RESUMEN
Experimental ischemia-reperfusion models have shown that 3-hydroxy-3methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, statins, have cardioprotective effects. SAMIT (Statin Acute Myocardial Infarction Trial) is a multicenter prospective open randomized trial, designed to evaluate the effects of statin treatment from the earliest stage on cardioprotection in patients with acute myocardial infarction (AMI). Patients were randomly assigned to receive atorvastatin (initial dose of 40 mg at admission followed by the maintenance dose of 10 mg/day for 30 days) or not (control), and then immediately underwent percutaneous coronary intervention (PCI) for the culprit lesion. The primary endpoints were infarct size and left ventricular function. The secondary endpoints were major adverse cardiac and cerebrovascular events (MACCE) and various biomarkers. There were no significant differences in baseline characteristics between 2 groups of the statin treatment group and the control group. The left ventricular ejection fraction increased at 6 months after the onset of AMI, compared with the baseline level in the atorvastatin group (P < 0.05), while it did not change in the control group. Although there were no significant differences in the MACCE, the changes in the levels of angiopoietin-like protein 2 (ANGPTL2) (P < 0.05), and glyceraldehyde-derived advanced glycation end-products, (TAGE) (P < 0.01) were suppressed at 2 weeks in the atorvastatin group, compared with the control group. Statin therapy started early after the onset reduced the levels of ANGPTL2 and TAGE, and thus, might have cardioprotective effects in patients with AMI.
Asunto(s)
Angioplastia Coronaria con Balón , Angiopoyetinas/sangre , Atorvastatina/administración & dosificación , Productos Finales de Glicación Avanzada/sangre , Inhibidores de Hidroximetilglutaril-CoA Reductasas/administración & dosificación , Infarto del Miocardio/terapia , Enfermedad Aguda , Anciano , Proteína 2 Similar a la Angiopoyetina , Proteínas Similares a la Angiopoyetina , Femenino , Humanos , Japón , Masculino , Persona de Mediana Edad , Estrés Oxidativo/efectos de los fármacos , Estudios Prospectivos , Resultado del Tratamiento , Función Ventricular Izquierda/efectos de los fármacosRESUMEN
BACKGROUND: The purpose of this study was to elucidate the effects of glimepiride on the levels of biomarkers related to cardiovascular regulation in patients with type 2 diabetes mellitus. METHODS AND RESULTS: Thirty-four patients with type 2 diabetes received glimepiride for 24 weeks. Significant decreases in the levels of glyceraldehyde-derived advanced glycation end products, (glycer-AGE: toxic AGE), eotaxin and fibroblast growth factor (FGF)-2 were recognized after the administration of glimepiride. Moreover, there were trends for there to be increases in the levels of granulocyte-colony stimulating factor (G-CSF) and granulocyte macrophage-colony stimulating factor (GM-CSF), and decreases in the levels of fractalkine, soluble CD40 ligand (sCD40L), macrophage inflammatory protein (MIP)-ß, vascular endothelial growth factor (VEGF) and soluble receptor for AGE (sRAGE). CONCLUSIONS: Glimepiride may have potent anti-oxidative, anti-inflammatory and angiogenic properties and it may potentially repair tissue damage by decreasing the levels of toxic AGE and increasing colony-stimulating factors.
Asunto(s)
Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Índice Glucémico/fisiología , Hipoglucemiantes/uso terapéutico , Compuestos de Sulfonilurea/uso terapéutico , Anciano , Femenino , Índice Glucémico/efectos de los fármacos , Humanos , Hipoglucemiantes/farmacología , Masculino , Persona de Mediana Edad , Compuestos de Sulfonilurea/farmacología , Resultado del TratamientoRESUMEN
Familial hypercholesterolemia (FH) is an autosomal codominant disease characterized by high concentrations of proatherogenic lipoproteins secondary to deficiency in low-density lipoprotein (LDL) receptor. We reported recently the use of in situ stem cell therapy of human adipose tissue-derived multilineage progenitor cells (hADMPCs) in lowering serum total cholesterol in the homozygous Watanabe heritable hyperlipidemic (WHHL) rabbits, an animal model of homozygous FH. Here we demonstrate that pravastatin, an HMG-CoA reductase inhibitor, augmented the cholesterol-lowering effect of transplanted hADMPCs and enhanced LDL clearance in homozygous WHHL rabbit. The results suggest the potential beneficial effects of in situ stem cell therapy in concert with appropriately selected pharmaceutical agents, in regenerative medicine.
Asunto(s)
Tejido Adiposo/citología , Colesterol/sangre , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacología , Hiperlipoproteinemia Tipo II/cirugía , Pravastatina/farmacología , Trasplante de Células Madre/métodos , Células Madre/efectos de los fármacos , Adulto , Animales , Linaje de la Célula/efectos de los fármacos , Femenino , Humanos , Persona de Mediana Edad , Conejos , Células Madre/fisiología , Adulto JovenRESUMEN
Tumor nests in lung squamous cell carcinoma (LUSC) have a hierarchical structure resembling squamous epithelium. The nests consist of basal-like cells on the periphery and layers of keratinocyte-like cells that differentiate towards the center of the nest, forming keratin pearls. Reproducing this spatial heterogeneity in in vitro models would be useful for understanding the biology of LUSC. Here, we established a three-dimensional (3D) culture model with a squamous epithelial structure using LUSC cell lines PLR327F-LD41 and MCC001F, established in-house. When PLR327F-LD41 cells were cultured in a mixture of Matrigel and collagen I, they generated 3D colonies (designated cancer organoids, or COs) with involucrin (IVL)-positive keratinizing cells in the center (IVLinner COs). COs with uniform size were generated by seeding PLR327F-LD41 cells in a form of small cell aggregates. Since Notch signaling induces the differentiation of squamous epithelium, we confirmed the effect of γ-secretase inhibitor in inhibiting Notch signaling in IVLinner COs. Surprisingly, γ-secretase inhibitor did not block induction of IVL-positive cells; however, cells residing between the CK5-positive basal-like layer and IVL-positive layer decreased significantly. Thus, our 3D culture model with uniform size and structure promises to be a useful tool for elucidating the biology of LUSC and for screening drug-candidates.
Asunto(s)
Carcinoma de Células Escamosas/metabolismo , Técnicas de Cultivo de Célula , Neoplasias Pulmonares/metabolismo , Modelos Biológicos , Animales , Carcinoma de Células Escamosas/patología , Línea Celular Tumoral , Femenino , Humanos , Neoplasias Pulmonares/patología , RatonesRESUMEN
Accumulating evidence suggests that inflammation as well as oxidative stress play essential roles in atherogenesis, progression of atherosclerosis, and plaque instability and rupture. Recent studies on available anti-hypertensive agents have focused on their anti-atherosclerotic effects over and above their blood pressure lowering action. These studies have included investigations on several types of calcium channel blockers, with several investigations indicating that a dihydropiridine-based calcium channel blocker, azelnidipine, developed in Japan, has unique anti-oxidative properties. An anti-inflammatory effect of azelnidipine has, however, yet to be established and therefore we carried out a series of in vivo and in vitro studies to investigate this possibility. This was achieved by measuring inflammatory and oxidative stress markers in 16 high risk hypertensive patients administered 16mg/day of azelnidipine. After 4 weeks of treatment, serum levels of hsCRP, IL-6, and IL-8 and urinary 8-OHdG were decreased significantly, despite blood pressure remaining unchanged. Cultures of human mononuclear leukocytes collected from six healthy volunteers showed 100 nM of azelnidipine caused significant inhibition of formyl-methyonyl leucyl phenylalanine (fMLP)-induced production of IL-8. Taken together, these results suggest that azelnidipine has anti-inflammatory effects independent of its anti-hypertensive action. As leukocytes do not possess voltage-operated calcium channels, the effect of azelnidipine in these cells appears to occur independently of an L-type calcium channel antagonizing effect.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Ácido Azetidinocarboxílico/análogos & derivados , Bloqueadores de los Canales de Calcio/farmacología , Dihidropiridinas/farmacología , 8-Hidroxi-2'-Desoxicoguanosina , Adulto , Anciano , Anciano de 80 o más Años , Aterosclerosis/prevención & control , Ácido Azetidinocarboxílico/farmacología , Biomarcadores/sangre , Biomarcadores/orina , Proteína C-Reactiva/metabolismo , Desoxiguanosina/análogos & derivados , Desoxiguanosina/orina , Femenino , Humanos , Hipertensión/sangre , Hipertensión/tratamiento farmacológico , Hipertensión/orina , Técnicas In Vitro , Mediadores de Inflamación/sangre , Interleucina-6/sangre , Interleucina-8/sangre , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/metabolismo , Masculino , Persona de Mediana Edad , Estrés Oxidativo/efectos de los fármacosRESUMEN
Type 1 diabetes mellitus is caused by autoimmune destruction of insulin-producing beta cells. The major obstacle to transplantation of insulin-producing cells to cure the disease is the limited source of these cells. To overcome this problem, we describe here a multistep protocol for generation of insulin-producing islet-like clusters from human adipose tissue-derived stromal cells (ADSCs). Analysis using reverse transcription polymerase chain reaction detected enhanced expression of various pancreatic genes during the differentiation of ADSCs. Immunofluorescence analysis revealed functional similarities between cells derived from ADSCs and pancreatic islet cells, i.e., the presence of insulin- and C-peptide-coexpressing cells in the clusters and glucagon expression on the cell surface. The glucose challenge tests revealed the production of insulin, and such production was regulated via physiological signaling pathways. Our insulin-producing cells derived from ADSCs could be potentially used for cell therapy of type 1 diabetes mellitus.
Asunto(s)
Tejido Adiposo/citología , Transdiferenciación Celular , Células Secretoras de Insulina/citología , Adulto , Femenino , Expresión Génica , Humanos , Insulina/metabolismo , Secreción de Insulina , Células Secretoras de Insulina/metabolismo , Masculino , Persona de Mediana Edad , Hormonas Pancreáticas/metabolismo , Células del EstromaRESUMEN
The goal of the study was to examine if allogenic mesenchymal stem cell (MSC) transplantation is a useful therapy for acute myocardial infarction (AMI). Buffer (control; group C, n=41), MSCs of male ACI rats (allogenic; group A, n=38, 5 x 10(6)), or MSCs of male LEW rats (syngenic; group S, n=40, 5 x 10(6)) were injected into the scar 15 min after myocardial infarction in female LEW rats. After 28 days, fractional left ventricular shortening significantly increased in groups A (21.3+/-1.7%, P=0.0467) and S (23.2+/-1.9%, P=0.0140), compared to group C (17.1+/-0.9%). Fibrosis in groups A and S was significantly lower. Quantitative PCR of the male-specific sry gene showed disappearance of donor cells within 28 days (5195+/-1975 cells). Secretion of vascular endothelial growth factor (VEGF) by MSCs was enhanced under hypoxic conditions in vitro. In groups A and S, the plasma VEGF concentration, VEGF level, and capillary density in recipient hearts increased after 28 days. Flow cytometry revealed the absence of B7 signal molecules on MSCs. A mixed lymphocyte reaction showed that ACI MSCs failed to stimulate proliferation of LEW lymphocytes. After 1 day after cell transplantation, transient increases in interleukin-1 beta and monocyte chemoattractant protein-1 in recipient hearts were enhanced in group A, with macrophage infiltration at the injection site. T cells remained at the level of normal tissue in all groups. We conclude that allogenic MSC transplantation therapy is useful for AMI. The donor MSCs disappear rapidly, but become a trigger of VEGF paracrine effect, without induction of immune rejection.
Asunto(s)
Trasplante de Células Madre Mesenquimatosas , Infarto del Miocardio/terapia , Animales , Recuento de Células , Supervivencia Celular , Modelos Animales de Enfermedad , Fibrosis Endomiocárdica/diagnóstico por imagen , Fibrosis Endomiocárdica/patología , Femenino , Infarto del Miocardio/inducido químicamente , Infarto del Miocardio/inmunología , Infarto del Miocardio/patología , Miocardio/patología , Neovascularización Patológica , Comunicación Paracrina , Ratas , Ratas Endogámicas Lew , Trasplante Homólogo , Ultrasonografía , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
BACKGROUND: Recently, accumulating evidence has indicated that bone marrow-derived stem cells are capable of differentiating into vascular cells. It has been hypothesized that the inflammatory response after vascular injury triggers the mobilization of endothelial and smooth muscle progenitor cells from bone marrow. METHODS AND RESULTS: We measured circulating CD34-positive mononuclear cells, activation of integrin Mac-1 on the surface of neutrophils, and plasma granulocyte-colony stimulating factor levels in 40 patients undergoing coronary stenting. After bare-metal stenting, CD34-positive cells increased, reaching a maximum on day 7 after stenting. The maximum change compared with baseline before stenting was more striking in patients with restenosis than without restenosis (332+/-108% versus 148+/-49%; P<0.05). In contrast, CD34-positive cells decreased after sirolimus-eluting stenting (72+/-21% on day 7). The change in CD34-positive cells on day 7 relative to baseline was closely correlated with that in activated Mac-1 at 48 hours (R=0.52, P<0.01) and that in granulocyte-colony stimulating factor levels at 24 hours (R=0.42, P<0.05). Cell culture assay on day 7 showed that mononuclear cells differentiated into CD31-positive endothelium-like cells after bare-metal stenting. In patients with restenosis, mononuclear cells differentiating into alpha-smooth muscle actin-positive smooth muscle-like cells also were observed. Implantation of sirolimus-eluting stents suppressed both types of differentiation. CONCLUSIONS: Stent implantation may induce differentiation of bone marrow cells into endothelial or smooth muscle cells. Endothelial cells may participate in reendothelialization, a protective reaction against vascular injury, whereas smooth muscle cells may participate in neointimal thickening and restenosis. Sirolimus-eluting stents appear to inhibit the mobilization and differentiation of bone marrow cells.
Asunto(s)
Antígenos CD34/sangre , Células de la Médula Ósea/citología , Movimiento Celular , Reestenosis Coronaria/sangre , Stents/efectos adversos , Anciano , Células de la Médula Ósea/fisiología , Cateterismo Cardíaco/efectos adversos , Movimiento Celular/fisiología , Enfermedad de la Arteria Coronaria/sangre , Enfermedad de la Arteria Coronaria/terapia , Reestenosis Coronaria/etiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Factores de TiempoRESUMEN
AIM: Recently, calcium channel blockers (CCBs) have been reported to reduce atherosclerosis with anti-inflammatory or antiatherosclerotic effects in vivo. It is well established that monocytes and macrophages play important roles in promoting atherosclerosis. However, the effects of CCBs on macrophage activation remain unclear. The aim of this study was to evaluate the effects of azelnidipine, a dihydropyridine L-type CCB, on the activation of macrophages and to clarify the mechanisms of the effects of CCBs on atherosclerosis. METHODS: THP-1 monocytes, a human leukemic cell line, were stimulated with 50 ng/mL of phorbol-12-myristate-13-acetate (PMA) 1 h after pretreatment with 10 µM azelnidipine or dimethyl sulfoxide (DMSO), and harvested. RESULTS: Azelnidipine blocked the expression of intercellular adhesion molecule-1 quantified by FACS analysis. The expression levels of Apo E and MMP9, which are markers of macrophage differentiation, were inhibited by azelnidipine as evaluated by quantitative RT-PCR. The level of LOX-1 mRNA, a scavenger receptor, was also reduced significantly by pretreatment with 10 µM azelnidipine. Azelnidipine also lowered the uptake of acetylated LDL. The expression of the L-type calcium channel Cav1.2 was 10-fold higher after 24 h of PMA stimulation. A knockdown of the CACNA1C gene, which encodes Cav1.2 protein in humans, with siRNA blocked the effect of reducing adhesion by azelnidipine, indicating that the effects of azelnidipine on macrophage differentiation were expressed through the CACNA1C gene. CONCLUSION: Our results suggest that azelnidipine has potent antiatherosclerotic properties by inhibition of macrophage activation through Cav1.2.
Asunto(s)
Ácido Azetidinocarboxílico/análogos & derivados , Canales de Calcio Tipo L/metabolismo , Adhesión Celular/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Dihidropiridinas/farmacología , Activación de Macrófagos/efectos de los fármacos , Macrófagos/citología , Ácido Azetidinocarboxílico/farmacología , Células Cultivadas , Humanos , Macrófagos/efectos de los fármacos , Macrófagos/metabolismoRESUMEN
Adiponectin is an adipocyte-specific secretory protein that is highly and specifically expressed in adipose tissue, and low plasma levels of adiponectin are associated with coronary artery disease (CAD). It has been suggested that high molecular weight (HMW) adiponectin is more important for vascular protection than total amount of adiponectin. To establish the clinical relevance of HMW adiponectin, we measured its serum levels in 149 patients with CAD. The levels were lower in vasospastic angina pectoris (3.4 +/- 2.4 microg/ml, p <0.01), stable angina pectoris (3.3 +/- 2.6 microg/ml, p <0.001), and healed myocardial infarction (3.8 +/- 2.9 microg/ml, p <0.01) than chest pain syndrome (controls) (6.6 +/- 5.4 microg/ml). The levels were also lower in multivessel CAD (3.4 +/- 2.4 microg/dl) compared with single vessel CAD (4.2 +/- 2.7 microg/ml, p <0.05) or no organic stenosis (5.1 +/- 3.5 microg/ml, p <0.01). In univariate analysis, diabetes mellitus (p = 0.03), insulin resistance (p = 0.06), high-sensitivity C-reactive protein levels (p = 0.0012), and low HMW adiponectin levels (p = 0.0001) predicted cardiovascular events during 7 years of follow-up. However, multivariate analysis showed that only HMW adiponectin levels were an independent predictor of cardiovascular events (relative risk 2.79, 95% confidence interval 1.49 to 5.24, p = 0.0014). In conclusion, serum HMW adiponectin levels may serve as a predictor of future cardiovascular events in patients with CAD as well as a marker for severity of CAD.
Asunto(s)
Adiponectina/sangre , Biomarcadores/sangre , Enfermedad Coronaria/sangre , Anciano , Angiografía Coronaria , Enfermedad Coronaria/diagnóstico , Enfermedad Coronaria/mortalidad , Progresión de la Enfermedad , Ensayo de Inmunoadsorción Enzimática , Femenino , Estudios de Seguimiento , Humanos , Japón/epidemiología , Masculino , Persona de Mediana Edad , Peso Molecular , Pronóstico , Ventriculografía con Radionúclidos , Estudios Retrospectivos , Factores de Riesgo , Índice de Severidad de la Enfermedad , Tasa de Supervivencia/tendencias , Factores de TiempoRESUMEN
Hypercholesterolemia is a well-known risk factor for a wide range of diseases in developed countries. Here, we report that mice lacking functional LDLR (low density lipoprotein receptor), an animal model of human familial hypercholesterolemia, show circadian abnormalities. In free running behavioral experiments in constant darkness, these mice showed a prolonged active phase and distinctly bimodal rhythms. Even when the circadian rhythms were entrained by light and dark cycles, these mice showed a significant attenuation of behavioral onset intensity at the start of the dark period. Further, we hypothesized that the combination of hypercholesterolemia and circadian abnormalities may affect cardiovascular disease progression. To examine this possibility, we generated LDLR-deficient mice with impaired circadian rhythms by simultaneously introducing a mutation into Period2, a core clock gene, and found that these mice showed a significant enlargement of artery plaque area with an increase in inflammatory cytokine IL-6 levels. These results suggest that circadian dysfunction may be associated with the development or progression of cardiovascular diseases.
Asunto(s)
Enfermedades Cardiovasculares/etiología , Enfermedades Cardiovasculares/metabolismo , Ritmo Circadiano , Hipercolesterolemia/sangre , Hipercolesterolemia/complicaciones , Animales , Enfermedades Cardiovasculares/patología , Ritmo Circadiano/genética , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Masculino , Ratones , Ratones Noqueados , Mutación , Proteínas Circadianas Period/genética , Fotoperiodo , Receptores de LDL/genética , Factores de RiesgoRESUMEN
BACKGROUND: Manganese superoxide dismutase (MnSOD) is an important antioxidant enzyme affected in heart/muscle-specific MnSOD-deficient mice (H/M-SOD2-/-), which develop progressive congestive heart failure and exhibit pathology typical of dilated cardiomyopathy. METHODS: In this study we investigated the beneficial effects of epigallocatechin gallate (EGCG) on the cardiac remodeling and telomere biology in H/M-SOD2-/- mice. H/M-SOD2-/- mice were divided into three groups: those receiving normal drinking water (KO), a low dose of EGCG (L: 10mg/L), and a high dose of EGCG (H: 100mg/L) beginning at eight weeks of age and lasting for eight weeks. RESULTS: The mice in the KO group exhibited significantly dilated cardiac remodeling with reduced contractility, which was prevented by the administration of EGCG. Although the mortality of KO mice was about 50% at 16 weeks of age, the mice that received EGCG had a high survival rate. The cardiac dilatation with reduced cardiac contraction in KO mice was prevented by EGCG treatment. The levels of myocardial oxidative stress and free fatty acids were lower in the group treated with EGCG compared with the KO group. The increased expression of nitric oxide synthase 2, nitrotyrosine, fatty acid synthase, Toll-like receptor 4, and Sirt1 in the KO mice were prevented by EGCG treatment. The shortening of the telomere length, decreased telomerase activity in KO mice were also prevented by EGCG. CONCLUSIONS: H/M-SOD2-/- mice receiving EGCG have a lower mortality rate and exhibit less inflammation and a better preserved cardiac function and telomere biology.
Asunto(s)
Antioxidantes/administración & dosificación , Catequina/análogos & derivados , Insuficiencia Cardíaca/tratamiento farmacológico , Animales , Catequina/administración & dosificación , Modelos Animales de Enfermedad , Ácido Graso Sintasas/efectos de los fármacos , Ácidos Grasos no Esterificados/metabolismo , Ratones , Ratones Noqueados , Miocardio/metabolismo , Óxido Nítrico Sintasa de Tipo II/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Sirtuina 1/efectos de los fármacos , Superóxido Dismutasa/genética , Telomerasa/efectos de los fármacos , Acortamiento del Telómero/efectos de los fármacos , Receptor Toll-Like 4/efectos de los fármacos , Tirosina/análogos & derivados , Tirosina/efectos de los fármacos , Remodelación Ventricular/efectos de los fármacosRESUMEN
BACKGROUND: An excessive inflammatory response after myocardial infarction (MI) increases myocardial injury. The toll-like receptor (TLR)-4 is activated by the recognition of endogenous ligands and is proinflammatory when there is myocardial tissue injury. The apoptosis inhibitor of the macrophage (AIM) is known to provoke an efflux of saturated free fatty acids (FFA) due to lipolysis, which causes inflammation via the TLR-4 pathway. Therefore, this study investigated the hypothesis that AIM causes a proinflammatory response after MI. METHODS AND RESULTS: The left anterior descending coronary artery was ligated to induce MI in both AIM-knockout (AIM(-/-)) and wild-type (WT) mice. After 3 days, the inflammatory response from activation of the TLR-4/NFκB pathway was assessed, and infarct size was measured by staining with triphenyltetrazolium chloride. In addition, left ventricular remodeling was examined after 28 days. Although the area at risk was similar between AIM(-/-) and WT mice, the infarct size was significantly smaller in AIM(-/-) mice (P=0.02). The heart weight-to-body weight ratio and myocardial fibrosis were also decreased in the AIM(-/-) mice, and the 28-day survival rate was improved (P<0.01). With the reduction of plasma FFA in AIM(-/-) mice, myocardial IRAK4 and NFκB activity were decreased (all P<0.05). Moreover, there was a reduction in myeloperoxidase activity and inducible nitric oxide synthase as part of the inflammatory response (P<0.01, P=0.03, respectively). Furthermore, NFκB DNA-binding activation via TLR-4, neutrophil infiltration, and inflammatory mediators were decreased in AIM(-/-) mice. CONCLUSIONS: The deletion of AIM reduced the inflammatory response and infarct size and improved survival after myocardial infarction.
Asunto(s)
Proteínas Reguladoras de la Apoptosis/genética , Proteínas Reguladoras de la Apoptosis/fisiología , Inflamación/fisiopatología , Infarto del Miocardio/fisiopatología , Receptores Inmunológicos/genética , Receptores Inmunológicos/fisiología , Animales , Western Blotting , Modelos Animales de Enfermedad , Ecocardiografía , Ácidos Grasos no Esterificados/sangre , Hemodinámica/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Infarto del Miocardio/diagnóstico por imagen , Infarto del Miocardio/patología , FN-kappa B/sangre , Receptores DepuradoresRESUMEN
BACKGROUND AND AIMS: Pentraxin-3 (PTX3) reportedly has protective roles in atherosclerosis and myocardial infarction, and is a useful biomarker of vascular inflammation. However, the detailed functions of PTX3 in inflammation are yet to be elucidated. This study aimed to investigate the function of PTX3 in macrophages. METHODS: PMA-treated THP-1 cell line (THP-1 macrophage) and monocyte-derived human primary macrophages were treated with recombinant PTX3. Cytokine and chemokine levels in the THP-1 culture medium were measured as well as monocyte chemoattractant protein (MCP-1) concentrations in the Raw 264.7 cell culture medium. PTX3-silenced apoptotic macrophages (THP-1 cell line) were generated to investigate the roles of PTX3 in phagocytosis. RESULTS: In the presence of PTX3, macrophage interleukin-1ß (IL-1ß), tumor necrosis factor-alpha (TNF-α) and MCP-1 levels were reduced significantly (-39%, P=0.007; -21%, P=0.008; and -67%, P=0.0003, respectively), whilst activated transforming growth factor-ß (TGF-ß) was detected in the THP-1 macrophages (P=0.0004). Additionally, PTX3 induced Akt phosphorylation and reduced nuclear factor-kappa B (NF-κB) activation by 35% (P=0.002), which was induced by TNF-α in THP-1 macrophages. Furthermore, silencing of PTX3 in apoptotic cells resulted in increased macrophage binding, elevated expression rate of HLA-DR (+30%, P=0.015) and CD86 (+204%, P=0.004) positive cells, and induction of IL-1ß (+36%, P=0.024) production. Conversely, adding recombinant PTX3 to macrophages reduced CD86 and HLA-DR expression in a dose-dependent manner. CONCLUSIONS: We identified PTX3 as a novel regulator of macrophage activity, and this function suggests that PTX3 acts to resolve inflammation.
RESUMEN
BACKGROUND: Obstructive sleep apnea (OSA) is characterized by repetitive intermittent hypoxia and reoxygenation during sleep with elevated oxidative stress and promotes the development of atherosclerosis, as demonstrated by vascular dysfunction and chronic inflammation. An increased neutrophil to lymphocyte ratio (NLR) has been recognized to be a novel inflammatory biomarker for systemic inflammation. OBJECTIVES: We evaluated whether the NLR reflects the severity of OSA and if continuous positive airway pressure (CPAP) treatment ameliorates the endothelial function and NLR in patients with OSA. METHODS: We enrolled 95 patients with suspected OSA and 29 patients who received CPAP therapy for 3 months. We evaluated the number of endothelial progenitor cells (EPCs) and NLR, the levels of nitric oxide (NOx) and asymmetric dimethylarginine (ADMA), and the endothelial function according to the flow-mediated dilatation (FMD) before and after CPAP treatment. RESULTS: The levels of apnea-hypopnea index demonstrated an inverse relationship with the FMD and a positive relationship with the NLR. Moreover, NLR is an independent factor suggested for the presence of severe OSA. CPAP therapy increased the levels of EPC and NOx and decreased the level of ADMA. CPAP treatment also improved the FMD and decreased the NLR. CONCLUSIONS: NLR and endothelial dysfunction significantly correlates with the severity of OSA and FMD and other biochemical parameters improved and NLR decreased significantly after CPAP treatment.
Asunto(s)
Endotelio Vascular/fisiopatología , Linfocitos/metabolismo , Neutrófilos/metabolismo , Índice de Severidad de la Enfermedad , Apnea Obstructiva del Sueño/sangre , Adulto , Anciano , Arginina/análogos & derivados , Biomarcadores/sangre , Presión de las Vías Aéreas Positiva Contínua , Células Progenitoras Endoteliales/metabolismo , Femenino , Humanos , Recuento de Linfocitos , Masculino , Persona de Mediana Edad , Óxido Nítrico/sangre , Estudios Prospectivos , Apnea Obstructiva del Sueño/fisiopatología , Apnea Obstructiva del Sueño/terapiaRESUMEN
BACKGROUND: The mechanism involved in the spontaneous acceptance of liver allografts in some rat strain combinations remains unclear. Immunoregulatory NKR-P1TCRalphabetaT (NKT) cells primarily produce IL-4 and IFN-gamma, and enhance the polarization of immune responses to Th2 and Th1, respectively. The aim of this study was to clarify the role of graft-derived NKT cells in inducing the spontaneous acceptance of rat orthotopic liver transplantation (OLTx) METHODS: The experimental groups were divided as follows: Group 1, BN to LEW "low responder (acceptor)" combination; Group 2, DA to LEW "high responder (rejector)" combination; naïve BN (Group 3) or LEW recipients (Group 4) with liver allografts from irradiated BN donors. The recipients had liver allografts from irradiated donors reconstituted from the following cell populations 24 hr before harvesting, spleen cells (SPCs, Group 5), IgSPCs (Group 6), IgNKR-P1SPCs (Group 7), and IgTCRabSPCs (Group 8) RESULTS: In Group 1, the percent of graft-derived NKT cells harvested on day 7 posttransplant were significantly higher than in Group 2. In the case of BN liver allografts that had been irradiated and reconstituted with cell populations including NKT cells (Groups 5 and 6), the mean graft survival (MST) was extended to 39.2+/-5.7 and 38.8+/-8.0 days, respectively. In contrast, when NKT cells were excluded (Groups 7 and 8), the grafts were acutely rejected within MST of 17.8+/-4.0 and 18.8+/-7.7 days, respectively. The concentrations of IL-10 and TGF-beta, but not IL-4 in IgGICs culture supernatants were predominant in the acceptor, whereas those with IFN-gamma predominated in the rejector. CONCLUSIONS: Graft-derived NKT cells might be responsible for spontaneous acceptance in the rat OLTx.
Asunto(s)
Antígenos de Superficie/análisis , Lectinas Tipo C/análisis , Trasplante de Hígado/inmunología , Receptores de Antígenos de Linfocitos T alfa-beta/análisis , Linfocitos T/inmunología , Animales , Citocinas/análisis , Citometría de Flujo , Rechazo de Injerto/inmunología , Supervivencia de Injerto/inmunología , Trasplante de Hígado/fisiología , Masculino , Modelos Animales , Subfamilia B de Receptores Similares a Lectina de Células NK , Ratas , Ratas Endogámicas BN , Ratas Endogámicas Lew , Bazo/inmunología , Trasplante Homólogo/inmunología , Trasplante Homólogo/fisiología , Irradiación Corporal TotalRESUMEN
BACKGROUND: In terms of the temporal relationship between pancreas transplantation (PTx) and reversal of diabetic ocular complications, it has been difficult but important to determine a "point of no return." Thus, it is of great clinical interest to evaluate the efficacy of PTx on diabetic ocular complications. METHODS: A spontaneous type 2 diabetic model of Spontaneously Diabetic Torii (SDT; RT1) rats was used in the present study, and syngeneic PTx was performed. RESULTS: In the control SDT rats that received no treatment, hyperglycemia (>250 mg/dL) was developed from 25.2+/-3.9 weeks of age. Lens opacity was observed in all rats at 15 weeks after the onset of diabetes. Fluorescein angiography and immunohistochemistry detected the nonperfusion area and neovascularization in the retina at 5 weeks of diabetes. Daily insulin treatment could not prevent or reverse the ocular changes in our experiment. Fluorescein filling defect of the retinal vessels was observed at 10 weeks of diabetes. However, in the PTx rats, normoglycemia was achieved at all experimental time points. Diabetic cataract and retinopathy could have been prevented and improved if PTx had been performed at 5 weeks, but not at 10 weeks after the onset of diabetes. With PTx treatment, an inhibition of angiogenesis in the retina at 5 weeks after the onset of diabetes was demonstrated by immunohistochemistry. CONCLUSIONS: Our results indicate that the potential use of the SDT rat for diabetes study and the positive effect of PTx performed before the "point of no return" could prevent and cure diabetic ocular complications.
Asunto(s)
Catarata/prevención & control , Diabetes Mellitus Tipo 2/cirugía , Retinopatía Diabética/prevención & control , Trasplante de Páncreas , Animales , Glucemia , Catarata/epidemiología , Catarata/patología , Diabetes Mellitus Tipo 2/epidemiología , Diabetes Mellitus Tipo 2/patología , Retinopatía Diabética/epidemiología , Retinopatía Diabética/patología , Modelos Animales de Enfermedad , Incidencia , Masculino , Páncreas/patología , Páncreas/cirugía , Ratas , Ratas MutantesRESUMEN
Adenovirus-mediated CTLA4Ig gene transfer has been reported to enhance graft survival in several rodent transplantation models. In this study, we investigated the efficacy of ex vivo and systemic transfer of the CTLA4Ig gene by adenoviral vectors in pancreatic islet allo-transplantation. Islet grafts from BN rats were transplanted to chemically induced diabetic LEW rats. First, ex vivo CTLA4Ig gene transfer into isolated islets was performed prior to transplantation. Survival of transduced grafts under the kidney capsule was slightly prolonged (8.6+/-1.3 days) compared with survival of untransduced grafts (6.7+/-1.2 days); when combined with a short course of FK506, graft survival was further extended (32.6+/-10.7 days vs. 13.7+/-1.0 days with FK506 alone). Secondly, systemic gene transfer was accomplished by intravenous administration immediately after the transplantation procedure. In these animals, islet grafts under the kidney capsule survived longer (15.2+/-3.3 days) than in controls (6.7+/-1.2 days), and when FK506 was administered perioperatively, all the islet grafts survived for more than 100 days. In systemically transduced recipients, the survival of islet grafts transplanted into the liver was not significantly different from that of the grafts placed under the kidney capsule. In order to examine organ-specific immunogenicity, heterotopic BN cardiac grafts were transplanted to LEW rats intra-abdominally, with the virus transferred systemically as in the islet model. In contrast to the islet grafts, all the cardiac grafts were accepted for longer than 100 days, even without FK506 therapy. Finally, the LEW recipients with long-surviving islet or cardiac grafts were re-transplanted with islet grafts from the same donor strain (BN) on day 100. The second islet grafts survived longer than 100 days in half of the cardiac recipients, but consistently failed in the islet recipients. We conclude that in this transplant model, CTLA4Ig gene transfer and FK506 treatment synergistically improved islet graft survival, systemic transfer of the gene was more effective than ex vivo transfer to the islets, and donor-specific tolerance could not be achieved for islet transplantation but was achieved for cardiac transplantation.
Asunto(s)
Adenoviridae , Técnicas de Transferencia de Gen , Vectores Genéticos , Inmunoconjugados/genética , Abatacept , Animales , Inmunoconjugados/sangre , Inmunosupresores/farmacología , Islotes Pancreáticos/patología , Trasplante de Islotes Pancreáticos/inmunología , Ratas , Tacrolimus/farmacología , Tolerancia al Trasplante/efectos de los fármacos , Tolerancia al Trasplante/inmunologíaRESUMEN
AIM: The number of hepatitis A cases in Japan as well as in other developed countries has been progressively decreasing during the last several years. There is no universal hepatitis A vaccination program in Japan, and a hepatitis A virus (HAV) epidemic in Japan is not unlikely. In 2011, a hepatitis A outbreak associated with a revolving sushi bar occurred in Chiba, Japan. We aimed to analyze this outbreak. METHODS: Twenty-seven patients associated with this outbreak were admitted to the National Hospital Organization Chiba Medical Center. Molecular epidemiologic investigations were conducted. RESULTS: Twenty-six of the 27 patients had gone to the same revolving sushi bar, and then clinical symptoms appeared. HAV RNA was detected by reverse transcription polymerase chain reaction in 23 of the 27 (85.1%) patients whose sera had tested positive for anti-HAV immunoglobulin M. All isolates from this outbreak were clustered within subgenotype IA, displaying 100% sequence homology with each other in 232 bp from all 23 patients. All isolates belong to the IA-1 sublineage, which is endemic to Japan. CONCLUSION: A revolving sushi bar was associated with a hepatitis A outbreak, and molecular epidemiological investigations proved useful.
RESUMEN
OBJECTIVE: Glucagon-like peptide 1 (GLP-1), one of the incretin hormones, has been reported to increase positive inotropic activity in cardiac myocytes and protect against myocardial injury. However, the effects upon endothelial cells and the mechanisms involved are not fully understood. We assessed the hypothesis that GLP-1 has protective effects against inflammation and oxidative stress on human endothelial cells. METHODS AND RESULTS: The effects of the GLP-1 analog liraglutide upon TNF-α-induced injury of the human umbilical vein endothelial cells (HUVECs) were evaluated. First, ROS induced by TNF-α was measured by staining with CM-H(2)DCFDA. Intracellular ROS production of HUVECs was significantly decreased in a dose-dependent manner until 30 nM while liraglutide inhibited the induction of gp91(phox) and p22(phox), subunit of NADPH oxidase, by TNF-α. In addition, protein levels of SOD-2, catalase and GPx were significantly increased by liraglutide. Second, rapid translocation of PKC-α into the membrane following TNF-α was evident. Liraglutide significantly inhibited this very rapid TNF-α-induced translocation of PKC-α into membrane at 2.5 min. Third, liraglutide significantly inhibited NF-κB activation and upregulated I-κB family while phosphorylation of IKK-α/ß, which is upstream of NF-κB signaling, was also downregulated after 15 min of TNF-α treatment. Finally, liraglutide inhibited apoptosis of HUVEC and expression of Pentraxin-3 induced by TNF-α. CONCLUSION: Liraglutide exerts marked anti-oxidative and anti-inflammatory effects on endothelial cells with inhibition of PKC-α, NADPH oxidase, NF-κB signaling and upregulation of protective anti-oxidative enzymes.