Eplerenone enhances cardioprotective effects of standard heart failure therapy through matricellular proteins in hypertensive heart failure.

AIMS: The addition of an aldosterone receptor antagonist on top of current optimal therapy (based on angiotensin II inhibition) has demonstrated an important clinical benefit in heart failure patients with systolic dysfunction. Whether this finding also applies to heart failure patients with preserved systolic function is unknown. Therefore, we have studied the effect of adding eplerenone to standard pharmacological heart failure therapy (angiotensin-converting enzyme inhibitor/angiotensin receptor blocker and diuretic and ß-blocker) in the progression of heart failure in spontaneously hypertensive heart failure (SHHF) rats. METHODS AND RESULTS: Two-month-old SHHF rats were randomized to receive no treatment (SHHF group), a standard heart failure therapy (quinapril-torasemide-carvedilol; ST-SHHF group), or the combination of eplerenone and standard heart failure therapy (Eple+ST-SHHF group) for 20 months. Untreated SHHF was characterized by progressive left ventricular hypertrophy, fibrosis, and myocardial contractile and relaxation abnormalities, leading to pulmonary congestion. Despite similar blood pressure control, the addition of eplerenone to standard heart failure therapy further prevented left ventricular hypertrophy, contractile and relaxation alterations, and pulmonary congestion than standard heart failure therapy alone. ST-SHHF and Eple + ST-SHHF rats showed similar inhibition of structural extracellular matrix proteins collagen I, collagen III and fibronectin and metalloproteinase (MMP)-2, MMP-7, MMP-12, and MMP-13. However, only the coadministration of eplerenone with standard heart failure therapy normalized the expression of matricellular proteins thrombospondin 1, tenascin C, periostin, and secreted protein acidic rich in cysteine/osteonectin to values comparable to normotensive rats. CONCLUSION: In a hypertensive heart failure rat model, the addition of eplerenone to conventional heart failure therapy further improves cardiac structural and functional parameters, delaying the progression of heart failure. These beneficial effects of eplerenone were associated with normalization of matricellular protein expression.

Ezetimibe inhibits PMA-induced monocyte/macrophage differentiation by altering microRNA expression: a novel anti-atherosclerotic mechanism.

Ezetimibe, a selective inhibitor of intestinal cholesterol absorption, effectively reduces plasma cholesterol both in monotherapy or combined with a statin. However, its effect on atherosclerosis plaque progression is certainly unknown. MicroRNAs are short non-encoding RNA molecules dynamically implicated in monocytic differentiation which is considered an essential process during atherosclerosis development. The purpose of this study was to investigate the effect of ezetimibe on monocyte/macrophage differentiation as well as the implication of microRNAs (miRNAs) in this process. THP-1 differentiation with PMA became cells adherent to the plastic surface, and induced the expression of macrophage surface markers (CD11a, CD11b and ICAM-1) and miR-155, miR-222, miR-424 and miR-503. In the presence of ezetimibe, the adhesive capacity of THP-1 cells was decreased in a dose-dependent manner (P<0.05) and the expression of CD11a, CD11b and ICAM-1 was almost totally inhibited (P<0.05). The expression of miR-155, miR-222, miR-424 and miR-503 was reduced by 55%, 100%, 75% and 100%, respectively (P<0.05). Further mechanistic studies demonstrated that ezetimibe suppressed the PMA-induced phosphorylation of ERK/MAPK and inhibited the NF-κB activity, which are upstream signalling molecules in the differentiation process. In conclusion, ezetimibe inhibits PMA-induced THP-1 cell differentiation into macrophage-like cells in association with the inhibition of miRNA pathways. Our study suggests that inhibition of miRNAs might form a novel mechanism of anti-atherosclerotic effect of ezetimibe.

Association of HIV-Infection and antiretroviral therapy with levels of endothelial progenitor cells and subclinical atherosclerosis.

BACKGROUND: Although in the general population circulating vascular progenitor cell levels have been implicated in the homeostasis of the vascular wall through differentiation into endothelium and/or smooth muscle cells, it has not yet been assessed in HIV-infected patients. We herein investigated the number of progenitor cell subpopulations in HIV-infected patients and its relationship to carotid intima-media thickness (c-IMT). METHODS: Blood samples were collected from 200 HIV-infected patients and CD34/KDR, CD34/VE-cadherin, and CD14/Endoglin progenitor cells were identified by flow cytometry. c-IMT was determined by ultrasonography. A group of 27 healthy subjects was used as control group. RESULTS: In our population (20 ART-naive patients and 180 treated patients), traditional cardiovascular risk factors were not found predictive of vascular progenitor cell levels. However, antiretroviral therapy (ART)-treatment was identified as the main predictive value for low CD34/KDR cells and high CD14/Endoglin cells after adjustment by cardiovascular risk factors (age, sex, hypertension, diabetes, and hyperlipidaemia) and HIV-related characteristics (HIV duration and ART treatment). Low levels of circulating CD34/KDR or CD34/VE-cadherin endothelial progenitor cells tended to be associated with increased c-IMT. However, a positive association was found between CD14/Endoglin cells and c-IMT. Low number of CD34/KDR cells was also associated with the longest exposure to nucleoside reverse transcriptase inhibitors and/or protease inhibitors. CONCLUSIONS: ART exposure is the main predictor of circulating vascular progenitor cell levels. However, their levels are only partially associated with high c-IMT in HIV-infected patients. ART has already been found to have proatherogenic effect, but our data first describe its relationship with vascular progenitor cells and c-IMT.

Ezetimibe inhibe la adhesión y la migración de monocitos a través de la ruta de las proteincinasas activadas extracelularmente (p44/p42 ERK1/2 / Ezetimibe suppresses adhesion and chemotaxis of human monocytes through the p44/p42ERK1/2-dependent pathway

Introducción Recientemente nuestro grupo ha demostrado que ezetimibe, un inhibidor específico de la absorción intestinal, es capaz de inhibir la inflamación vascular en un modelo de arteriosclerosis en conejo. Nuestro objetivo ha sido investigar el efecto de ezetimibe sobre la adhesión y la migración de monocitos humanos THP-1, así como la participación de la vía de señalización de las proteincinasas activadas extracelularmente (p44/p42ERK1/2) sobre el efecto observado. Material y métodos La adhesión se valoró como la capacidad de las células THP-1 para unirse a placas de cultivo, y la migración se determinó con el empleo de cámaras de quimiotaxis. La expresión de moléculas de adhesión se cuantificó mediante citometría de flujo, y la activación de p44/p42ERK1/2 se estudió mediante Western Blot. Resultados La adhesión y la migración de los monocitos THP-1 inducidas con PMA o MCP-1, respectivamente, se inhibió de forma dependiente de la dosis al preincubar las células con ezetimibe. Además, el tratamiento con ezetimibe inhibió la expresión de las integrinas CD11a y CD11b, así como la fosforilación de p44/p42ERK1/2 (la forma activa) inducida por MCP-1. Más del 90% de las células (evaluadas mediante azul tripán) eran viables tras 1 ó 2 días de exposición a ezetimibe. Conclusiones Nuestros resultados indican que ezetimibe, además de su actividad hipolipemiante, puede inhibir el proceso de adhesión y migración de los monocitos. Parece que el bloqueo de la ruta de señalización de las MAPK p44/p42ERK1/2 podría estar implicado en el efecto observado (AU)

Introduction: Recently, our group has demonstrated that ezetimibe, a specific inhibitor of intestinal absorption, is able to inhibit vascular inflammation in a rabbit model of atherosclerosis. In this study, we investigated the effect of ezetimibe on the adhesion and migration of human THP-1 monocytes in vitro. We also studied the involvement of the MAP kinase signaling pathway,p44/p42ERK1/2, as a potential mechanism responsible for the observed effect. Material and methods: Adhesion of THP-1 monocytes was measured as the ability of cells tobind to plates. Migration was studied using two-compartment chambers. The expression of adhesion molecules was assessed by flow cytometry. Activation of p44/p42ERK1/2 was measured by Western Blot. Results: Preincubation of THP-1 monocytes with ezetimibe prevented PMA-induced adhesion and MCP-1-induced migration in a dose-dependent manner. Preincubation of THP-1 monocytes with ezetimibe also inhibited the expression of the integrins CD11a and CD11b, as well asphosphorylation of p-p44/p42ERK1/2 (the active form) induced by MCP-1. More than 90% of cells(evaluated through trypan blue) were viable 1 or 2 days after exposure to ezetimibe. Conclusions: Our results indicate that, in addition to its lipid lowering activity, ezetimibe isable to inhibit the process of adhesion and migration of monocytes in vitro. Blocking of thep44/p42ERK1/2 MAPK signalling pathway seems to play a role in this anti-inflammatory effect (AU)

Rosuvastatin added to standard heart failure therapy improves cardiac remodelling in heart failure rats with preserved ejection fraction.

AIMS: Although statins may provide potential therapeutic pathways for patients with heart failure with preserved ejection fraction (HFpEF), no studies have evaluated statins in combination with standard HF therapy, which would reflect clinical practice more closely. To address this question, we evaluated whether rosuvastatin added to a standard HF therapy provides additional improvement in cardiac structure and function in rats with hypertensive heart failure (SHHF). METHODS AND RESULTS: Two-month-old SHHF rats were randomly assigned to four groups: (i) non-treated SHHF rats; (ii) rosuvastatin-treated SHHF rats; (iii) SHHF rats treated with quinapril plus torasemide plus carvedilol (considered as standard HF therapy); and (iv) SHHF rats treated with the combination of standard HF therapy and rosuvastatin. The administration of a standard anti-hypertensive HF therapy to SHHF rats for 17 months attenuated left ventricular (LV) chamber dilatation, cardiac hypertrophy, fibrosis, and inflammation compared with non-treated SHHF rats. Rosuvastatin alone prevented LV dilatation and cardiac inflammation similar to standard HF therapy-treated SHHF, despite being unable to normalize blood pressure (BP) or influence cardiac hypertrophy. However, and importantly, the addition of rosuvastatin to the standard HF therapy further prevented LV dilatation, preserved cardiac function, and normalized inflammation. CONCLUSION: These data show that the use of rosuvastatin plus a standard HF therapy results in a significant additional improvement in HF and cardiac remodelling in a rat model of HFpEF. These beneficial effects were independent of BP and plasma lipid changes, and seem to be due, at least in part, to decreased myocardial inflammation.

Relationship between plasma levels of soluble CD40L and insulin sensitivity and insulin secretion status in non-diabetic dyslipidemic patients.

OBJECTIVE: We have measured circulating plasma sCD40L as well as the platelet-surface of CD40L and its receptor in a sample of non-diabetic dyslipidemic patients and then evaluated its relationship with the insulin resistance (IR) and insulin secretion (IS) status. DESIGN AND METHODS: Anthropometric measurements, fasting glucose, insulin, lipids, and IR and IS [estimated by the homeostasis model assessment (HOMA)] were assessed in 86 dyslipidemic subjects. Circulating sCD40L were determined by ELISA. By flow cytometry, CD40L, CD40 and P-selectin were evaluated in the platelet-surface. RESULTS: Non-diabetic dyslipidemic IR patients (HOMA-IR>or=3.8) showed higher plasma sCD40L concentrations and a more unfavorable cardiovascular risk profile (higher BMI, waist, fasting insulin and mean triglyceride levels) than dyslipidemic patients with low IS (HOMA beta-cell<98). In a multivariable model, only measures of insulin sensitivity and higher waist remained significantly associated with increased plasma levels of sCD40L. Surface expression of CD40L on platelets decreased significantly and CD40 increased in IR patients, compared with patients with low IS. CONCLUSIONS: IR dyslipidemic patients show increased plasma sCD40L and decreased platelet-membrane CD40L expression compared to dyslipidemic patients with low IS.