 (p<0.001), ICAM-1 (p<0.001), and IL-18 (p=0.024). ALT increased at the end of treatment (p=0.041). But important adverse events were not seen in the patients.Conclusion: The anti-platelet and anti-inflammatory effects of atorvastatin, independent from lipid-lowering effects, may play an important role on the prevention of atherosclerosis, in addition to its beneficial effects on lipid parameters.