Endothelin-1 Staining Research Articles

Abstract 11964: The Effect of Endothelin-1 on Angiotensin II-Induced Renal Injury

Introduction: Hypertension (HT) and kidney disease is increasingly recognized as a global public health problem. Angiotensin II (AngII) induces endothelin-1 (ET-1), and ET-1 is a key mediator of kidney fibrosis. Hypothesis: Inhibition of ET-1 may contribute to modulation of HT and renal injury induced by angiotensin II. Methods: In addition to wild-type (WT) mice, we also used IL-1 receptor antagonist-deficient (IL-1Ra -/- ) mice that were prone to AngII-induced renal inflammation, because WT (C57BL) mice were genetically resistant to renal injury in response to AngII. Both mice were infused with AngII (1000ng/kg/min) using subcutaneous osmotic pumps for 14 days. Results: Systolic blood pressure in IL-1Ra -/- mice significantly increased compared with WT mice (197±5 vs 169±9 mmHg, p<0.05). Furthermore, renal preproendothelin-1 mRNA expression and serum levels of ET-1 were also significantly higher in IL-1Ra -/- mice (p<0.05). Immunostaining revealed that ET-1 was elevated in glomeruli and tubules in the cortex of IL-1Ra -/- mice compared with WT mice (Figure upper panels: ET-1 staining), indicating elevated ET-1 might contribute to glomerular injury (Figure middle panels: PAS staining) and intratubule fibrosis (Figure lower panels: Elastica Masson staining) detected in the histological analysis. Finally, we investigated whether pharmacological inhibition of ET receptors could prevent AngII-induced renal injury in IL-1Ra -/- mice. Bosentan significantly decreased AngII-induced HT, and protected renal injury in IL-1Ra -/- mice. On the other hand, treatment of hydralazine, an antihypertensive agent, did not suppress renal damage in AngII-infused IL-1Ra -/- mice in spite of improvement of HT. Conclusions: Our results indicate that inhibition of ET-1 might protect AngII-induced renal inflammation beyond an antihypertensive therapy, suggesting that we might be required to develop new strategies for blocking ET-1 to prevent hypertensive renal injury.

COMPARATIVE EFFECT OF ATENOLOL AND NEBIVOLOL ON REVERSE REMODELING OF INTRAMURAL CORONARY ARTERIOLES IN THE SPONTANEOUSLY HYPERTENSIVE RAT MODEL

Objective: Arterial hypertension is an established major risk factor for ischemic heart disease. Functional and structural abnormalities at the level of the microcirculation may play a crucial role in coronary microvascular dysfunction. Indeed, coronary flow reserve is often reduced at angiography in hypertensive patients independently on the presence of left ventricular hypertrophy (Mancini M. et al. Microcirculation 2017). The aim of this study was to assess the effect of the beta-blocker Atenolol and the selective third-generation beta1-adrenergic antagonist Nebivolol on markers of remodeling and endothelial dysfunction in coronary microvascular. Nebivolol has been demonstrated to possess direct vasodilator and antioxidant properties which may improve endothelial dysfunction and reduce smooth muscle cell proliferation. Design and method: For this purpose, we used three groups of SHR rats: Hypertensive untreated group (SHR); SHR plus Atenolol 50 mg/kg/day by oral gavaging for 8 weeks (SHR + ATE); SHR plus Nebivolol 20 mg/kg/day by oral gavaging for 8 weeks (SHR + NEB). Protein expression levels of vasoactive factors, oxidative stress and remodeling markers in coronary microcirculation were determined by immunohistochemical analysis of Endothelin-1 (ET-1), inducible nitric oxide synthase (iNOS) and transforming growth factor-beta1 (TGF-beta1). Results: No differences were observed in structural parameters of mesenteric arteries among groups. SHR + NEB showed a significant reduction in ET-1, iNOS, and TGF-beta1 staining in coronary small arteries comparing to SHR and SHR + ATE. Conclusions: In comparison to atenolol, nebivolol may provide additional benefit beyond blood pressure reduction and beta-receptor blockade since show greater activity in the modulation of protein involved in vasodilation and oxidative stress protection.

Endothelin-1 indicates unfavorable prognosis in primary high-grade non-muscle-invasive urothelial bladder cancer.

To conduct a prospective study of the potential prognostic role of endothelin-1 (ET-1) in a cohort of primary high-grade non-muscle-invasive urothelial bladder cancer patients, who were treated with adjuvant intravesical Bacillus Calmette-Guérin (BCG). Patients with primary high-grade nonmuscle- invasive urothelial bladder cancer, who received postoperatively induction and maintenance BCG therapy, were prospectively included. Recurrence and progression were histologically proven. Immunohistochemical staining for ET-1 was assessed. Epidemiological, pathological and clinical parameters as well as the expression of ET-1 in tumor specimens were statistically analyzed for recurrence, progression, recurrence-free survival (RFS) and progression-free survival (PFS). ET-1 associates significantly with recurrence (p = 0.000), progression (p = 0.000), RFS (p = 0.000) and PFS (p = 0.000). The patient's age is also significant for recurrence (p = 0.003, OR = 1.273 95% CI: 1.086-1.492) and RFS (p = 0.013). ET-1 seems to deteriorate prognosis in patients suffering from primary high-grade non-muscle-invasive urothelial bladder cancer, who are treated with adjuvant BCG instillations. Furthermore, the patient's age associates with an increased likelihood for recurrence.

Protective Effect of Ocotillol, the Derivate of Ocotillol-Type Saponins in Panax Genus, against Acetic Acid-Induced Gastric Ulcer in Rats Based on Untargeted Metabolomics.

Gastric ulcer (GU), a prevalent digestive disease, has a high incidence and is seriously harmful to human health. Finding a natural drug with a gastroprotective effect is needed. Ocotillol, the derivate of ocotillol-type saponins in the Panax genus, possesses good anti-inflammatory activity. The study aimed to investigate the gastroprotective effect of ocotillol on acetic acid-induced GU rats. The serum levels of endothelin-1 (ET-1) and nitric oxide (NO), the gastric mucosa levels of epidermal growth factor, superoxide dismutase and NO were assessed. Hematoxylin and eosin staining of gastric mucosa for pathological changes and immunohistochemical staining of ET-1, epidermal growth factor receptors and inducible nitric oxide synthase were evaluated. A UPLC-QTOF-MS-based serum metabolomics approach was applied to explore the latent mechanism. A total of 21 potential metabolites involved in 7 metabolic pathways were identified. The study helps us to understand the pathogenesis of GU and to provide a potential natural anti-ulcer agent.

Evaluation of the gastroprotective effects of 20 (S)-ginsenoside Rg3 on gastric ulcer models in mice

BackgroundGastric ulcer (GU) is a common gastrointestinal disease that can be induced by many factors. Finding an effective treatment method that contains fewer side effects is important. 20 (S)-ginsenoside Rg3 is a kind of protopanaxadiol and has shown superior antiinflammatory and antioxidant effects in many studies, especially cancer studies. In this study, we examined the treatment efficacy of 20 (S)-ginsenoside Rg3 on GU. MethodsThree kinds of GU models, including an alcohol GU model, a pylorus-ligated GU model, and an acetic acid GU model, were used. Mouse endothelin-1 (ET-1) and nitric oxide (NO) levels in blood and epidermal growth factor (EGF), superoxide dismutase, and NO levels in gastric mucosa were evaluated. Hematoxylin and eosin staining of gastric mucosa and immunohistochemical staining of ET-1, inducible nitric oxide synthase (NOS2), and epidermal growth factor receptors were studied. Ulcer index (UI) scores and UI ratios were also analyzed to demonstrate the GU conditions in different groups. Furthermore, Glide XP from Schrödinger was used for molecular docking to clarify the interactions between 20 (S)-ginsenoside Rg3 and EGF and NOS2. Results20 (S)-ginsenoside Rg3 significantly decreased the UI scores and UI ratios in all the three GU models, and it demonstrated antiulcer effects by decreasing the ET-1 and NOS2 levels and increasing the NO, superoxide dismutase, EGF, and epidermal growth factor receptor levels. In addition, high-dose 20 (S)-ginsenoside Rg3 showed satisfactory gastric mucosa protection effects. Conclusion20 (S)-ginsenoside Rg3 can inhibit the formation of GU and may be a potential therapeutic agent for GU.

Biomechanical stress in myocardial infarctions: can endothelin-1 and growth differentiation factor 15 serve as immunohistochemical markers?

Myocardial infarctions go along with biomechanical stress, i.e. stretching of muscle fibres, and the expression of certain marker molecules. We tested if two of those markers, endothelin-1 (ET-1) and growth differentiation factor 15 (GDF-15), can be used as immunohistochemical markers for myocardial ischaemia/infarctions. The study included experiments with an animal model, the isolated perfused Langendorff heart, as well as the investigation of human tissue samples drawn during autopsies. The overall picture of our results showed that GDF-15 is very sensitive and expressed very fast, not only as a consequence of ischaemia/infarctions, but also under other circumstances. Even an expression only caused by agony had to be discussed. ET-1, on the other hand, was less sensitive but only positive in those human cases with ischaemia/infarction that also showed typical alterations in conventional histology. Therefore, both markers did not proof to be a suitable diagnostic tool for myocardial infarctions. However, positive staining for ET-1 was also seen in rats' hearts that suffered from arrhythmias after electric shock and in the myocardium of the right ventricle in human control cases in which a right heart failure has to be discussed. Thus, especially ET-1 should be subject of further studies that focus on these pathologies.

Endothelin-1 expression in prostate needle biopsy specimens correlated with aggressiveness of prostatic cancer

As the prostate adenocarcinoma is one of the most common malignant tumors in males, looking for a marker to effectively predict aggressiveness and metastatic potential in an apparently localized cancer in initial needle biopsy specimens can help the clinicians to make more appropriate decision for treatment, planning, and choosing appropriate targeted therapy. The present study assessed the value of Endothelin-1 expression to predict prognosis of prostatic cancer. In a cross sectional study, 83 patients who underwent radical prostatectomy in Hasheminejad Kidney Center in 2008 through 2012 were assigned to two groups including 43 with and 40 without extra-prostatic extension (EPE). Endothelin-1 staining was performed on Paraffin Embedded blocks of preoperative needle biopsies. The expression of Endothelin-1 increased in 72% of patients in the group with EPE (P<0.001). The group with Endothelin-1 positivity showed higher serum level of prostate specific antigen (PSA) (p = 0.039). Endothelin-1 expression was positive in 67% of patients with perineurial invasion (P<0.001). Adjusting the baseline variables of PSA and PN in a multivariable logistic regression model, the Endothelin-1 positivity could effectively predict EPE in patients with prostatic cancer (OR: 5.46, p = 0.010). Correlation of Endothelin-1 expression in needle biopsy specimens in expected with extra-prostatic extension of tumor in radical prostatectomy specimens, perineurial invasion and serum PSA level at the time of diagnosis.

Molecular analysis of vascular smooth muscle cells from patients with giant cell arteritis: Targeting endothelin-1 receptor to control proliferation.

The pathophysiology of giant cell arteritis (GCA) and the mechanisms underlying vascular remodeling, are poorly understood. We aimed to compare vascular smooth muscle cells (VSMCs) from patients with GCA and controls by a proteomic and gene expression profile approach and to identify the signaling pathways involved in proliferation. VSMCs were cultured from temporal artery biopsies (TABs) from patients with biopsy-proven GCA (TAB+-GCA), biopsy-negative GCA (TAB--GCA), and diagnosis other than GCA (GCA-control). VSMCs from normal human aorta (HAoSMC) were used as controls. 2D-differential in-gel electrophoresis and Affymetrix chips were used to compare proteomes and gene expression profiles of VSMCs. Proliferation was assessed by BrdU incorporation assay. TAB+-GCA and GCA-control TABs underwent immunohistochemistry staining for endothelin-1 (ET-1) and receptors ETAR and ETBR. We identified 16, 30 and 2 protein spots differentially expressed between TAB+-GCA and GCA-control VSMCs, TAB+-GCA and TAB--GCA VSMCs and TAB--GCA and GCA-control VSMCs, respectively (fold change ≥1.5 and p≤0.05). Among the 153 proteins differentially expressed between TAB+-GCA and HAoSMC VSMCs, many were linked with ET-1. Genes differentially expressed between TAB+-GCA and GCA-control VSMCs were involved in proliferation. ET-1 was identified as a link between genes of interest. Proliferation was reduced for TAB+-GCA VSMCs on treatment with the endothelin antagonist macitentan and its active metabolite. Patients showing transmural expression of ET-1 in temporal artery lesions received a significantly higher glucocorticoid daily dose after 6-month follow-up. Inhibiting the proliferation with macitentan, combined with glucocorticoids, might be a promising therapeutic approach for patients with GCA.

Effect of acupuncture on the genetic expression of myocardial endothelin-1 and atrial natriuretic peptide in rats with stress-induced prehypertension

ObjectiveTo explore the protective effect of acupuncture against myocardial injury in rats with stress-induced prehypertension (SIPH) by observing the genetic expression of myocardial endothelin-1 (ET-1) and atrial natriuretic peptide (ANP). MethodsThirty-six Wistar rats were randomized into three groups: the control group, model group, and model + acupuncture (AP) group (n = 12 rats per group). During the 11-day modeling period, the model group and the model + AP group experienced plantar electric stimulation in combination with noise exposure, and daily acupuncture intervention was applied simultaneously in the model + AP group; the control group did not experience modeling or acupuncture. Systolic pressure (SP) was measured the day before modeling began, and on the 3rd, 5th, 7th, 9th, and 11th day after modeling initiation. On the 12th day, histopathological observation of the left ventricle was made with hematoxylin-eosin staining and quantitative genetic expression of myocardial ET-1, and ANP was tested by quantitative real-time polymerase chain reaction (PCR). ResultsSP was higher in the model group than the control group on the 3rd, 5th, 7th, 9th, and 11th days (all P < .01). SP in the model + AP group was lower than that in the control group on the 5th and 7th days (respectively, P = .008, P = .002) and on the 9th and 11th days (P = .029, P = .039). Hematoxylin-eosin staining showed normal myocardial cellular structure in the control group. The model group showed disordered arrangement of cardiac cells with morphological changes in some nuclei, including enlargement or dissolution; there was also infiltration of inflammatory cells and proliferation of collagen fibers. In the model + AP group, most of the myocardial cells were normally structured, and only part of the cells had morphological changes with enlarged nuclei or dissolution. Real-time PCR showed that expression of ET-1 and ANP mRNA in the model group was greater than the control group (respectively, P = .024, P = .000101). The model + AP group had lower expression of ET-1 and ANP mRNA compared with the model group (respectively, P = .033, P = .043). ConclusionAcupuncture may lower blood pressure and downregulate the genetic expression of myocardial ET-1 and ANP in SIPH rats, suggesting a protective effect of acupuncture against myocardial damage.

Differential changes in vascular mRNA levels between rat iliac and renal arteries produced by cessation of voluntary running

Early vascular changes at the molecular level caused by adoption of a sedentary lifestyle are incompletely characterized. Herein, we employed the rodent wheel-lock model to identify mRNAs in the arterial wall that are responsive to the acute transition from higher to lower levels of daily physical activity. Specifically, we evaluated whether short-term cessation of voluntary wheel running alters vascular mRNA levels in rat conduit arteries previously reported to have marked increases (i.e. iliac artery) versus marked decreases (i.e. renal artery) in blood flow during running. We used young female Wistar rats with free access to voluntary running wheels. Following 23 days of voluntary running (average distance of ∼15 km per night; ∼4.4 h per night), rats in one group were rapidly transitioned to a sedentary state by locking the wheels for 7 days (n = 9; wheel-lock 7 day rats) or remained active in a second group for an additional 7 days (n = 9; wheel-lock 0 day rats). Real-time PCR was conducted on total RNA isolated from iliac and renal arteries to evaluate expression of 25 pro-atherogenic and anti-atherogenic genes. Compared with the iliac arteries of wheel-lock 0 day rats, iliac arteries of wheel-lock 7 day rats exhibited increased expression of TNFR1 (+19%), ET1 (+59%) and LOX-1 (+31%; all P < 0.05). Moreover, compared with renal arteries of wheel-lock 0 day rats, renal arteries of wheel-lock 7 day rats exhibited decreased expression of ETb (-23%), p47phox (-32%) and p67phox (-19%; all P < 0.05). These data demonstrate that cessation of voluntary wheel running for 7 days produces modest, but differential changes in mRNA levels between the iliac and renal arteries of healthy rats. This heterogeneous influence of short-term physical inactivity could be attributed to the distinct alteration in haemodynamic forces between arteries.

Endothelin-1 and Endothelin A Receptor Immunoreactivity Is Increased in Patients with Diabetic Nephropathy

Background: Endothelin-1 (ET-1) is associated with progression of renal disease, acting as a vasoconstrictor and growth factor for mesangial cells. ET-1 and endothelin A receptor (ET-RA) might have a role in the development of diabetic nephropathy (DN). The aims of this study were to determine ET-1 and ET-RA expressions in patients with DN and to correlate these expressions with renal function and proteinuria. Materials and methods: This is a cross-sectional study comprising 13 patients with type 2 diabetes mellitus and DN, 10 patients with proteinuric IgA nephropathy, and 13 samples of normal kidney from tumor nephrectomies. Demographic and selected data were collected from medical charts. The distribution and intensity of ET-1 and ET-RA immunostaining in renal biopsies were determined by immunohistochemistry and these correlated with the estimated glomerular filtration rate (eGFR) and proteinuria. Results: Patients with DN and IgA nephropathy on biopsy had markedly increased staining for ET-1 in endothelial cells of glomerular and peritubular capillaries when compared with controls (p < 0.001). ET-RA staining was also more intense and more diffuse in DN and IgA nephropathy than in controls (p = 0.019) and was restricted to tubular epithelial cells. A positive correlation was observed between ET-1 expression and proteinuria (r = 0.634, p = 0.027), but both ET-1 and ET-RA expressions did not correlate with eGFR. Conclusion: In this preliminary report, the higher expressions of ET-1 and ET-RA found in both DN and IgA nephropathy suggest a potential role for the endothelin system in DN as well as in other nondiabetic glomerular diseases.

P5-01-19: Endothelin-1 Expression in Breast Cancer Tissue, Surrounding Stroma, Correlation with Tumor Microvessel Density and Clinical Outcome.

Abstract Background: Endothelin-1 (ET-1) is a peptide which regulates normal biological processes such as vascular tone. In addition, endothelin signaling pathway is dysregulated in pathophysiological conditions such as cancer and fibrosis. It has been shown that endothelin-1 is expressed in breast cancer tissue but little is known with regard to ET-1 expression in surrounding tumor stroma. In the present study, we investigated ET-1 expression in breast tumor cells, surrounding stroma, association with tumor microvessel density (MVD) and impact on clinical outcome. Materials and Methods: We conducted a retrospective, multicenter study. Patients from 3 medical centers with histologically documented stage I-III invasive breast cancer were included in the study. Paraffin embedded formalin fixed breast cancer tissue and surrounding stroma were evaluated for ET-1 and CD34 (marker for MVD) by IHC. ET-1 cytoplasmic expression was scored as positive (3+ by IHC) or negative (0, 1+, 2+). Stained vessels by CD34 were counted in five consecutive fields at 40 x magnification and their mean was recorded. Demographics, clinical data and recurrence free interval (RFI) in months were available for statistical analysis. Results: The study included 92 patients with average age of 55 years at diagnosis. Median follow up of patients at the time of analysis was 72 months. Total of 29 patients experienced disease progression ( 17 locoregional and 12 distant). Tumor ET-1 expression positively correlated with earlier stage: odds ratio (OR) = 22 for stage I and OR=20 for stage II as compared with stage III. Positive ET-1 staining in tumor was detected in 72.8 % of cases, while ET-1 positive expression in stroma was detected only in 6.5% of cases. Interestingly, all ET-1 stroma positive tumors were estrogen receptor (ER) positive. The means of CD34 were not different according to ET-1 expression either in the tumor or stroma. Triple negative breast cancer tumors exhibited higher MVD (p=0.0177). In the logistic regression model relating ET-1 expression in tumor to clinical variables, ET-1 positive tumors showed a trend for the association with higher relapse rate (p=0.058). Multivariate analysis suggested that there was no significant difference in the recurrence free interval (RFI) between the ET-1 positive and ET-1 negative groups (long-rank test p-value = 0.71). Survival analysis identified stage as a significant predictor of RFI in the Cox proportional hazard model that included ET-1 expression and other clinical variables. Conclusions: The significant predictor for ET-1 expression in the tumor was early stage. High tumor ET-1 expression was more common in patients who experienced breast cancer recurrence. No association was found between ET-1 expression and MVD, and between ET-1 expression and time to recurrence. Further studies with larger sample size are needed to better delineate a role of ET-1 as prognostic biomarker in early stage breast cancer. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-01-19.

Changes of Endothelin-1 in Rats Corpus Cavernosum Smooth Muscle of Partial Bladder Outlet Obstruction

Purpose: Lower urinary tract symptoms and erectile dysfunction (ED) are common disorders in men and increase with age. Recently, there has been evidence to suggest that patients with high benign prostatic hyperplasia symptom scores have an increased frequency of ED. This study was performed to evaluate the change of the expression of endothelin-1 (ET-1) in the corpus cavernosum by inducing partial bladder outlet obstruction (PBOO) in a rat model. Materials and Methods: Eight Sprague-Dawley male rats were used for induction of PBOO by placing a 25 gauge angioneedle sheath around the urethra, then ligating the bladder neck with 3-0 vicryl suture and six sham-operated rats were ligated very loosely without causing obstruction. At three and five weeks after PBOO, the distal portion of the ligated penile tissues was dissected for immunohistochemical staining of ET-1 in the vascular endothelial cell. Results: ET-1 was weakly expressed (+) in the smooth muscle layers of the control group and a very strong expressed (++++) in the experimental group on 3 weeks after PBOO. However, there was no expression in the experimental group on 5 weeks after PBOO. There was no expression (?) in the vascular endothelial cells of the control group and very strong expression (++++) in the experimental group on 3 weeks after PBOO. Howerver, there was no expression in the experimental group on 5 weeks after PBOO. Conclusions: The changes of penis by inducing partial bladder outlet obstruction in the rats were associated with increased ET-1 in the vascular and peri-vascular smooth muscle cells of the corpus cavernosum.

Endothelin‐1: a predictor of extracapsular extension in clinically localized prostate cancer?

To assess the value of endothelin-1 (ET-1) expression in predicting extracapsular extension (ECE) in clinically localized prostate cancer (PCa). ET-1 expression was determined by immunohistochemistry on archival needle biopsies (NBs) from 94 patients (49 pT2 and 45 pT3a) who underwent radical prostatectomy (RP) for clinical T1-T2 PCa. Each sample was analysed independently by two pathologists blinded to the clinical data. In univariate analysis, high ET-1 expression in NBs, pre-operative prostate-specific antigen (PSA) level >10 ng/ml, percentage of positive biopsy cores and NB Gleason score ≥7 were significantly associated with ECE as determined on subsequent RP. No significant association was found between clinical stage and ECE. In multivariate analysis, there was a significant association with high ET-1 expression in NBs (p = 0.006), pre-operative PSA level >10 ng/ml (p = 0.049), and NB Gleason score ≥7 (p = 0.002). These three pre-operative factors combined provided the best model for predicting ECE with 93.3% sensitivity, 49% specificity, 62.5% positive predictive value, 88.9% negative predictive value. The combination yielded a higher concordance index (0.760 vs 0.720) and offered a higher log partial likelihood than the same model without ET1 (112.8 vs 105.7, p = 0.01). ET-1 expression was strongly associated with ECE and, when combined with pre-operative PSA level and Gleason score, improved the predictive accuracy of pre-operative NBs. Its assessment in patients with localized PCa might be useful when making treatment decisions. Further studies with standardisation of immunohistochemical staining and multi-institutional validation are now needed to establish the appropriate use of ET-1 staining in PCa staging and to evaluate inter-observer reproducibility.

Endothelin-1 expression in scleroderma renal crisis

The objective of the study was to investigate the role of endothelin-1 in the pathogenesis of scleroderma renal crisis in patients with systemic sclerosis. We used immunohistochemical analysis with anti-endothelin-1 and anti-von Willebrand factor antibodies in comparing kidney biopsies from patients with systemic sclerosis and scleroderma renal crisis (n = 14); from normal kidneys (n = 5); and from patients with typical hemolytic uremic syndrome and thrombotic microangiopathy (n = 5), antiphospholipid syndrome (n = 6), diabetic nephropathy (n = 5), minimal change disease with cyclosporine toxicity (n = 5), or nephroangiosclerosis (n = 5). Kidney biopsies from all systemic sclerosis patients presented specific lesions: glomerular lesions with thickened capillary walls (n = 6, 42.8%), mesangiolysis (n = 3, 21.4%), fibrin thrombi (n = 3, 21.4%), hypertrophy of juxtaglomerular apparatus (n = 5, 35.7%), arteriolar lesions showing mucinous intimal thickening and lumen mucoid occlusions (n = 13, 92.8%), proliferation of intimal cells (ie, "onion-skin" lesions; n = 13, 92.8%), fibrinoid necrosis (n = 3, 21.4%), and fibrin thrombosis (n = 4, 28.6%). Chronic lesions in large arteries showed modifications such as fibrous intimal thickening (n = 13, 92.8%). The pattern of endothelial staining for endothelin-1 in both glomeruli and arteriolar lesions appears to be specific for scleroderma renal crisis. Glomerular endothelin-1 staining without arteriolar staining was seen in hemolytic uremic syndrome; and isolated arteriolar staining (without glomerular staining) was seen in a number of conditions including antiphospholipid nephropathy, cyclosporine toxicity, and diabetic nephropathy. Endothelin-1 is overexpressed in glomeruli and arterioles of patients with scleroderma renal crisis, which suggests that endothelin-1 might be a therapeutic target in this condition.

552 IMMUNOHISTOCHEMICAL EXPRESSION AND PROGNOSTIC VALUE OF ENDOTHELIN-1 AND ENDOTHELIN-A RECEPTOR IN CLINICALLY LOCALIZED PROSTATE CANCER

You have accessJournal of UrologyProstate Cancer: Basic Research IV1 Apr 2010552 IMMUNOHISTOCHEMICAL EXPRESSION AND PROGNOSTIC VALUE OF ENDOTHELIN-1 AND ENDOTHELIN-A RECEPTOR IN CLINICALLY LOCALIZED PROSTATE CANCER Marie-Dominique Azémar, Johann Ménard, Anne Durlach, Renaud Fay, Philippe Birembaut, and Frédéric Staerman Marie-Dominique AzémarMarie-Dominique Azémar Reims, France More articles by this author , Johann MénardJohann Ménard Reims, France More articles by this author , Anne DurlachAnne Durlach Reims, France More articles by this author , Renaud FayRenaud Fay Nancy, France More articles by this author , Philippe BirembautPhilippe Birembaut Reims, France More articles by this author , and Frédéric StaermanFrédéric Staerman Reims, France More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2010.02.772AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES We examined the expression of endothelin-1 (ET-1) and endothelin-A receptor (ET-A-R) in clinically localized prostate cancer and we investigated the utility of ET-1 and ET-A-R immunostaining in the prediction of biochemical failure after radical prostatectomy (RP). METHODS This retrospective study was carried out in 90 patients who had undergone RP for clinically localized prostate cancer (pT2 and pT3a, with negative surgical margins). Biochemical relapse was defined as serum PSA value ≤ 0.2 ng/ml, concerning 27 patients of the 90 included. Mean follow-up was 62.9±26.2 months. A semi-quantitative immunohistochemical study of ET-1 and ET-A-R was performed on RP specimens by one pathologist blinded to the clinical data. Adenocarcinoma tissue and normal gland adjacent to tumor regions were both analyzed for each specimen. For ET-1, intensity of immunoreactivity was scored from 0 (no staining) to 3 (strong staining). For ET-A-R, the stronger intensity of the core was scored from 0 to 3 and percentage of cells stained with this intensity were reported. Then a combined score for ET-A-R was determined by multiplying the intensity and percentage (from 0 to 300). RESULTS Mean age at radical prostatectomy was 64.7±6.5 years. Mean preoperative serum PSA was 9.0±5.3 ng/ml. Extraprostatic extension (pT3a) was found in 16.7% of RP specimens. Gleason sum was < 7 in 53.9% of the specimens. Median time to biochemical failure was 35.8 months. ET-1 staining intensity was moderate to strong in 80% of carcinoma tissue and in only 2.2% of normal adjacent to tumor regions. ET-A-R stronger intensity was moderate to strong in 90.8% of carcinoma tissue and in 65.5% of normal adjacent to tumors regions. Mean preoperative serum PSA (p=0.0008), Gleason sum (p=0.005), and ET-A-R expression in tumoral tissue (p=0.0012) were significantly higher in case of recurrence. In univariate analysis, preoperative PSA level (p=0.008), extracapsular extension (p=0.013) and ET-A-R immunostaining combined score in tumoral tissue (p=0.006) were statistically significant. ET-1 expression was not a prognostic marker of biochemical relapse. In multivariate analysis, only Gleason sum (p=0.013) and ET-A-R overexpression in tumoral glands (p=0.026) were independent prognostic factors of biochemical failure after RP. CONCLUSIONS ET-1 and ET-A-R are overexpressed in clinically localized prostate cancer. In this study, ET-A-R expression on RP specimen is an independent marker of biochemical failure after RP. © 2010 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 183Issue 4SApril 2010Page: e217 Advertisement Copyright & Permissions© 2010 by American Urological Association Education and Research, Inc.MetricsAuthor Information Marie-Dominique Azémar Reims, France More articles by this author Johann Ménard Reims, France More articles by this author Anne Durlach Reims, France More articles by this author Renaud Fay Nancy, France More articles by this author Philippe Birembaut Reims, France More articles by this author Frédéric Staerman Reims, France More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Expression immunohistochimique de l’endothéline-1 sur biopsies prostatiques : marqueur pronostique d’adénocarcinome localement avancé ?

The study evaluated the immunohistochemistry expression of endothelin-1 (ET-1) by prostate cancer (PCa) in prostate biopsies as an extracapsular stage (pT3a) prognostic factor. Sixty-eight radical prostatectomies (RP) were performed for clinically localised PCa (35 pT2 and 33 pT3a according to the 2002 pTNM classification). Age, digital rectal examination, initial PSA, biopsy Gleason score, positive biopsies ratio, specimen Gleason score, biopsy and RP specimen perineural neoplasic invasion, PCa DNA ploidy, PCa Ki-67 DNA image cytometry and biopsy and RP specimen ET-1 immunohistochemistry expression for both group were compared. Semi-quantitative ET-1 staining assessment was realised by the same pathologist. pT3a group initial PSA was higher (p=0.032). No statistically difference was noticed between pT2 and pT3a groups for positive biopsies ratio, biopsy perineural neoplastic invasion and biopsy DNA ploidy determination. Biopsy Gleason score > or =7 was predictive of a pT3a stage (p=0.03). Statistically higher intensity of ET-1 PCa expression was observed in biopsies and specimens in pT3a group than in pT2 group (p<0.001 and p=0.01). In multivariate analysis, biopsy ET-1 PCa expression was an independent risk factor of pT3a stage with specificity 79 %, sensibility 69 %, predictive positive value 77 % and negative positive value 72 %. Combined with initial PSA > or =7, values were respectively 100 %, 76.9 %, 100 % and 57.1 %. Endothelin-1 (ET-1) prostate cancer biopsy expression in our study was an independent prognostic factor of extracapsular stage (pT3a). Further studies will assess the relevance of ET-1 expression study in clinically localised PCa for active surveillance, curative treatment or targeted adjuvant therapy management.

Maternal EDN1 G5665T polymorphism influences circulating endothelin-1 levels and plays a role in determination of preeclampsia phenotype

Preeclampsia, a pregnancy-specific syndrome characterized by hypertension and proteinuria, has a familial tendency and is an important cause of maternal and neonatal mortality. Abnormal endothelin-1 synthesis and/or release can explain abnormalities seen in preeclampsia. We prospectively evaluated whether endothelin-1 levels are increased in preeclampsia, to verify if placenta is the source, and any association between the maternal EDN1 G5665T single-nucleotide polymorphism and circulating endothelin-1 levels and preeclampsia manifestation. A total of 120 with preeclampsia and 118 normotensive primigravid patients with singleton pregnancy were enrolled. Preeclampsia was defined as new onset of elevated blood pressure greater than 140/90 mmHg and at least 2+ proteinuria on two occasions at least 4 h apart after 20 weeks of gestation in previously normotensive pregnant women. Patients were excluded if they had hypertension before 20 weeks of gestation, diabetes, asthma, heart disease, kidney disease, hematological disorder, autoimmune disease, urinary tract infection, current or past history of smoking, twin pregnancy, molar pregnancy or eclampsia. Genotyping was done using a PCR-RFLP-based method. Placenta samples were collected from 20 preeclampsia and 24 normotensive pregnancies. Reverse transcriptase PCR was done for preproendothelin and beta-actin mRNA. Placental endothelin-1 staining was assessed by immunohistochemistry in villous and extravillous trophoblasts and endothelium. Plasma endothelin-1 levels were measured using ELISA. The preeclampsia group showed higher circulating endothelin-1 levels (1.45 +/- 0.55 vs. 0.91 +/- 0.42 pg/ml; P < 0.0001), reduced frequency of GG genotype (34 vs. 49%; P = 0.025) and increased T allele frequency (0.43 vs. 0.28; P = 0.04). A significant association was noted between endothelin-1 levels and blood pressure in the entire cohort and in the group with preeclampsia (P < 0.001). Circulating endothelin-1 levels were higher in those bearing even one copy of the T variant (1.08 +/- 0.48 vs. 1.31 +/- 0.59 pg/ml; P = 0.004). Placental endothelin-1/beta-actin mRNA ratio was significantly reduced (0.91 +/- 0.77 vs. 3.20 +/- 1.68; P < 0.001) and endothelin-1 staining was lower (P < 0.001) in placental endothelium in preeclampsia. Maternal endothelin-1 is elevated and correlates with the severity of blood pressure elevation in preeclampsia. The endothelin-1 is likely released from the maternal endothelium. Presence of T allele at 5665 position in maternal EDN1 gene is associated with higher endothelin-1 levels. Placental endothelin-1 synthesis is reduced in preeclampsia. The combination of elevated maternal and reduced placental endothelin-1 may be an adaptive response to reduced uteroplacental flow in preeclampsia.

Immunolocalisation of endothelin-1 in human brain

The potent vasoconstrictor endothelin-1 (ET-1) may function as a neuropeptide and be a contributing factor in some neurological disorders, e.g. Alzheimer’s dementia. The presence of ET-1 has been studied more extensively in the rat and porcine nervous systems than in the human brain. Also, the recent description of the extensive ET-1 mRNA localisation in human neural tissue supports expression in regions of human brain not previously investigated. Using specific anti-ET-1 polyclonal antiserum, we immunolocalised ET-1 in 24 regions of human brain autopsy tissues, and correlated this with ET-1 mRNA distribution. ET-1 immunoreactivity was observed within some cells of all the 24 areas examined. Neuronal staining for ET-1 was demonstrated within the diencephalon, brainstem, basal nuclei, cerebral cortex, cerebellar hemisphere, amygdala and hippocampus. In addition, ET-1 immunolabelling was visualised in the pituitary gland as well as in the choroid plexus. The primary sensory cortex and pineal gland also contained immunoreactive ET-1, although ET-1 mRNA had never been detected in these regions previously. The localisation of ET-1 and its subsequent correlation with ET-1 mRNA in most of the regions investigated suggest a more extensive distribution of the ET system in the human brain than was previously identified.

VASCULAR REMODELLING IN THE INTERNAL MAMMARY ARTERY GRAFT AND ASSOCIATION WITH ELEVATED ENDOTHELIN-1 EXPRESSION

Introduction: The vasoconstricting peptide Endothelin-1 (ET-1) has been associated with atherosclerotic cardiovascular disease, AAA, hypertension and hypercholesterolemia. It is known to stimulate quiescent vascular smooth muscle cells (VSMC) into the growth cycle and has been linked to intimal thickening following endothelial injury and is associated with vessel wall remodelling in salt-sensitive hypertension models. Enhanced ET-1 expression has been reported in the internal mammary artery (IMA) and was markedly higher in patients undergoing cardiac bypass surgery who were diabetic and /or hypercholesterolemic. Aims: To firstly review the histopathology of the IMA and secondly, determine the relationship between ET-1 expression in this vessel and mitogenic activity in the medial VSMC. Methods: Vessel tissue collected at the time of CABG surgery was formalin-fixed and paraffin-embedded for histological investigation. Cross sections of the left distal IMAwere stained with Alcian Blue/Verhoeff’s van Gieson to assess medial degeneration and identify the elastic lamellae and picrosirius red to determine the collagen content (specifically type I and type III). Immunohistochemistry staining was used to assess VSMC growth (PCNA label), tissue ET-1 expression, VSMC (SMCa-actin) area and macrophage/monocyte (anti-CD68) infiltration. Quantitative analysis was performed to measure the VSMC area in relation to ET-1 staining. Results: Fifty-five IMA specimens from the CABG patients (10F; 45M; mean age 65 years) were collected for this study. Fourteen donor IMAspecimens were used as controls (7F; 7M; mean age 45 years). Significant medial hypertrophy, VSMC disorganisation and elastic lamellae destruction was detected in the CABG IMA. The amount of Alcian blue staining in the CABG IMA was almost double that of the control (31.85+/14.52% Vs 17.10+/9.96%, P= .0006). Total collagen and type I collagen content was significantly increased compared with controls (65.8+/18.3% Vs 33.7 + / 13.7%, P= .07), (14.2 + /10.0% Vs 4.8 + /2.8%, P= .01), respectively. Tissue ET-1 and PCNA labelling were also significantly elevated the CABG IMA specimens relative to the controls (69.99 + /18.74%Vs 23.33 + /20.53%, P= .0001, and 37.29 + /12.88% Vs 11.06 + /8.18, P= .0001), respectively. There was mild presence of macrophages and monocytes in both CABG and control tissue. Conclusions: The IMA from CABG patients has elevated levels of type I collagen in the extracellular matrix indicative of fibrosis and was coupled with deleterious structural remodelling. Abnormally high levels of ET-1 were measured in the medial SMC layer and was associated with VSMC growth but not related to any chronic inflammatory response within the vessel wall.