RESUMEN
Graft vascular disease (GVD) remains the major limitation to long-term survival after solid organ transplantation. Aortic or carotid allografts in rats have been shown to be useful models because similar changes to those observed in man develop within weeks. Both immunological and non-immunological factors influence the process of GVD and a method that could permit rapid multiple arterial allotransplantation in the rat would be of great value. We performed simultaneous orthotopic aortic and carotid allotransplantations in 25 rats. The vessels were anastomosed using a sleeve technique. No immunosuppression was given. The animals were killed at 15, 30, or 60 days and histological analyses of the grafts were performed. The overall survival rate was 80% and the incidence of technical failure was very low. The histopathological aspect revealed typical progressive GVD. In conclusion, we have developed a new model of simultaneous aortic and carotid transplantation in rats. This model, which incorporates a modification of the sleeve anastomosis, is rapid and yields an easy tool to investigate immunological and non-immunological processes driving GVD.
Asunto(s)
Aorta/trasplante , Arterias Carótidas/trasplante , Cirugía Veterinaria/métodos , Anastomosis Quirúrgica/métodos , Anastomosis Quirúrgica/veterinaria , Animales , Animales no Consanguíneos , Modelos Animales de Enfermedad , Masculino , Ratas , Ratas Wistar , Organismos Libres de Patógenos EspecíficosAsunto(s)
Aorta Abdominal/patología , Aorta Abdominal/trasplante , Arterias Carótidas/patología , Arterias Carótidas/trasplante , Trasplante Homólogo/métodos , Trasplante Isogénico/métodos , Anastomosis Quirúrgica , Animales , Aorta Abdominal/cirugía , Arterias Carótidas/cirugía , Modelos Animales de Enfermedad , Rechazo de Injerto/patología , Homocigoto , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos ICR , Trasplante Homólogo/patología , Trasplante Isogénico/patologíaRESUMEN
Recent studies highlight the existence of an autonomous nuclear polyphosphoinositide metabolism related to cellular proliferation and differentiation. However, only few data document the nuclear production of the putative second messengers, the 3-phosphorylated phosphoinositides, by the phosphoinositide 3-kinase (PI3K). In the present paper, we examine whether GTP-binding proteins can directly modulate 3-phosphorylated phosphoinositide metabolism in membrane-free nuclei isolated from pig aorta smooth muscle cells (VSMCs). In vitro PI3K assays performed without the addition of any exogenous substrates revealed that guanosine 5'-(gamma-thio)triphosphate (GTPgammaS) specifically stimulated the nuclear synthesis of phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P(3)), whereas guanosine 5'-(beta-thio)diphosphate was ineffective. PI3K inhibitors wortmannin and LY294002 prevented GTPgammaS-induced PtdIns(3,4,5)P(3) synthesis. Moreover, pertussis toxin inhibited partially PtdIns(3,4,5)P(3) accumulation, suggesting that nuclear G(i)/G(0) proteins are involved in the activation of PI3K. Immunoblot experiments showed the presence of Galpha(0) proteins in VSMC nuclei. In contrast with previous reports, immunoblots and indirect immunofluorescence failed to detect the p85alpha subunit of the heterodimeric PI3K within VSMC nuclei. By contrast, we have detected the presence of a 117-kDa protein immunologically related to the PI3Kgamma. These results indicate the existence of a G protein-activated PI3K inside VSMC nucleus that might be involved in the control of VSMC proliferation and in the pathogenesis of vascular proliferative disorders.
Asunto(s)
Núcleo Celular/enzimología , Proteínas de Unión al GTP/metabolismo , Músculo Liso Vascular/enzimología , Fosfatidilinositol 3-Quinasas/metabolismo , Activación Enzimática , Técnica del Anticuerpo Fluorescente , Microscopía Electrónica , Músculo Liso Vascular/ultraestructura , Especificidad por SustratoRESUMEN
In addition to their actions on reproductive function, estrogens have important effects on endothelial cells. The present study was designed to evaluate the mechanism(s) by which 17beta-estradiol (E2) promotes endothelial cell proliferation. The potential involvement of vascular endothelial growth factor (VEGF) was investigated by the coadministration of polyclonal anti-VEGF antibody. First, the effect of E2 on the proliferation of cultured foetal bovine aortic endothelial cells (FBAEC) was studied. E2 stimulated this proliferation with an EC50 between 10(-11) and 10(-10) M and this effect was inhibited by the anti-VEGF antibody. The effect of a physiological dose of E2 was then studied in the rat model of carotid injury. After deendothelializing balloon injury, reendothelialization of the denuded surface may influence the growth of the underlying smooth muscle cells. Male Sprague-Dawley rats were castrated and then received E2 from subcutaneously implanted pellets that released 3.2 microg/kg/day. Endothelial regrowth (Evans blue staining) and neointimal thickening were evaluated 2 weeks after the carotid injury. In comparison to the placebo group, E2 increased the extent of reendothelialization (p = 0.0002) and reduced neointimal thickening (p = 0.0007). Anti-VEGF antibody abolished the effect of E2 on reendothelialization as well as on neointimal thickening. Thoracic aorta VEGF content was increased in E2-treated rats compared to control rats. In conclusion, the present study demonstrates that E2 increases endothelial cell proliferation in vitro and reendothelialization in vivo by means of a mechanism dependent on endogenous VEGF. This effect could contribute to the antiatherogenic effect of a physiological dose of E2.
Asunto(s)
Factores de Crecimiento Endotelial/fisiología , Endotelio Vascular/patología , Endotelio Vascular/fisiopatología , Estradiol/farmacología , Linfocinas/fisiología , Mitógenos/farmacología , Cicatrización de Heridas/fisiología , Animales , Aorta Torácica/embriología , Aorta Torácica/lesiones , Aorta Torácica/patología , Aorta Torácica/fisiopatología , Cateterismo/efectos adversos , Bovinos , División Celular/efectos de los fármacos , Células Cultivadas , Factores de Crecimiento Endotelial/biosíntesis , Endotelio Vascular/embriología , Endotelio Vascular/lesiones , Feto , Linfocinas/biosíntesis , Masculino , Ratas , Ratas Sprague-Dawley , Factor A de Crecimiento Endotelial Vascular , Factores de Crecimiento Endotelial VascularRESUMEN
BACKGROUND: Graft arteriosclerosis is a major cause of death after allotransplantation of organs such as the heart or the kidney. Aortic allotransplantation in mice is a useful experimental model to study the mechanisms of this pathology. However, the conventional heterotopic aortic model is limited by a high morbidity and is technically difficult to perform. We developed a new simple method for aortic transplantation in mice. METHODS: The infrarenal aorta from the donor mouse was anastomosed to the recipient's aorta at the same position using a sleeve technique. Orthotopic aortic transplantation was performed in 45 mice, 5 isografts and 40 allografts. No immunosuppression was given, and the mice were killed at day 15 or 30. The graft was examined macroscopically, and several histologic sections were made. RESULTS: The overall survival rate was 78%. The incidence of thrombosis was low (4 cases) compared with previously published series. Histology of aortas revealed typical aspects of rejection in the allografts with a chronic picture at day 30. No significant lesion was observed in isografts. CONCLUSIONS: We have developed a model of orthotopic aortic transplantation in mice. This new model is easy to carry out and has a low incidence of thrombosis, probably because there is no size discrepancy between donor and recipient aortic segment.
Asunto(s)
Aorta Abdominal/trasplante , Arteriosclerosis/etiología , Modelos Animales de Enfermedad , Anastomosis Quirúrgica/métodos , Animales , Aorta Abdominal/patología , Arteriosclerosis/mortalidad , Arteriosclerosis/patología , Femenino , Oclusión de Injerto Vascular/epidemiología , Oclusión de Injerto Vascular/mortalidad , Oclusión de Injerto Vascular/patología , Ratones , Ratones Endogámicos C57BL , Trasplante Homólogo , Trasplante IsogénicoRESUMEN
The microscopic, electron microscopic and immunohistochemical observation of biopsy specimens taken at an early stage, at close and regular intervals (every 4 hours), from open skin wounds created in the pig and the monkey, together with quantitative analysis of the various cell types in the granulation tissue, supports the conception that the activated fibrocyte (fibroblast) originates from the fibrocyte of the wound edges and thus completes some earlier experimental studies. We describe here the various stages of the differentiation of the wound edge fibrocyte into an activated fibrocyte and its proliferation and migration from the edges to the site of the wound. This does not exclude the possibility that local mesenchymal cells take part in the formation of activated fibrocytes. The activated fibrocyte build the collagen of the granulation tissue and then remodel and ensure wound contraction by becoming fibroclasts and myofibroblasts. This article defines the signification of the terms fibrocyte, activated fibrocyte, fibroblast and activated fibroblast.
Asunto(s)
Fibroblastos/fisiología , Piel/lesiones , Cicatrización de Heridas , Heridas y Lesiones/patología , Heridas y Lesiones/fisiopatología , Animales , Femenino , Fibroblastos/patología , Fibroblastos/ultraestructura , Microscopía Electrónica , Ovariectomía , Piel/patología , Piel/ultraestructura , PorcinosRESUMEN
Benfluorex is a clinical lipid-lowering agent with antihyperglycemic properties. The effect of long-term oral treatment (10 mg/kg/day for 7.5 months) on carbohydrate and lipid metabolism and aortic morphology was investigated in 24 insulin-resistant sand rats receiving a standard laboratory diet supplemented with cholesterol (2%). Untreated controls (n=34) developed impaired glucose tolerance, hyperinsulinemia, hypertriglyceridemia, hypercholesterolemia and elevated plasma LDL- and VLDL-cholesterol, positively correlated with the proportion of the thoracic aorta displaying oil red O-positive atherosclerosis; ultrastructural examination showed intimal lipid deposits, foam cells, polymorph infiltrates and fibrosis. Benfluorex-treated animals showed significant decreases in glucose intolerance, hyperinsulinemia, hypertriglyceridemia, hypercholesterolemia, and plasma LDL- and VLDL-cholesterol, with no evidence of aortic atheroma. The metabolic benefits of benfluorex may protect against the long-term development of atherosclerosis in the insulin-resistant dyslipidemic syndrome.
Asunto(s)
Fenfluramina/análogos & derivados , Hiperlipidemias/metabolismo , Hipolipemiantes/farmacología , Resistencia a la Insulina , Hígado/efectos de los fármacos , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Aorta/efectos de los fármacos , Aorta/patología , Arteriosclerosis/prevención & control , Peso Corporal/efectos de los fármacos , Colesterol en la Dieta/efectos adversos , Femenino , Fenfluramina/farmacología , Gerbillinae , Prueba de Tolerancia a la Glucosa , Hiperlipidemias/etiología , Lípidos/sangre , Hígado/química , Masculino , RatasRESUMEN
Proliferation of vascular smooth muscle cells (SMC) is a hallmark in the pathogenesis of atherosclerotic lesions. Mildly oxidized low density lipoproteins (UV-oxLDL), which are mitogenic to cultured AG-08133A SMC, activate the sphingomyelin (SM)-ceramide pathway. We report here the following. (i) UV-oxLDL elicited a biphasic and sustained activation of MBP kinase activity, phosphorylation and nuclear translocation of p44/42 mitogen-activated protein kinase (MAPK), and [3H]thymidine incorporation, which were inhibited by PD-098059, a MAPK kinase inhibitor. (ii) The use of preconditioned media (from SMC pre-activated by UV-oxLDL) transferred to native SMC and blocking antibodies against growth factors suggest that UV-oxLDL-induced activation of MAPK and [3H]thymidine incorporation seem to be independent of any autocrine secretion of growth factors. (iii) UV-oxLDL-induced activation of a neutral sphingomyelinase, SM hydrolysis, ceramide production, and [3H]thymidine incorporation were inhibited by two serine-protease inhibitors (serpins), suggesting that a serpin-sensitive proteolytic pathway is involved in the activation of the SM-ceramide signaling pathway. (iv) UV-oxLDL-induced MAPK activation and [3H]thymidine incorporation were mimicked by ceramide generated in the plasma membrane by bacterial sphingomyelinase treatment or by addition of the permeant C2-ceramide. Serpins did not inhibit the MAPK activation and [3H]thymidine incorporation induced by C2-ceramide, indicating that activation of the MAPK and [3H]thymidine incorporation is subsequent to the stimulation of the SM-ceramide pathway. Taken together, these data suggest that mitogenic concentrations of UV-oxLDL are able to stimulate the SM-ceramide pathway through a protease-dependent mechanism and activate p44/42 MAPK, leading to proliferation of vascular SMC.
Asunto(s)
Proteínas Quinasas Dependientes de Calcio-Calmodulina/metabolismo , División Celular/fisiología , Lipoproteínas LDL/farmacología , Músculo Liso Vascular/efectos de los fármacos , Esfingomielinas/fisiología , Animales , Bovinos , Células Cultivadas , Activación Enzimática , Cinética , Músculo Liso Vascular/citología , Oxidación-Reducción , Esfingomielina Fosfodiesterasa/metabolismo , Esfingomielinas/metabolismo , Timidina/metabolismoRESUMEN
BACKGROUND: The cytokines interleukin 1 (IL-1) and tumor necrosis factor (TNF) are secreted by the different cell populations of the vascular wall and have been suggested to promote atherosclerosis. METHODS AND RESULTS: Their respective roles in fatty-streak formation in apolipoprotein E-deficient mice were investigated by use of IL-1 receptor antagonist and TNF binding protein. Estradiol-17beta was used as a positive control. Blocking TNF seemed to be active in female animals but not in males. IL-1 receptor antagonist was as effective as or more effective than estradiol in both sexes. CONCLUSIONS: IL-1 plays a crucial role in the initial step of the atherosclerotic process in this animal model, and blocking the activity of this cytokine should be considered as a therapeutic possibility.
Asunto(s)
Apolipoproteínas E/deficiencia , Arteriosclerosis/etiología , Proteínas Portadoras/fisiología , Receptores del Factor de Necrosis Tumoral , Sialoglicoproteínas/fisiología , Animales , Arteriosclerosis/patología , Proteínas Portadoras/administración & dosificación , Proteínas Portadoras/farmacología , Estradiol/farmacología , Femenino , Proteína Antagonista del Receptor de Interleucina 1 , Metabolismo de los Lípidos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Receptores Tipo I de Factores de Necrosis Tumoral , Sialoglicoproteínas/administración & dosificación , Sialoglicoproteínas/farmacología , Receptores Señuelo del Factor de Necrosis TumoralRESUMEN
Human lymphocytes derived from the peripheral blood of a healthy woman were transfected with a plasmid carrying the simian virus 40 (SV40) large T antigen. The successfully transformed cells contained SV40 large T DNA and were negative for Epstein-Barr virus (EBV) and human T-cell leukaemia virus (HTLV)-1 genomes. The immortalized cell line was assigned to the T-lymphocyte lineage on the basis of morphological, immunological and cytochemical criteria. While the cells expressed CD1a and CD4 at the cell surface, the CD3 complex was solely intracytoplasmic. Immunoprecipitation studies indicated that these cells lacked T-cell receptor (TCR) alpha-chains but not beta-chains. They were negative for activation markers such as CD25, CD69 and major histocompatibility (MHC) class II molecules. In addition, the transformed cells exhibited a complete growth independency towards interleukin-2 (IL-2). However, after phorbol ester stimulation, CD25 and CD69 markers were expressed and IL-2 was secreted. This new human immortalized T-lymphocytic cell line, which is cell-surface TCR/CD3-negative, may be useful as an in vitro model for studying TCR/CD3 assembly, expression and signal transduction.
Asunto(s)
Antígenos Transformadores de Poliomavirus/metabolismo , Complejo Receptor-CD3 del Antígeno de Linfocito T/biosíntesis , Linfocitos T/metabolismo , Antígenos Transformadores de Poliomavirus/genética , División Celular , Línea Celular , Transformación Celular Viral , Femenino , Humanos , Cariotipificación , Fenotipo , Linfocitos T/citologíaRESUMEN
The reality of the atheroprotective effect of estrogens is still a matter of debate, and its unknown mechanisms could involve favorable changes in blood lipids and lipoproteins and/or direct action at the level of the arterial wall. We used the recently developed animal model of atherosclerosis constituted by apolipoprotein E-deficient mice in an attempt to clarify these issues. Male and female animals, fed a low-fat chow diet, were treated with increasing doses of 17 beta-estradiol (E2) after castration and compared with testosterone treated and uncastrated (intact) animals. Total serum cholesterol, LDL-cholesterol, and HDL-cholesterol concentrations decreased under E2 treatment in each sex and were weakly correlated with lesion area. However, a highly significant correlation between lesion area and serum E2 levels also suggested a direct action of E2 on cells of the vascular wall. A dose-response curve analysis revealed that these activities were sex-dependent, with females being nearly twice as sensitive to E2 as males. It also revealed that the atheroprotective activity was recruited at higher E2 concentrations than those needed by other E2 target tissues such as uterus or functions such as apoA-1 and LDL production and/or clearance rates.
Asunto(s)
Apolipoproteínas E/deficiencia , Arteriosclerosis/prevención & control , Estradiol/uso terapéutico , Animales , Aorta/patología , Enfermedades de la Aorta/patología , Enfermedades de la Aorta/prevención & control , Apolipoproteínas E/genética , Arteriosclerosis/sangre , Arteriosclerosis/genética , Arteriosclerosis/patología , Peso Corporal/efectos de los fármacos , Castración , Colesterol/sangre , Grasas de la Dieta/administración & dosificación , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Estradiol/sangre , Estradiol/farmacología , Femenino , Lipoproteínas/sangre , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Miocardio/patología , Caracteres Sexuales , Método Simple Ciego , Testosterona/sangre , Testosterona/farmacologíaRESUMEN
Human endothelial cells (ECs) grown under standard conditions are able to generate a basal level of oxygen free radicals and induce progressive oxidation of LDLs. Inhibition of cell-mediated LDL oxidation by superoxide dismutase, EDTA, or desferrioxamine implicates a role for superoxide anion and/or transition metals in this process. The potential role of the mitochondrion was investigated by inducing mitochondrial deenergization by selective photosensitization or the addition of inhibitors of the mitochondrial respiratory chain. Mitochondria of human cultured ECs were selectively damaged by photosensitization of cells labeled with the mitochondrion-selective fluorescent dye 2-(4-dimethylaminostyryl)-1-methylpyridinium iodide under conditions that induced only low levels of toxicity during the time of the experiment. Photosensitized ECs exhibited severe mitochondrial dysfunction, as suggested by the defect in mitochondrial uptake of the mitochondrion-selective fluorescent dyes [rhodamine 123 and 2-(4-dimethylaminostyryl)-1-methylpyridinium iodide] and morphological alterations as shown by transmission electron microscopy. In mitochondria-photosensitized cells, superoxide anion generation was strongly decreased, as was LDL oxidation and the subsequent cytotoxicity. When ECs were incubated with the mitochondrial respiratory-chain inhibitors antimycin A or rotenone or with the carbonylcyanide-m-chlorophenylhydrazone uncoupler rhodamine 123, uptake and subcellular distribution were altered, and concomitantly superoxide anion production and LDL oxidation were strongly decreased. In conclusion, these data suggest that mitochondrial function is required, directly or indirectly, for the production of superoxide anion and the subsequent LDL oxidation by human vascular ECs.
Asunto(s)
LDL-Colesterol/metabolismo , Endotelio/citología , Endotelio/metabolismo , Mitocondrias/fisiología , Carbonil Cianuro m-Clorofenil Hidrazona/farmacología , Línea Celular , Deferoxamina/farmacología , Inhibidores Enzimáticos/farmacología , Colorantes Fluorescentes/farmacología , Humanos , Ionóforos/farmacología , Luz , Manitol/farmacología , Mitocondrias/efectos de la radiación , NG-Nitroarginina Metil Éster/farmacología , Oxidación-Reducción , Superóxido Dismutasa/antagonistas & inhibidores , Superóxidos/metabolismo , omega-N-Metilarginina/farmacologíaRESUMEN
Skin fibroblasts from patients with Farber disease (acid ceramidase deficiency) and from two siblings of the only known family affected with prosaposin deficiency were transformed by transfection with a plasmid carrying the SV40 large T antigen. The prosaposin-deficient transformed cell lines conserved their original metabolic defects, and in particular they were free of detectable immunoreactivity when using anti-saposin B and anti-saposin C antisera. Ultrastructurally, the cells contained heterogeneous lysosomal storage products. As found for their parental cell lines, the SV40-transformed fibroblasts exhibited deficient in vitro activities of lysosomal ceramidase and beta-galactosylceramidase, but a normal activity of acid sphingomyelinase. As observed for SV40-transformed fibroblasts from Farber disease, degradation of radioactive glucosylceramide or low density lipoprotein-associated radiolabelled sphingomyelin by the prosaposin-deficient cells in situ showed a clear impairment in the turnover of lysosomal ceramide. Ceramide storage in prosaposin-deficient cells was also demonstrated by ceramide mass determination. In contrast to acid ceramidase deficient cells, both the accumulation of ceramide and the reduced in vitro activity of acid ceramidase in cells from prosaposin deficiency could be corrected by addition of purified saposin D. The data confirm that prosaposin is required for lysosomal ceramide degradation, but not for sphingomyelin turnover. The SV40-transformed fibroblasts will be useful for pathophysiological studies on human prosaposin deficiency.
Asunto(s)
Amidohidrolasas/deficiencia , Amidohidrolasas/genética , Transformación Celular Viral/genética , Glicoproteínas/deficiencia , Glicoproteínas/genética , Virus 40 de los Simios/genética , Ceramidasa Ácida , Amidohidrolasas/metabolismo , Antígenos Transformadores de Poliomavirus/genética , Línea Celular Transformada , Ceramidasas , Feto , Fibroblastos/química , Fibroblastos/metabolismo , Fibroblastos/ultraestructura , Glicoproteínas/metabolismo , Humanos , Inmunodifusión , Enfermedades por Almacenamiento Lisosomal/genética , Enfermedades por Almacenamiento Lisosomal/metabolismo , Precursores de Proteínas/deficiencia , Precursores de Proteínas/genética , Precursores de Proteínas/metabolismo , SaposinasRESUMEN
Oxidized LDLs are thought to play a central role in atherogenesis. Among their wide variety of biological properties, oxidized LDLs exhibit a cytotoxic effect on cultured vascular cells. Toxic doses of mildly oxidized LDLs elicited massive apoptosis in both primary and immortalized cultures of endothelial cells as shown by characteristic morphological and biochemical changes. Cytoplasmic and nucleic modifications (eg, chromatin condensation and nucleus fragmentation) were visualized by using electron and fluorescence microscopy of intact cells labeled by the fluorescent DNA probe SYTO-11. DNA fragmentation was quantified by ultracentrifugation of chromatin fragments, evaluated in situ by using the TUNEL (Terminal transferase-mediated dUTP-biotin nick end labeling) procedure, and visualized by electrophoresis of radiolabeled DNA fragments showing the characteristic apoptotic ladder. Apoptotic cells became rapidly detached and underwent postapoptotic necrosis that led to cell disintegration. Apoptosis was subsequent to a sustained and delayed peak of cytosolic calcium. Both the calcium peak and apoptosis were blocked by chelating the extracellular calcium with EGTA or by inhibiting the calcium influx by the calcium-channel blockers nifedipine and nisoldipine, thus suggesting that the apoptotic process induced by oxidized LDLs is clearly calcium dependent. Aurintricarboxylic acid, an inhibitor of endonucleases, also blocked the apoptotic process without blocking the calcium peak. These results suggest that toxic doses of mildly oxidized LDLs induce massive apoptosis of endothelial cells through a calcium-dependent mechanism and that this apoptotic process can be prevented by inhibiting the rise of cytosolic calcium or by inhibiting cellular endonucleases by aurintricarboxylic acid.
Asunto(s)
Apoptosis/efectos de los fármacos , Ácido Aurintricarboxílico/farmacología , Calcio/fisiología , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Lipoproteínas LDL/farmacología , Células Cultivadas , Humanos , Lipoproteínas LDL/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-bcl-2/metabolismoRESUMEN
Oxidized low density lipoprotein (LDL) is thought to play a major role in atherogenesis. Atherosclerotic arteries exhibit structural changes associated with profound alterations in vascular tone that are potentially involved in arterial spasm and ischemic heart disease. We report here the role of oxidized LDL in the retraction of vascular smooth muscle cells. Mildly oxidized LDL elicited a broad and sustained peak in cytosolic calcium concentration ([Ca2+]i) in cultured arterial smooth muscle cells. Concomitant with the [Ca2+]i rise, oxidized LDL evoked a sustained and intense retraction of smooth muscle cells, as shown by the changes in cross-sectional area of single cells. Cell retraction was dependent on time, the concentration of oxidized LDL, and the level of LDL oxidation (native LDL induced neither a significant [Ca2+]i rise nor cell retraction). Oxidized LDL but not native LDL also elicited a delayed (12 +/- 2 hours) and sustained (14 +/- 2 hours) increase in isometric tension in deendothelialized arterial rings only, thus suggesting a protective role of intact endothelium. When triggered by nontoxic doses of oxidized LDL, retraction of cultured cells and the contractile response of aortic rings was reversible, whereas with higher (toxic) doses (> or = 200 micrograms apoB/mL), cell retraction was irreversible and led progressively to detachment and cell death. Cell retraction can be prevented in three ways: (1) by inhibiting LDL oxidation with supplements of antioxidants (indirect inhibition); (2) by blocking the pathogenic intracellular signaling elicited by oxidized LDL (direct inhibition), eg, by inhibiting calcium influx with EGTA or the calcium channel blocker nisoldipine or by blocking intracellular signaling (at a still-unknown step) by the lipophilic antioxidant alpha-tocopherol; and (3) by directly inhibiting myosin light chain kinase by 1-(5-chloronaphthalene-1-sulfonyl)-1H-hexahydro-1, 4-diazepine. In conclusion, oxidized LDL evoked a sustained and intense calcium-dependent retraction of cultured smooth muscle cell, which can be prevented by inhibiting LDL oxidation or by blocking the intracellular signaling induced by oxidized LDL.
Asunto(s)
Calcio/metabolismo , Lipoproteínas LDL/farmacología , Contracción Muscular/efectos de los fármacos , Músculo Liso Vascular/fisiología , Animales , Bovinos , Células Cultivadas , Peroxidación de Lípido , Lipoproteínas LDL/metabolismoRESUMEN
Sand rats fed a hypercholesterolaemic diet containing 0.01% of the anti-thyroid agent 2-mercapto-1-imidazole develop preatheromatous lesions similar to those found in humans, in addition to obesity and insulin resistance. The effects of a nutritional supplement rich in essential fatty acids and garlic extract (Arterodiet) on the appearance and evolution of the lesions were studied. Treatment with this nutritional supplement significantly decreased circulating triglycerides and low-density lipoprotein (LDL)-cholesterol levels but did not alter plasma insulin or glucose levels. Intra-arterial cholesterol levels were also decreased by the treatment which resulted in a normalisation of the atherosclerotic lesions in these animals.
Asunto(s)
Fenómenos Fisiológicos Nutricionales de los Animales , Arteriosclerosis/prevención & control , Gerbillinae/fisiología , Animales , Glucemia/metabolismo , LDL-Colesterol/metabolismo , Dieta , Semivida , Hormonas/sangre , Insulina/sangre , Masculino , Triglicéridos/metabolismoRESUMEN
This study compares the ability of intracoronary ultrasound (ICUS) to identify thrombus by means of actual criteria, with the histologic studies of tissues removed by directional atherectomy in patients treated previously with thrombolytic therapy. Coronary angiography and intravascular ultrasound imaging were performed before atherectomy in 34 patients who had received intravenous thrombolytic therapy for acute myocardial infarction a mean of 6 days before. The lesion morphology and the percentage of stenosis were defined on the angiogram. The ultrasound characteristics of the narrowing were described as intraluminal thrombus, mural thrombus, mixed plaque, and dense plaque. Thirty patients were studied. Thrombus was suspected in 8 patients on angiography. By ICUS, the presence of thrombus was predicted in 21 patients. Histologic studies of excised tissues found thrombus in 20 of the 30 patients. When ICUS was compared with histology, the true-positive rate was 80% and the false-positive rate was 50%; the true-negative rate was 50% and the false-negative rate was 20%. The correlation between observers was high. These observations suggest that ICUS may be useful in identifying fresh thrombus. The findings of this study help to confirm the criteria for diagnosing intraluminal thrombus by ICUS imaging.
Asunto(s)
Aterectomía Coronaria , Trombosis Coronaria/diagnóstico por imagen , Trombosis Coronaria/patología , Ultrasonografía Intervencional , Adulto , Anciano , Angiografía Coronaria , Trombosis Coronaria/etiología , Vasos Coronarios/diagnóstico por imagen , Vasos Coronarios/patología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Infarto del Miocardio/complicaciones , Sensibilidad y EspecificidadRESUMEN
This study focuses on the fortuitous discovery of an atypical atherosclerotic lesion in four of 49 male adult cynomolgus monkeys (macacus fascicularis) which were maintained for a long time at a high level of hypercholesterolemia, and in seven of 19 female cynomolgus monkeys examined from the second to the 24th week of hypercholesterolemic diet: this lesion was in formation or already mature during this period of diet. This atypical lesion was formed by a collagen and elastic network surrounding synthetic smooth muscle cells without fibrofatty or fibrous plaques. Lipids were occasionally seen in the inner intima. The lesion appeared early (from the third week of diet). Once established, its morphology did not change. It became more extensive, but was not complicated by lipid overload in spite of prolonged, permanent hypercholesterolemia. This response to hypercholesterolemia is interesting because the activity of the smooth muscle cells differs from that observed in the classic lesion: they intervene earlier, their replication is very marked and rapid, their elastin secretion is greater and remains constant over time, and their phagocytic properties are reduced. This experimental study examines the installation and the maintenance of this lesion and raises the problem of its origin.
Asunto(s)
Enfermedades de la Aorta/patología , Arteriosclerosis/patología , Hipercolesterolemia/patología , Músculo Liso Vascular/fisiopatología , Animales , Aorta Abdominal/patología , Aorta Torácica/patología , Enfermedades de la Aorta/sangre , Enfermedades de la Aorta/etiología , Arteriosclerosis/sangre , Arteriosclerosis/etiología , División Celular , Colágeno/análisis , Vasos Coronarios/patología , Dieta Aterogénica , Elastina/análisis , Femenino , Hipercolesterolemia/sangre , Hipercolesterolemia/complicaciones , Lípidos/sangre , Macaca fascicularis , Macrófagos/ultraestructura , Masculino , Músculo Liso Vascular/ultraestructuraRESUMEN
We have investigated the role of low-density lipoprotein (LDL) oxidation in the proliferative effect of LDLs on cultured bovine aortic smooth-muscle cells and compared it with their effect on bovine aortic endothelial cells. The following conclusions were reached. (1) Non-toxic doses of mildly oxidized LDLs elicit a proliferative effect on smooth-muscle cells significantly higher than that of native LDLs or lipoprotein-depleted serum. The proliferative effect is dependent on time (relatively slow), dose (high doses are cytotoxic) and the level of LDL oxidation. (2) The proliferative effect on smooth-muscle cells is counterbalanced at high concentrations of mildly oxidized LDLs (or at high oxidation levels) by their cytotoxic effect. (3) The same dose of mildly oxidized LDLs exhibits no proliferative effect on endothelial cells but rather a cytotoxic one. Endothelial cells may therefore be intrinsically more susceptible to the cytotoxic effect of mildly oxidized LDLs than are smooth-muscle cells. (4) The proliferative effect of native LDLs on smooth-muscle cells results (at least in part) from cell-induced LDL oxidation during cell culture as suggested by (i) the progressive LDL oxidation over the 3 days of contact between LDLs and smooth-muscle cells and (ii) the concomitant inhibition of LDL oxidation and proliferative effect by butylated hydroxytoluene. The hypothetical mechanisms and potential involvement in atherogenesis are discussed.
Asunto(s)
Lipoproteínas LDL/farmacología , Músculo Liso Vascular/efectos de los fármacos , Animales , Bovinos , División Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Humanos , Lipoproteínas LDL/química , Lipoproteínas LDL/toxicidad , Músculo Liso Vascular/citología , Oxidación-ReducciónRESUMEN
Bearing in mind earlier studies which established a link between arteriosclerosis and mineral loss, or fragility of the bones, and also our recent study showing that patients with arterial disorders of the lower limbs also suffered from osteoporosis, we carried out a histological study of the number and appearance of the intraosseous vessels and trabecular bone volume in the femoral heads of patients undergoing surgery for either fracture of the femoral neck or osteoarthritis of the hip. The number of thick-walled vessels, arterioles or arterial capillaries was significantly diminished in the femoral heads of patients with fractures of the femoral neck (p = 0.007). In addition, in the latter patients, arteriosclerotic vascular lesions (rupture of the internal elastic lamina, medial thickening and fibrosis) were more frequent than in patients with osteoarthritis of hip. The possibility that, through chronic ischemia, arteriosclerosis may lead to disturbance of bone remodelling and loss of the mechanical properties of bone has not been contradicted by these findings.