Marker Of Endothelial Cell Injury Research Articles

Thrombomodulin as a promising Marker of Endothelial Dysfunction in Hemodialysis Patients with Non Alcoholic Fatty Liver Disease

Abstract Background Non-alcoholic fatty liver disease (NAFLD) is associated with endothelial dysfunction which is a common problem in haemodialysis patients and risk for high cardiovascular mortality in haemodialysis. Thrombomodulin is a promising marker of endothelial cell injury in different pathological conditions. This study evaluates the additive effect of non-alcoholic fatty liver disease (NAFLD) in prevalent HD (hemodialysis) patients on endothelial dysfunction. Methods This is a case-control study which was conducted on 60 ESRD (end-stage renal disease) patients on conventional hemodialysis and 30 apparently healthy controls. Patients were divided into 2 groups: group A (N = 30): with NAFLD& group B (N = 30): without NAFLD, excluding elderly, diabetic, chronic liver disease, advanced heart failure, active infection, COVID-19 infection and autoimmune disease& patients withhemodialysiscatheters. Thrombomodulin was measured for all participants by ELISA technique. Results Thrombomodulin had the ability to detect endothelial dysfunction in patients with NAFLD at Cut off value <0.8 ng/ml with sensitivity 53.33, specificity 80, PPV 72.7 and NPV 63.2. On comparing Thrombomodulin in patients with NAFLD(4.378±3.762 ng/ml), Non NAFLD (1.126±0.591 ng/ml)and control (0.755±0.314ng/ml) there was significant difference (P- value<0.001). Post hoc analysis showed significantly high Thrombomodulin level in patients with NAFLD compared to Non NAFLD and control with p-value<0.001 while there was no significant difference between patients with (Non NAFLD and control) p-value 0.792. NAFLD patients with higher Thrombomodulin level have 2.5 times higher risk to have endothelial dysfunction. There was no significant correlation between Thrombomodulin and all variables in the study. Conclusions Thrombomodulin can be used as specific marker for endothelial dysfunction in hemodialysis patients with Non Alcoholic fatty liver.

Dihydroartemisinin ameliorates cerebral I/R injury in rats via regulating VWF and autophagy-mediated SIRT1/FOXO1 pathway.

Dihydroartemisinin (DHA) has been found to inhibit the expression of von Willebrand factor (VWF), a marker of endothelial cell injury, but its mechanism in cerebral ischemia/reperfusion (I/R) injury remains obscure. In this study, I/R model was constructed through middle cerebral artery occlusion (MCAO) in rats, followed by DHA administration. The effect of DHA on rat cerebral I/R injury was investigated by 2,3,5-triphenyltetrazolium chloride staining, hematoxylin and eosin staining, TUNEL staining, and Western blot. Brain microvascular endothelial cells (BMVECs) isolated from newborn rats were exposed to oxygen-glucose deprivation/reoxygenation (OGD/R), and then treated with DHA. The results showed that MCAO treatment induced infarction, nerve cell apoptosis, and brain tissue impairment in rats, which was mitigated by DHA. OGD/R inhibited viability and accelerated apoptosis of BMVECs, which was alleviated by DHA. I/R procedures or OGD/R up-regulated expressions of VWF, ATG7, Beclin1, and LC3-II/LC3-I ratio, while down-regulating Occludin, Claudin-5, ZO-1, P62, SIRT1, and FOXO1 expressions in vivo and in vitro; however, these effects of I/R procedures or OGD/R were offset by DHA. VWF overexpression reversed the above effects of DHA on OGD/R-induced BMVECs. In summary, DHA ameliorates cerebral I/R injury in rats by reducing VWF level and activating autophagy-mediated SIRT1/FOXO1 signaling pathway.

#2779 THROMBOMODULIN IS A SPECIFIC MARKER OF ENDOTHELIAL DYSFUNCTION IN HEMODIALYSIS PATIENTS WITH NON-ALCOHOLIC FATTY LIVER DISEASE

Abstract Background and Aims Non-alcoholic fatty liver disease (NAFLD) is associated with endothelial dysfunction which is a common problem in hemodialysis patients and a risk for high cardiovascular mortality in hemodialysis. Thrombomodulin is a promising marker of endothelial cell injury in different pathological conditions. This study evaluates the additive effect of non-alcoholic fatty liver disease (NAFLD) in prevalent HD patients on endothelial dysfunction. Methods A case-control study was conducted on 60 end-stage renal disease (ESRD) patients on conventional hemodialysis: group A: 30 patients with NAFLD, group B: 30 patients without NAFLD, and control group of 30 apparently healthy subjects. Excluding elderly, diabetic patients, chronic Liver disease, advanced heart Failure, active infection, COVID-19 infection, autoimmune diseases and patients with hemodialysis catheters. Thrombomodulin was measured for all participants by ELISA technique. Results Thrombomodulin could detect endothelial dysfunction in patients with NAFLD at Cut off value <0.8 ng/ml with sensitivity 53.33%, specificity 80%, PPV 72.7%, and NPV 63.2%. On comparing Thrombomodulin in patients with NAFLD (4.378±3.762 ng/ml), Non-NAFLD (1.126±0.591 ng/ml), and control (0.755±0.314 ng/ml) there was a significant difference (P-value<0.001). Post hoc analysis showed significantly high Thrombomodulin levels in patients with {NAFLD versus Non-NAFLD} and {NAFLD versus control} with p-value<0.001, 0.001 respectively while there was no significant difference between patients with Non-NAFLD and control p-value 0.792. NAFLD patients with positive thrombomodulin have a risk of 2.5 times having endothelial dysfunction. There was no significant correlation between Thrombomodulin and all variables in the study. Conclusions Thrombomodulin can be used as specific marker for endothelial dysfunction in hemodialysis patients with Non Alcoholic fatty liver disease.

HYDROCORTISONE, ASCORBIC ACID, AND THIAMINE THERAPY DECREASE RENAL OXIDATIVE STRESS AND ACUTE KIDNEY INJURY IN MURINE SEPSIS.

Background: Acute kidney injury (AKI) occurs frequently in septic patients and correlates with increased mortality. Because clinical studies investigating hydrocortisone, ascorbic acid, and thiamine (HAT) have demonstrated discordant results, studies were performed using mortality stratification for therapy to identify candidates for therapy and determine mechanisms of organ injury. Methods: Sepsis was induced using the cecal ligation and puncture (CLP) model of sepsis with fluid and antibiotic support. Heart rate (HR) measurements obtained 6 hours after CLP stratified mice into live predicted (P-Live) or die predicted (P-Die). Stratified mice were then randomized for treatment with HAT or vehicle given 7 hours after CLP. Physiologic measurements were taken again at 24 hours, and mice were killed to collect blood and organs. Results: The following five groups were created: (1) P-Live vehicle, (2) P-Live HAT, (3) P-Die vehicle, (4) P-Die HAT, and (5) naive mice. Comparisons were made to test the hypotheses that (1) P-Die vehicle mice will have significant deterioration compared with P-Live mice targeting the kidney and (2) HAT will correct these deleterious changes in P-Die mice. Compared with P-Live, P-Die mice had a significant decline in all measured physiologic parameters (HR, cardiac output, breath rate, and temperature), which were corrected with HAT therapy (P < 0.05 for all parameters). The P-Die mice had declines in the ascorbic acid within the blood, peritoneal lavage, and kidney homogenate compared with P-Live mice indicating consumption, and the decline was corrected with HAT. Elevated IL-6, KC, Macrophage Inflammatory Protein-2, and IL-1RA were found in P-Die mice and decreased with HAT. Markers of endothelial cell injury (glypican 1 and glypican 4) were elevated in the P-Die mice, and these values were decreased with HAT therapy. Low oxygen levels with subsequent oxidative stress (OS) in the kidney were visualized in histologic sections using hypoxyprobe and also with carbonyl proteins and 8-iso-prostaglandin F2α in kidney homogenates. The P-Die mice had significant elevations of renal OSs, which was ameliorated with HAT. Kidney injury was evident in the P-Die mice compared with P-Live mice with elevations in blood urea nitrogen and cystatin C, which were significantly reduced with HAT. There was no evidence of global hypoxia or organ injury because hepatic parameters remained normal. Conclusions: Our data show that in CLP-induced sepsis, P-Die mice have increased inflammation, OS, and kidney injury. Hydrocortisone, ascorbic acid, and thiamine therapy decreased renal OS and injury in the P-Die group when given after the onset of sepsis-induced physiologic changes.

Prognostic Significance of Plasma Hepatocyte Growth Factor in Sepsis.

To assess any correlation of plasma hepatocyte growth factor (HGF) levels with relevant endothelial cell injury parameters and determine the prognostic value in septic patients. A prospective, observational study was conducted in patients with sepsis admitted to the Department of Critical Care Medicine at the Zhongda Hospital from November 2017 to March 2018. Plasma HGF levels were measured by enzyme-linked immunosorbent assay in the first 24 h after admission (day 1) and on day 3. The primary endpoint was defined as all-cause 28-day mortality. Furthermore, we analyzed the correlation of HGF with relevant endothelial cell injury markers. Eighty-six patients admitted with sepsis were included. HGF levels of nonsurvivors were elevated compared to those of survivors on day 1 (1940.62 ± 74.66 pg/mL vs. 1635.61 ± 47.49 pg/mL; P = 0.002) and day 3 (1824.82 ± 137.52 pg/mL vs. 1309.77 ± 83.49 pg/mL; P = 0.001) and showed a strong correlation with von Willebrand factor (r = 0.45, P < 0.0001), lactate (r = 0.35, P = 0.0011), pulmonary vascular permeability index (r = 0.38, P = 0.0241), first 24 h fluid administration (r = 0.38, P < 0.0001), and sequential organ failure assessment score (r = 0.40, P = 0.0001). Plasma HGF levels were able to prognostically discriminate between survivors and nonsurvivors on day 1 (AUC: 0.72, 95%CI: 0.60-0.84) and day 3 (AUC: 0.77, 95%CI: 0.63-0.91). HGF levels are associated with sepsis and correlated with established markers of endothelial cell injury. Elevated HGF levels in sepsis patients are an efficient indicator of poor prognosis. The study was registered in Clinical Trial (Registration Number: NCT02883231).

Effect of heparin on histone-mediated the expression of von Willebrand factor and fibrinogen in lung tissue

To observe the effects of histones on lung injury and von Willebrand factor (vWF) and fibrinogen (FIB) levels in mice, and to explore the protective effect of heparin. Twenty-four male C57BL/6 mice aged 6-10 weeks were divided into control group, histone group and histone+heparin group according to random number table method with 8 mice in each group. The mice in the histone group were injected with histone (50 mg/kg) via the tail vein, and the mice in the histone+heparin group were injected with unfractionated heparin (400 U/kg) via the tail vein at 1 hour after administration of histone, and those in the control group were given the same amount of normal saline. Four hours after histone injection, the lungs of the mice were harvested and the lung wet/dry weight ratio (W/D) and the pulmonary water contents were measured. The pathological changes in lung tissue were observed by hematoxylin and eosin (HE) staining under microscope, and the extent of lung injury was evaluated. The positive expression of vWF which was the marker of endothelial cell injury was observed by immunohistochemistry. The real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to determine the expression level of FIB mRNA in lung tissue. The lung W/D ratio and pulmonary water contents in the histone group were significantly higher than those in the control group [lung W/D ratio: 6.19±0.53 vs. 4.54±0.25, pulmonary water contents: (82.59±2.03)% vs. (78.52±1.51)%, both P < 0.01]. The lung W/D ratio and pulmonary water contents in the histone+heparin group were significantly lower than those in the histone group [lung W/D ratio: 4.84±0.35 vs. 6.19±0.53, pulmonary water contents: (79.21±1.48)% vs. (82.59±2.03)%, both P < 0.01], indicating that the heparin could reduce histone-induced pulmonary edema. Histological examination showed that the alveolar structure of the control group was intact, and the alveolar cavity was clean without exudation. In the histone group, the lungs were significantly damaged. The alveolar wall was thickened, infiltrated by inflammatory cells and focally alveolar hemorrhage, edema, associated with alveolar fibrin deposition and micro-thrombus formation. The lung histopathological score in the histone group was significantly higher than that in the control group (5.15±0.87 vs. 0.18±0.17, P < 0.01). All of the pathological changes were significantly alleviated in the histone+heparin group, and the histopathological score of the lung was significantly lower than that in the histone group (2.28±0.72 vs. 5.15±0.87, P < 0.01), indicating that the histone-induced lung injury was improved by heparin. Immunohistochemistry showed that high vWF expressions of lung tissue were observed in the histone group while there was almost no positive expression in the control group, and the vWF expression in the histone+heparin group was significantly reduced, indicating that heparin protected mice against histone-induced endothelial cell injury. The FIB mRNA expression of lung tissue in the histone group was about 49.82 times of the control group (2-ΔΔCT: 55.30±18.84 vs. 1.11±0.45, P < 0.01), and the expression of FIB mRNA in the histone+heparin group was decreased, which was 23.87 times of the control group (2-ΔΔCT: 26.50±9.97 vs. 1.11±0.45, P < 0.01), but it was significantly lower than that in the histone group (2-ΔΔCT: 26.50±9.97 vs. 55.30±18.84, P < 0.01), indicating that heparin could inhibit histone-induced hypercoagulable environment in lung. Histone causes pulmonary edema, endothelial cell injury and coagulation activation. Heparin could effectively attenuate histone-induced lung injury and coagulation activation.

Single Cell RNA Sequencing Identifies HSPG2 and APLNR as Markers of Endothelial Cell Injury in Systemic Sclerosis Skin.

Objective: The mechanisms that lead to endothelial cell (EC) injury and propagate the vasculopathy in Systemic Sclerosis (SSc) are not well understood. Using single cell RNA sequencing (scRNA-seq), our goal was to identify EC markers and signature pathways associated with vascular injury in SSc skin.Methods: We implemented single cell sorting and subsequent RNA sequencing of cells isolated from SSc and healthy control skin. We used t-distributed stochastic neighbor embedding (t-SNE) to identify the various cell types. We performed pathway analysis using Gene Set Enrichment Analysis (GSEA) and Ingenuity Pathway Analysis (IPA). Finally, we independently verified distinct markers using immunohistochemistry on skin biopsies and qPCR in primary ECs from SSc and healthy skin.Results: By combining the t-SNE analysis with the expression of known EC markers, we positively identified ECs among the sorted cells. Subsequently, we examined the differential expression profile between the ECs from healthy and SSc skin. Using GSEA and IPA analysis, we demonstrated that the SSc endothelial cell expression profile is enriched in processes associated with extracellular matrix generation, negative regulation of angiogenesis and epithelial-to-mesenchymal transition. Two of the top differentially expressed genes, HSPG2 and APLNR, were independently verified using immunohistochemistry staining and real-time qPCR analysis.Conclusion: ScRNA-seq, differential gene expression and pathway analysis revealed that ECs from SSc patients show a discrete pattern of gene expression associated with vascular injury and activation, extracellular matrix generation and negative regulation of angiogenesis. HSPG2 and APLNR were identified as two of the top markers of EC injury in SSc.

Expression of Endothelial Cell Injury Marker Cd146 Correlates with Disease Severity and Predicts the Renal Outcomes in Patients with Diabetic Nephropathy

Background/Aims: Glomerular endothelial cell injury plays a crucial role in the development of diabetic nephropathy (DN). CD146, an endothelial marker, was shown to increase in chronic kidney disease (CKD), but its role in DN remains unknown. We aim to assess whether CD146 could be used to evaluate disease severity and predict renal outcomes in DN at early stages. Methods: 159 non-dialysis type 2-DN patients from 2008 to 2015 were enrolled to measure the plasma concentration of soluble CD146 (sCD146). 94 type 2 diabetes mellitus patients without DN and 100 healthy subjects were used as controls. The patients with CKD stage 1-3 were referred as early stage patients. Another independent cohort of 48 patients with biopsy-proved DN was used for the immunohistochemistry study of CD146. Renal outcomes were defined as doubling of serum creatinine, initiation of renal replacement therapy or death. Results: We found that plasma level of sCD146 was upregulated and associated with renal function in DN patients. sCD146 was proved to be a more optimal marker than urine albumin creatinine ratio to evaluate disease severity in these DN patients. The kidney expression of CD146 was co-localized with endothelial marker CD31 and increased in DN. CD146 staining in kidney was correlated with the severity of pathological changes in DN patients. Survival analysis suggested that both plasma and biopsy expression of CD146 were correlated with renal outcomes. Conclusions: CD146 is associated with kidney injury and could be a good marker to predict renal outcomes in patients with early stages of DN.

Endothelial progenitor cells in chronic obstructive pulmonary disease and emphysema.

Endothelial injury is implicated in the pathogenesis of COPD and emphysema; however the role of endothelial progenitor cells (EPCs), a marker of endothelial cell repair, and circulating endothelial cells (CECs), a marker of endothelial cell injury, in COPD and its subphenotypes is unresolved. We hypothesized that endothelial progenitor cell populations would be decreased in COPD and emphysema and that circulating endothelial cells would be increased. Associations with other subphenotypes were examined. The Multi-Ethnic Study of Atherosclerosis COPD Study recruited smokers with COPD and controls age 50–79 years without clinical cardiovascular disease. Endothelial progenitor cell populations (CD34+KDR+ and CD34+KDR+CD133+ cells) and circulating endothelial cells (CD45dimCD31+CD146+CD133-) were measured by flow cytometry. COPD was defined by standard spirometric criteria. Emphysema was assessed qualitatively and quantitatively on CT. Full pulmonary function testing and expiratory CTs were measured in a subset. Among 257 participants, both endothelial progenitor cell populations, and particularly CD34+KDR+ endothelial progenitor cells, were reduced in COPD. The CD34+KDR+CD133+ endothelial progenitor cells were associated inversely with emphysema extent. Both endothelial progenitor cell populations were associated inversely with extent of panlobular emphysema and positively with diffusing capacity. Circulating endothelial cells were not significantly altered in COPD but were inversely associated with pulmonary microvascular blood flow on MRI. There was no consistent association of endothelial progenitor cells or circulating endothelial cells with measures of gas trapping. These data provide evidence that endothelial repair is impaired in COPD and suggest that this pathological process is specific to emphysema.

Abstract 18190: Associations Between Endothelial Injury, Severity of Post-cardiac Arrest Syndrome and Mortality After Out-of-hospital Cardiac Arrest

Background: Post-cardiac arrest syndrome (PCAS) is characterized by systemic inflammation and endothelial dysfunction, which may exacerbate organ failure and may be related to adverse outcome. We sought to investigate the associations between trombomodulin (TM), a marker of endothelial cell injury, and severity of PCAS and the possible prognostic importance in comatose survivors of out-of-hospital cardiac arrest treated with targeted temperature management (TTM). Methods: We included 163 patients, randomly assigned to TTM at 33°C or 36°C for 24 h in the TTM-trial at a single center. Serum TM was analyzed at 0, 24, 48 and 72 hours after cardiac arrest and stratified in median value of TM for outcome analysis at each time point to a high or low TM group. Severity of PCAS was assessed daily by the Sequential Organ Failure Assessment score. Survival status was recorded at 180 days. Results: Corresponding values of TM and severity of PCAS correlated at each time point, baseline (r=0.31, p=0.0001), 24 h (r=0.52, p&lt;0.0001), 48 h (r=0.50, p&lt;0.0001) and 72 h (r=0.36, p&lt;0.0001). Levels of TM changed significantly within the first 3 days (p&lt;0.0001) with no significant difference between temperature groups, p interaction =0.32. At 24 h, the 180-day mortality rate in patients with high TM levels (&gt; 6.9 ng·ml -1 = median) was 42% compared to 18% in patients with low TM levels, p=0.002 (Figure), corresponding to an unadjusted hazard ratio of 2.6 (95%CI: 1.4-4.9), p=0.003. When adjusting for pre-hospital prognostic factors the hazard ratio for high TM levels at 24 h was 1.5 (95%CI: 0.7-3.1), p=0.31, with no significant interaction between level of TM and TTM-group, p=0.74. Analysis of TM at 48 and 72 hours showed similar results. Conclusions: Levels of TM are significantly elevated and associated with severity of PCAS with no effect of target temperature at 33°C or 36°C. High TM levels were associated with increased mortality, but the association was explained by confounding factors.

Label-Free and Sensitive Detection of Thrombomodulin, a Marker of Endothelial Cell Injury, Using Quartz Crystal Microbalance.

Thrombomodulin (TM), an integral glycoprotein on the surface of endothelial cells, can be released during endothelial cell injury and the levels of serum TM are regarded as an important parameter of activity in vasculitides in vivo. Quantitative detection of TM and investigation on the release of soluble thrombomodulin (sTM) by the injured HUVEC-C cells using quartz crystal microbalance (QCM) were achieved in this work. Anti-antibody (AAb) and bovine serum albumin (BSA) were bound on gold nanoparticles (GNPs) to construct BSA-GNPs-AAb nanocomposites and they were characterized by transmission electron microscope, UV-vis, and infrared spectrophotometry, respectively. The capture of the nanocomposites on the TM antibody modified electrode, which was tested by scanning electron microscope, could result in a great decrease of the resonant frequency (f0). This binding was effectively inhibited by the beforehand immobilized TM proteins on the electrode surface due to the strong steric hindrance effect. It led to the decrease of the frequency changing extent. The relative frequency-shift was found to be proportional to the logarithm of the TM concentration from 10 to 5000 ng mL(-1) with a detection limit of 2 ng mL(-1). By analyzing the growth medium used for cell incubation, the release of sTM by the injured HUVEC-C cells in the presence of H2O2 was confirmed. The sTM amount in the growth medium was increased with the enhancement of contact time of the cells with H2O2, proving that sTM may serve as a specific marker of endothelial cell injury.

YKL-40, a Marker of Cardiovascular Disease and Endothelial Dysfunction, in Kidney Transplant Recipients

BackgroundYKL-40 is an inflammatory glycoprotein involved in endothelial dysfunction and expressed in macrophages in the earliest lesion of atherosclerosis. Because cardiovascular mortality is the main cause of death, there are no data on kidney transplant recipients, so the aim of the study was to assess YKL-40 in that population, with particular attention being paid to the relationship with endothelial damage. MethodsWe studied 68 patients after kidney transplantation. Complete blood count, creatinine, lipids, and fasting glucose were studied with the use of standard laboratory methods. We assessed YKL-40; markers of inflammation high-sensitivity C-reactive protein (hs-CRP), von Willebrand factor, thrombomodulin, interleukin-6, intracellular adhesion molecule, and vascular cell adhesion molecule; markers of hemostasis plasmin-antiplasmin complexes, thrombin-antithrombin complexes, prothrombin fragments 1+2; and marker of kidney function neutrophil gelatinase–associated lipocalin (NGAL) with the use of commercially available assays. ResultsMean levels of YKL-40 were significantly higher in kidney allograft recipients than in the control group (P < .001). YKL-40 was also significantly higher in patients with coronary artery disease, hypertension, and diabetes compared with their counterparts without these diseases. YKL-40, in univariate analysis, was related to NGAL, protein Z, plasmin-antiplasmin complex, mean corpuscular volume, cyclosporine level, and hs-CRP. YKL-40 was not related to serum lipids, markers of endothelial cell injury, or causes of end-stage renal failure. YKL-40 was predicted by cyclosporine level, NGAL, and hs-CRP, explaining 37% of the variation. ConclusionsElevated YKL-40 may contribute to enhanced risk of cardiovascular complication, mainly owing to endothelial cell injury and inflammation in patients after kidney transplantation treated with calcineurin inhibitors.

Nitrosonifedipine Ameliorates the Progression of Type 2 Diabetic Nephropathy by Exerting Antioxidative Effects

Diabetic nephropathy (DN) is the major cause of end-stage renal failure. Oxidative stress is implicated in the pathogenesis of DN. Nitrosonifedipine (NO-NIF) is a weak calcium channel blocker that is converted from nifedipine under light exposure. Recently, we reported that NO-NIF has potential as a novel antioxidant with radical scavenging abilities and has the capacity to treat vascular dysfunction by exerting an endothelial protective effect. In the present study, we extended these findings by evaluating the efficacy of NO-NIF against DN and by clarifying the mechanisms of its antioxidative effect. In a model of type 2 DN (established in KKAy mice), NO-NIF administration reduced albuminuria and proteinuria as well as glomerular expansion without affecting glucose metabolism or systolic blood pressure. NO-NIF also suppressed renal and systemic oxidative stress and decreased the expression of intercellular adhesion molecule (ICAM)-1, a marker of endothelial cell injury, in the glomeruli of the KKAy mice. Similarly, NO-NIF reduced albuminuria, oxidative stress, and ICAM-1 expression in endothelial nitric oxide synthase (eNOS) knockout mice. Moreover, NO-NIF suppressed urinary angiotensinogen (AGT) excretion and intrarenal AGT protein expression in proximal tubular cells in the KKAy mice. On the other hand, hyperglycemia-induced mitochondrial superoxide production was not attenuated by NO-NIF in cultured endothelial cells. These findings suggest that NO-NIF prevents the progression of type 2 DN associated with endothelial dysfunction through selective antioxidative effects.

Ratio of Angiopoietin-2 to Angiopoietin-1 predicts mortality in acute lung injury induced by paraquat

BackgroundTo determine whether initial reactive oxygen species (ROS)-induced endothelial cell injury is involved in early death after paraquat intoxication and concentrations of angiopoietin-1 (Ang-1), angiopoietin-2 (Ang-2), and von Willebrand factor (VWF) reflecting endothelial cell injury, we investigated the initial endothelial cell injury marker involved in the pathogenesis of death within 5 days after paraquat ingestion.Material/MethodsSixty patients with paraquat poisoning were prospectively enrolled. Plasma samples were collected at admission. Plasma concentrations of Ang-1, Ang-2, and VWF were measured by enzyme-linked immunosorbent assay. The patients were classified into 3 categories: survivors, early death (died within 5 days after ingestion), and late death (died more than 5 days after ingestion).ResultsThe baseline concentration of Ang-2 and the Ang-2: Ang-1 ratio were significantly higher in patients who died (Ang-2 [pg/mL], 1012.75±468.02 vs. 1986.07±1675.37 [p=0.002]; Ang-2: Ang-1, 0.90±0.49 vs. 2.16±2.28 [p=0.002]). The Ang-2: Ang-1 ratio was significantly higher in the early death group (2.41±2.54) than in the survivors (0.90±0.49) and the late death group (1.33±0.64). The Ang-2: Ang-1 ratio was significantly associated with early death (OR, 2.602; 95% CI, 1.106–6.117; p=0.028) after adjusting for plasma levels of paraquat, age, PCO2, and creatinine. VWF did not predict mortality.ConclusionsEndothelial cell damage could be involved in the pathogenesis of early death following paraquat ingestion.

654: Endocan (ESM-1): a novel soluble endothelial cell injury marker in preeclampsia (PE) and intrauterine growth restriction (IUGR)

654: Endocan (ESM-1): a novel soluble endothelial cell injury marker in preeclampsia (PE) and intrauterine growth restriction (IUGR)

A potent oral P‐selectin blocking agent improves microcirculatory blood flow and a marker of endothelial cell injury in patients with sickle cell disease

Abnormal blood flow accounts for most of the clinical morbidity of sickle cell disease (SCD) [1,2]. Most notably, occlusion of flow in the microvasculature causes the acute pain crises [3] that are the commonest cause for patients with SCD to seek medical attention [4] and major determinants of their quality of life [5]. Based on evidence that endothelial P-selectin is central to the abnormal blood flow in SCD we provide results from four of our studies that are germane to microvascular blood flow in SCD. A proof-of-principle study established that doses of heparin lower than what are used for anticoagulation but sufficient to block P-selectin improved microvascular blood flow inpatients with SCD. An in vitro study showed that Pentosan Polysulfate Sodium (PPS) had greater P-selectin blocking activity than heparin. A Phase I clinical study demonstrated that a single oral dose of PPS increased microvascular blood flow in patients with SCD. A Phase II clinical study that was not completed documented that daily oral doses of PPS administered for 8 weeks lowered plasma levels of sVCAM-1 and tended to improve microvascular blood flow in patients with SCD. These data support the concept that P-selectin on the microvascular endothelium is critical to both acute vascular occlusion and chronically impaired microvascular blood flow in SCD. They also demonstrate that oral PPS is beneficial to microvascular sickle cell blood flow and has potential as an efficacious agent for long-term prophylactic therapy of SCD.

Immunohistochemical expression of von Willebrand factor in the preeclamptic placenta

Preeclampsia is a high-prevalence systemic pregnancy disorder associated with maternal and foetal mortality. Its pathogenesis is unknown, but it is thought that oxidative stress and endothelial dysfunction may play a fundamental role. Von Willebrand factor (vWF), a marker of endothelial cell injury, can be found in different cells and zones of the placenta. To determine the differential immunoexpression of vWF at different tissue types of preeclamptic placenta and endothelial dysfunction markers at maternal serum of preeclamptic pregnancies. A case-control study was performed on a population of pregnant women with preeclampsia (n = 14), and normal pregnancies (n = 8). Placental and blood plasma samples were withdrawn at delivery. Immunohistochemical vWF expression in the placental tissue was determined. Endothelial dysfunction was assessed through plasminogen activator inhibitor (PAI) 1 and 2 ratio and vWF concentration in maternal plasma. P values less than 0.05 were considered statistically significant. Preeclamptic women showed increased plasma PAI-1/PAI-2 ratio (P < 0.05). There was diminished placental vWF expression in syncytiotrophoblast and increased in the intervillous space of preeclamptic placentas (P < 0.05). No significant differences in vWF expression were found in the villous endothelium and stroma, but it was significantly higher in maternal plasma (P < 0.05). In preeclampsia occurs endothelial damage and placental cell injury. Cell damage in syncytiotrophoblast that occurs in preeclampsia could liberate vWF from syncytiotrophoblast to the placental intervillous space, and this may have pathogenic implications.

Biochemical study on some adipocyto-kines in chronic renal failure: Their relationship to endothelial dysfunction and inflammation

Chronic renal failure has been associated with impaired immunityand subclinical inflammation involving cytokines derived from adiposetissue – adipocytokines. Deteriorating renal function may increase overallinflammatory responses because of the decreased renal clearance of factors that are directly or indirectly involved in inflammation. Declining renal function may also affect the levels of additional inflammatory molecules such as C-reactive protein (CRP) and interleukin-6.The aim of the study was to assess visfatin and apelin in correlation with markers of endothelial cell injury and inflammation in 20 patients with CRF and 20 age- and sexmatched healthy controls.We assessed visfatin and apelin, markers of: coagulation: TAT (thrombin-antithrombin complexes); fibrinolysis: tPA (tissue plasminogen activator) and PAI-1 (plasminogen activator inhibitor- 1); endothelial function/injury: ICAM (intracellular adhesion molecule), VCAM (vascular cell adhesion molecule), CD40L and E-selectin and inflammation: hsCRP andIL-1â. Visfatin, apelin, TAT, ICAM, VCAM, CD40L, PAI-1, E-selectin, hsCRP, IL-1â and triglycerides were elevated while serum albumin and t-PA were decreased in CRF patients when compared with the control group.Significant positive correlations werefound between Visfatin on one hand and each of apelin, t-PA, PAI-1, Eselectin, ICAM, VCAM, hsCRP, IL-1â, CD40L and triglycerides on theother hand in patients with CRF.Also, Significant positive correlations werefound between Apelin and each of IL-1â, E-selectin, ICAM, VCAM,creatinine and triglycerides in CRF.KEY WORDS: CRF, adipocytokines, endothelial dysfunction andinflammation.

Plasma markers of oxidative stress, inflammation and endothelial cell injury in diabetic patients with overt nephropathy administered alpha-lipoic acid and angiotensin II receptor blocker

Alpha-lipoic acid (ALA) is a natural antioxidant that scavenges reactive oxygen species (ROS) and regenerates endogenous antioxidants. In the present study, plasma molecular markers of oxidative stress, inflammatory state, and endothelial cell injury was measured to investigate the effect of simultaneous blocking of both angiotensin (AT) II-and oxidative stress-mediated pathophysiologic pathways during the course of type II diabetic nephropathy in the clinical settings. Sixty-three diabetic subjects have carried out either of 12-month treatment course of AT-II receptor blocker (ARB) alone, ALA alone, or combination of both. Plasma malondialdehyde (MDA) level, a marker for oxidative stress, increased in ARB alone group whereas reduced to significantly lower level in ALA+ ARB combination group. Plasma hs-CRP level, an inflammatory marker, decreased significantly in ARB +ALA group. Plasma thrombomodulin (TM) level, a marker of endothelial cell injury, increased in both ARB and ALA group. Twenty-four hour urinary protein excretion, as a marker of renal injury, decreased in ARB and ARB+ALA group. Finally, renal function was reserved well in ARB+ALA group in compared to others. The combined administration of ALA and ARB additively attenuated or reduced plasma markers of oxidative stress, inflammation and endothelial cell injury for one year. It suggests that the benefits in attenuating atherogenic process and vascular injury retard the progression of renal dysfunction.

Persistent plasminogen activator inhibitor 1 gene expression in cardiac transplant recipients with idiopathic dilated cardiomyopathy

Plasminogen activator inhibitor 1 is the primary regulator of urokinase plasminogen activator and tissue plasminogen activator. Plasminogen activator inhibitor 1 is essential in the control of the thrombotic/fibrinolytic balance and is a marker of endothelial cell injury. Idiopathic dilated cardiomyopathy is reportedly associated with endothelial cell dysfunction. Whether endothelial cell damage plays a role in patients with dilated cardiomyopathy after cardiac transplantation remains unknown. In this study explanted hearts of cardiac transplant recipients with ischemic cardiomyopathy and dilated cardiomyopathy, as well as control myocardial tissue, were investigated for expression of urokinase plasminogen activator, tissue plasminogen activator, urokinase plasminogen activator receptor, and plasminogen activator inhibitor 1 and 2. Furthermore, plasminogen activator inhibitor 1 expression was examined in endomyocardial biopsy specimens and sera of patients with ischemic cardiomyopathy and those with dilated cardiomyopathy during the first posttransplantation year. The effect of the patient's serum on endothelial cells was assessed in vitro to examine the role of circulating endothelial cell damage-related factors. Plasminogen activator inhibitor 1 expression was upregulated in ischemic cardiomyopathy and dilated cardiomyopathy myocardial tissue versus that seen in control tissue. After transplantation, plasminogen activator inhibitor 1 expression returned to control levels in patients with ischemic cardiomyopathy. In patients with dilated cardiomyopathy, plasminogen activator inhibitor 1 expression increased at 24 weeks after transplantation in both biopsy specimens and sera versus that seen in control tissue. Sera of patients with dilated cardiomyopathy, but not that of patients with ischemic cardiomyopathy, inhibited vascular endothelial growth factor A-induced proliferation of endothelial cells, although downstream target gene activation of early growth response factor 1 and NGFI-A binding protein 2 was not affected. These data suggest for the first time that the endothelial cell damage-related process recurs in patients with dilated cardiomyopathy after transplantation, which, independently of vascular endothelial growth factor, is associated with increased plasminogen activator inhibitor 1 expression, and that this pathology might play a role in allograft remodeling in patients with dilated cardiomyopathy.