RESUMEN
Novel atherosclerosis models are needed to guide clinical therapy. Here, we report an in vitro model of early atherosclerosis by fabricating and perfusing multi-layer arteriole-scale human tissue-engineered blood vessels (TEBVs) by plastic compression. TEBVs maintain mechanical strength, vasoactivity, and nitric oxide (NO) production for at least 4 weeks. Perfusion of TEBVs at a physiological shear stress with enzyme-modified low-density-lipoprotein (eLDL) with or without TNFα promotes monocyte accumulation, reduces vasoactivity, alters NO production, which leads to endothelial cell activation, monocyte accumulation, foam cell formation and expression of pro-inflammatory cytokines. Removing eLDL leads to recovery of vasoactivity, but not loss of foam cells or recovery of permeability, while pretreatment with lovastatin or the P2Y11 inhibitor NF157 reduces monocyte accumulation and blocks foam cell formation. Perfusion with blood leads to increased monocyte adhesion. This atherosclerosis model can identify the role of drugs on specific vascular functions that cannot be assessed in vivo.
Asunto(s)
Arteriolas/fisiopatología , Aterosclerosis/fisiopatología , Arteriolas/química , Arteriolas/citología , Aterosclerosis/genética , Aterosclerosis/metabolismo , Fenómenos Biomecánicos , Adhesión Celular , Proliferación Celular , Células Cultivadas , Células Espumosas/citología , Células Espumosas/metabolismo , Humanos , Lipoproteínas LDL/metabolismo , Modelos Biológicos , Monocitos/citología , Monocitos/metabolismo , Óxido Nítrico/metabolismo , Ingeniería de Tejidos , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
The microvascular architecture of the spleen plays an important role in the immunological function of this organ. The different types of vessels are related to different reticular cells each with their own immunomodulatory functions. The present study describes an immunohistochemical and morphometric analysis of the various types of vessels in 21 human autopsy non-pathological splenic samples. On an area of 785,656.37⯵m2 for each sample, we classified and quantified the type and number of vascular structures, each according to their morphology and immunohistochemical profile, and obtained the ratios between them. The distribution of trabecular vessels and the characteristics of the venules are reviewed. In our material the so-called "cavernous perimarginal sinus" (anatomical structure previously described by Schmidt et al., 1988) was observed and interpreted as a curvilinear venule shaped by the follicle in contact with the trabecular vein. Our material comprised 261 trabeculae (containing 269 arterial sections and 508 venous sections), 30,621 CD34+ capillaries, 7739 CD271+ sheathed capillaries, 2588 CD169+ sheathed capillaries, and 31,124 CD8+ sinusoids. The total area (TA) (14,765,714.88⯵m2) occupied by the sinusoidal sections of the 21 cases was much higher than the TA of the capillary sections (1,700,269.83⯵m2). Similarly, the TA (651,985⯵m2) occupied by the sections of the trabecular veins was much higher than the TA of the trabecular arteries (88,594⯵m2). The total number of CD34+ capillaries and of sinusoids CD8+ was similar for the sum of the 21 cases, nevertheless there were large differences in each case. Statistically the hypothesis that the number of capillaries and sinusoids are present with the same frequency is discarded. In view of the absence of a numerical correlation between capillaries and sinusoids, we postulate that very possibly the arterial and the venous vascular trees are two anatomically independent structures separated by the splenic cords. We believe that this is the first work where splenic microvascularization is simultaneously approached from a morphometric and immunohistochemical point of view.
Asunto(s)
Microvasos/anatomía & histología , Microvasos/química , Bazo/irrigación sanguínea , Actinas/inmunología , Adapaleno/inmunología , Antígenos CD34/inmunología , Arteriolas/anatomía & histología , Arteriolas/química , Autopsia , Antígenos CD8/inmunología , Moléculas de Adhesión Celular , Patologia Forense , Humanos , Inmunoglobulinas/inmunología , Inmunohistoquímica , Mucoproteínas/inmunología , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/inmunología , Lectina 1 Similar a Ig de Unión al Ácido Siálico/inmunología , Bazo/anatomía & histología , Arteria Esplénica/anatomía & histología , Arteria Esplénica/químicaRESUMEN
AIM: The present study was performed to examine the clinicopathological significance of hyaline deposits in the smooth muscle of the interlobular artery (interlobular hyaline arteriopathy [IHA]) in renal allografts. METHODS: Tissue specimens that included the interlobular artery from biopsies performed from January 2012 to December 2015, as well as specimens from biopsies performed ≥1 year after living kidney transplantation were analyzed. Biopsies of recipients with new-onset diabetes mellitus after transplantation were excluded, as well as those of recipients who had undergone transplantation because of diabetic nephropathy. Arteriolopathy was evaluated using the aah score determined by the Banff 2007 classification. RESULTS: In total, 51 specimens with IHA lesions were identified among 381 biopsies obtained from 243 recipients performed ≥1 year after kidney transplantation. Among these 51 biopsies, 18 specimens had a score of aah3, 29 had a score of aah2, and four had a score of aah1. The incidence of IHA lesions was 3.6% at ≥1 to <4 years, 18.5% at ≥4 to <8 years, and 54.1% at ≥8 years. Older kidney grafts exhibited more IHA lesions. Among the biopsy specimens obtained ≥8 years after transplantation, no significant differences in the recipient or donor age, duration after transplantation, or prevalence of hypertension were observed between the IHA and non-IHA groups. The aah scores were significantly higher in the IHA group ≥8 years after transplantation as determined by the mean score test (P < 0.01). CONCLUSION: IHA in renal allografts is associated with severe arteriolopathy.
Asunto(s)
Hialina , Trasplante de Riñón/efectos adversos , Riñón/irrigación sanguínea , Músculo Liso Vascular/química , Enfermedades Vasculares/metabolismo , Aloinjertos , Arteriolas/química , Arteriolas/patología , Biopsia , Humanos , Incidencia , Trasplante de Riñón/métodos , Donadores Vivos , Músculo Liso Vascular/patología , Prevalencia , Arteria Renal/química , Arteria Renal/patología , Índice de Severidad de la Enfermedad , Factores de Tiempo , Tokio/epidemiología , Resultado del Tratamiento , Enfermedades Vasculares/epidemiología , Enfermedades Vasculares/patologíaRESUMEN
AIM: Arteriolar hyalinosis (AH) is a common lesion in allograft biopsies taken following kidney transplantation. Recent studies have shown that severe AH may predict transplant outcomes and provide information about previous exposure to certain drugs, such as calcineurin inhibitors (CNI). However, the incidence of AH as a direct result of diabetic nephropathy (DN) after kidney transplantation has not been fully evaluated. This study aimed to assess the impact of primary DN on the development of AH lesions in patients who underwent kidney transplantation. METHODS: Eighty-three patients who underwent living-donor kidney transplantation between April 2005 and June 2015 were enrolled in this study. A total of 33 patients had DN prior to transplantation. Allograft biopsies were scored according to the Banff classification, and the relationship between the individual histological lesions and clinical baseline data was assessed. RESULTS: At early biopsy (3-12 months), there were no differences in the rates of AH lesions between the DN group and the non-DN group (ah ≥ 1: 37% vs. 41.3%, P = 0.719; aah ≥ 1: 14.8% vs. 6.5%; P = 0.453). However, there were significant differences between the groups in biopsies taken more than 3 years after the transplant (ah ≥ 2: 83.3% vs. 36.8%, P = 0.013; aah ≥ 2: 66.7% vs. 21.1%, P = 0.011). Multivariable analysis showed that both the length of time after transplantation and the presence of DN were independent risk factors for ah ≥ 2 (odds ratio [OR]: 2.55, 95% confidence interval [CI]: 1.47-19.54, P = 0.011) and aah ≥ 2 (OR: 7.55, 95% CI: 1.49-38.33, P = 0.015). CONCLUSION: This is the first report showing that the presence of primary DN disease contributes to the development of severe AH late in the course after kidney allografts.
Asunto(s)
Arteriolas/química , Nefropatías Diabéticas/epidemiología , Hialina , Trasplante de Riñón/efectos adversos , Riñón/irrigación sanguínea , Enfermedades Vasculares/metabolismo , Adulto , Anciano , Aloinjertos , Arteriolas/patología , Biopsia , Distribución de Chi-Cuadrado , Nefropatías Diabéticas/patología , Femenino , Humanos , Incidencia , Japón/epidemiología , Trasplante de Riñón/métodos , Donadores Vivos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Análisis Multivariante , Oportunidad Relativa , Factores de Riesgo , Factores de Tiempo , Resultado del Tratamiento , Enfermedades Vasculares/epidemiología , Enfermedades Vasculares/patologíaRESUMEN
BACKGROUND: Maintenance of brain circulation during shock is sufficient to prevent subcortical injury but the cerebral cortex is not spared. This suggests area-specific regulation of cerebral blood flow (CBF) during hemorrhage. METHODS: Cortical and subcortical CBF were continuously measured during blood loss (≤50%) and subsequent reperfusion using laser Doppler flowmetry. Blood gases, mean arterial blood pressure (MABP), heart rate and renal blood flow were also monitored. Urapidil was used for α1A-adrenergic receptor blockade in dosages, which did not modify the MABP-response to blood loss. Western blot and quantitative reverse transcription polymerase chain reactions were used to determine adrenergic receptor expression in brain arterioles. RESULTS: During hypovolemia subcortical CBF was maintained at 81 ± 6% of baseline, whereas cortical CBF decreased to 40 ± 4% (p < 0.001). Reperfusion led to peak CBFs of about 70% above baseline in both brain regions. α1A-Adrenergic blockade massively reduced subcortical CBF during hemorrhage and reperfusion, and prevented hyperperfusion during reperfusion in the cortex. α1A-mRNA expression was significantly higher in the cortex, whereas α1D-mRNA expression was higher in the subcortex (p < 0.001). CONCLUSIONS: α1-Adrenergic receptors are critical for perfusion redistribution: activity of the α1A-receptor subtype is a prerequisite for redistribution of CBF, whereas the α1D-receptor subtype may determine the magnitude of redistribution responses.
Asunto(s)
Corteza Cerebral/irrigación sanguínea , Circulación Cerebrovascular , Hipovolemia/fisiopatología , Receptores Adrenérgicos alfa 1/metabolismo , Choque/fisiopatología , Agonistas de Receptores Adrenérgicos alfa 1/farmacología , Análisis de Varianza , Animales , Presión Arterial , Arteriolas/química , Arteriolas/metabolismo , Análisis de los Gases de la Sangre , Modelos Animales de Enfermedad , Femenino , Frecuencia Cardíaca , Hemorragia/fisiopatología , Piperazinas/farmacología , Circulación Renal , Reperfusión , OvinosRESUMEN
INTRODUCTION: Renin synthesis and release is the rate-limiting step in the renin-angiotensin system, because cyclic adenosine monophosphate (cAMP) has been identified as dominant pathway for renin gene expression, and cAMP response element-binding protein (CREB) is found in the human and mouse renin promoter. This study aimed to evaluate the role of CREB in expression of the renin gene. MATERIALS AND METHODS: We created conditional deletion of CREB in mice with low-sodium diet, specifically in renin cells of the kidney. To assess the effect of CREB on renin expression, immunostaining of renin was used in samples from wild-type mice and mice with gene knock-down of CREB. Cyclic AMP response element-binding-protein-binding protein (CBP) and p300 were measured in cultured renin cells of the mice, and RNA detection was done with real-time polymerase chain reaction. RESULTS: With low-sodium diet, renin was expressed along the whole wall of the afferent glomerular arterioles in wild-type mice, while there was no increase or even decrease in renin expression in CREB-specific deletion mice; RNA level of renin in cultured cells decreased by 50% with single knock-down of CREB, CBP, or p300, and decreased 70% with triple knock-down of CREB, CBP, and p300. CONCLUSIONS: This study found that CREB was important for renin synthesis and the role of CREB can be achieved through the recruitment of co-activators CBP and p300.
Asunto(s)
Arteriolas/química , Proteína de Unión a CREB/genética , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/genética , Proteína p300 Asociada a E1A/genética , Aparato Yuxtaglomerular/química , ARN Mensajero/análisis , Renina/genética , Animales , Proteína de Unión a CREB/análisis , Células Cultivadas , Colforsina/farmacología , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/análisis , Regulación hacia Abajo/genética , Proteína p300 Asociada a E1A/análisis , Expresión Génica , Técnicas de Silenciamiento del Gen , Inmunohistoquímica , Aparato Yuxtaglomerular/irrigación sanguínea , Aparato Yuxtaglomerular/citología , Ratones , Músculo Liso Vascular/citología , Miocitos del Músculo Liso/química , ARN Interferente Pequeño/genética , Renina/análisis , Sodio en la Dieta/administración & dosificación , TransfecciónRESUMEN
Complement activation has a major role in thrombotic microangiopathy (TMA), a disorder that can occur in a variety of clinical conditions. Promising results of recent trials with terminal complement-inhibiting drugs call for biomarkers identifying patients who might benefit from this treatment. The primary aim of this study was to determine the prevalence and localization of complement factor C4d in kidneys of patients with TMA. The secondary aims were to determine which complement pathways lead to C4d deposition and to determine whether complement activation results in deposition of the terminal complement complex. We examined 42 renal sections with histologically confirmed TMA obtained from a heterogeneous patient group. Deposits of C4d, mannose-binding lectin, C1q, IgM, and C5b-9 were scored in the glomeruli, peritubular capillaries, and arterioles. Notably, C4d deposits were present in 88.1% of TMA cases, and the various clinical conditions had distinct staining patterns within the various compartments of the renal vasculature. Classical pathway activation was observed in 90.5% of TMA cases. C5b-9 deposits were present in 78.6% of TMA cases and in 39.6% of controls (n=53), but the staining pattern differed between cases and controls. In conclusion, C4d is a common finding in TMA, regardless of the underlying clinical condition. Moreover, C5b-9 was present in >75% of the TMA samples, suggesting that terminal complement inhibitors may have a beneficial effect in these patients. C4d and C5b-9 should be investigated as possible diagnostic biomarkers in the clinical work-up of patients suspected of having complement-mediated TMA.
Asunto(s)
Complemento C4b/análisis , Enfermedades Renales/metabolismo , Glomérulos Renales/irrigación sanguínea , Glomérulos Renales/química , Fragmentos de Péptidos/análisis , Microangiopatías Trombóticas/metabolismo , Adolescente , Adulto , Anciano , Vasculitis Asociada a Anticuerpos Citoplasmáticos Antineutrófilos/metabolismo , Síndrome Antifosfolípido/metabolismo , Arteriolas/química , Biomarcadores/análisis , Capilares/química , Niño , Complemento C1q/análisis , Complejo de Ataque a Membrana del Sistema Complemento/análisis , Vía Clásica del Complemento , Femenino , Humanos , Inmunoglobulina M/análisis , Enfermedades Renales/complicaciones , Enfermedades Renales/patología , Glomérulos Renales/patología , Lupus Eritematoso Sistémico/metabolismo , Masculino , Lectina de Unión a Manosa/análisis , Persona de Mediana Edad , Microangiopatías Trombóticas/complicaciones , Microangiopatías Trombóticas/patología , Adulto JovenRESUMEN
Microcirculatory dysfunction may contribute to delayed cerebral ischemia after subarachnoid hemorrhage (SAH). Using a prechiasmatic injection model, this study investigated ultrastructural changes in microvessels in brain parenchyma to determine the nature of the microthromboemboli, the involvement of nitric oxide (NO) and P-selectin in their formation, and relationship to brain injury after SAH. Brains were examined by electron microscopy (EM) and immunohistochemistry. EM demonstrated that mice with SAH had significantly more arterioles filled with lesions consistent with microthrombi (in cortex, 20 ± 5 for SAH, 8 ± 4 saline-injected and 2.4 ± 0.2 for sham). SAH animals also had more constriction of arterioles. The concentration of NO was lower in mice with SAH (44 ± 9 for sham, 46 ± 20 for saline-injected and 24 ± 11 for SAH). The number of microthrombi correlated with the number of apoptotic neuronal cells (R(2)=0.80 in cortex). Cell membrane P-selectin increased in the endothelium of arterioles in mice with SAH (11.4 ± 0.7 for SAH, 6.8 ± 0.9 for sham and 6.1 ± 0.9 for saline-injected controls). This correlated with decreased NO in the brain. In conclusion, SAH causes microthrombosis and constriction of arterioles, which correlates with neuronal cell death. Increased P-selectin and decreased NO suggest a mechanism for microthrombosis and arteriolar constriction.
Asunto(s)
Trombosis Intracraneal/metabolismo , Trombosis Intracraneal/patología , Hemorragia Subaracnoidea/metabolismo , Hemorragia Subaracnoidea/patología , Animales , Apoptosis/fisiología , Arteriolas/química , Arteriolas/metabolismo , Arteriolas/ultraestructura , Modelos Animales de Enfermedad , Inmunohistoquímica , Etiquetado Corte-Fin in Situ , Masculino , Ratones , Microscopía Electrónica , Neuronas/patología , Óxido Nítrico/metabolismo , Selectina-P/metabolismo , Vasoconstricción/fisiologíaRESUMEN
BACKGROUND: Little is known about the relationship between splenic arteriolar hyaline and cause of death. The purpose of this retrospective study was to evaluate the clinicopathological significance of splenic arteriolar hyaline in autopsy cases and estimate the applicability of hyaline for diagnosing the cause and rapidity of death. METHODS: Archival data and histological slides from 82 cases were reviewed retrospectively. One section of each spleen was evaluated microscopically. The tinctorial pattern of splenic arteriolar hyaline was examined with Heidenhain's Azan trichrome stain, and the relationships between this pattern and age, cause of death, and rapidity of death were investigated. RESULTS: Fifty-four cases demonstrated hyaline change, with 3 different tinctorial patterns: red, blue, and a combination of red and blue. The 3 patterns coexisted in various proportions in each tissue section. Frequency of the blue pattern increased with age (P < 0.01) and was unrelated to cause of death. By contrast, the red pattern was unrelated to age and appeared with different frequency according to cause of death. The red pattern appeared with significantly higher frequency in the circulatory disease group and the drowning and asphyxia group (both P < 0.01). Moreover, the presence of the red pattern had high specificity for the detection of rapidly fatal cases. The combination of the 2 colors was excluded from clinicopathological analyses due to its admixed nature. CONCLUSIONS: Estimation of splenic arteriolar hyaline with Heidenhain's Azan trichrome stain is useful for assessment of the cause and rapidity of death. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1132441651796836.
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Arterioloesclerosis/complicaciones , Arterioloesclerosis/mortalidad , Muerte Súbita/etiología , Hialina/metabolismo , Bazo/irrigación sanguínea , Adolescente , Adulto , Factores de Edad , Anciano , Anciano de 80 o más Años , Arteriolas/química , Arteriolas/patología , Arterioloesclerosis/metabolismo , Arterioloesclerosis/patología , Autopsia , Causas de Muerte , Distribución de Chi-Cuadrado , Muerte Súbita/patología , Femenino , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Estudios Retrospectivos , Medición de Riesgo , Factores de Riesgo , Coloración y Etiquetado , Adulto JovenRESUMEN
Many of estrogen's effects on vascular reactivity are mediated through interaction with estrogen receptors (1, 2, 3). Although two sub-types exist (estrogen receptor -α and ß),estrogen receptor-α has been identified in both the smooth muscle and in endothelial cells of pial arterial segments using fluorescent staining combined with confocal laser scanning microscopy (4). Furthermore, ER-α is located in the nuclei and in the cytoplasm of rat basilar arteries (5). The receptors are abundant and fluoresce brightly, but clear visualization of discrete groups of receptors is difficult likely due to the numbers located in many cell layers of pial vessel segments. Additionally, many reports using immunohistochemical techniques paired with confocal microscopy poorly detail the requirements critical for reproduction of experiments (6). Our purpose for this article is to describe a simple technique to optimize the staining and visualization of ER-α using cross-sectional slices of pial arterioles obtain from female rat brains. We first perfuse rats with Evans blue dye to easily identify surface pial arteries which we isolate under a dissecting microscope. Use of a cryostat to slice 8 µm cross sections of the arteries allows us to obtain thin vessel sections so that different vessel planes are more clearly visualized. Cutting across the vessel rather than use of a small vessel segment has the advantage of easier viewing of the endothelial and smooth muscle layers. In addition, use of a digital immunofluorescent microscope with extended depth software produces clear images of ten to twelve different vessel planes and is less costly than use of a confocal laser scanning microscope.
Asunto(s)
Receptor alfa de Estrógeno/análisis , Microscopía Fluorescente/métodos , Piamadre/irrigación sanguínea , Animales , Arteriolas/química , Receptor alfa de Estrógeno/química , Femenino , RatasRESUMEN
Little is known about the impact of type 2 diabetes mellitus (DM) on coronary arteriole remodeling. The aim of this study was to determine the mechanisms that underlie coronary arteriole structural remodeling in type 2 diabetic (db/db) mice. Passive structural properties of septal coronary arterioles isolated from 12- to 16-week-old diabetic db/db and control mice were assessed by pressure myography. Coronary arterioles from 12-week-old db/db mice were structurally similar to age-matched controls. By 16 weeks of age, coronary wall thickness was increased in db/db arterioles (p < 0.01), while luminal diameter was reduced (control: 118 ± 5 µm; db/db: 102 ± 4 µm, p < 0.05), augmenting the wall-to-lumen ratio by 58% (control: 5.9 ± 0.6; db/db: 9.5 ± 0.4, p < 0.001). Inward hypertrophic remodeling was accompanied by a 56% decrease in incremental elastic modulus (p < 0.05, indicating decreased vessel coronary wall stiffness) and a ~30% reduction in coronary flow reserve (CFR) in diabetic mice. Interestingly, aortic pulse wave velocity and femoral artery incremental elastic modulus were increased (p < 0.05) in db/db mice, indicating macrovascular stiffness. Molecular tissue analysis revealed increased elastin-to-collagen ratio in diabetic coronaries when compared to control and a decrease in the same ratio in the diabetic aortas. These data show that coronary arterioles isolated from type 2 diabetic mice undergo inward hypertrophic remodeling associated with decreased stiffness and increased elastin-to-collagen ratio which results in a decreased CFR. This study suggests that coronary microvessels undergo a different pattern of remodeling from macrovessels in type 2 DM.
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Arteriolas/patología , Vasos Coronarios/patología , Diabetes Mellitus Tipo 2/patología , Elasticidad/fisiología , Animales , Arteriolas/química , Arteriolas/metabolismo , Colágeno Tipo I , Vasos Coronarios/química , Vasos Coronarios/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Elastina/análisis , Elastina/metabolismo , Masculino , Ratones , Ratones Mutantes , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
The heat and capsaicin receptor, TRPV1, is required for the detection of painful heat by primary afferent pain fibers (nociceptors), but the extent to which functional TRPV1 channels are expressed in the CNS is debated. Because previous evidence is based primarily on indirect physiological responses to capsaicin, here we genetically modified the Trpv1 locus to reveal, with excellent sensitivity and specificity, the distribution of TRPV1 in all neuronal and non-neuronal tissues. In contrast to reports of widespread and robust expression in the CNS, we find that neuronal TRPV1 is primarily restricted to nociceptors in primary sensory ganglia, with minimal expression in a few discrete brain regions, most notably in a contiguous band of cells within and adjacent to the caudal hypothalamus. We confirm hypothalamic expression in the mouse using several complementary approaches, including in situ hybridization, calcium imaging, and electrophysiological recordings. Additional in situ hybridization experiments in rat, monkey, and human brain demonstrate that the restricted expression of TRPV1 in the CNS is conserved across species. Outside of the CNS, we find TRPV1 expression in a subset of arteriolar smooth muscle cells within thermoregulatory tissues. Here, capsaicin increases calcium uptake and induces vasoconstriction, an effect that likely counteracts the vasodilation produced by activation of neuronal TRPV1.
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Arteriolas/metabolismo , Química Encefálica/genética , Regulación de la Expresión Génica , Genes Reporteros , Miocitos del Músculo Liso/metabolismo , Canales Catiónicos TRPV/biosíntesis , Animales , Arteriolas/química , Humanos , Hipotálamo/química , Hipotálamo/metabolismo , Macaca fascicularis , Masculino , Ratones , Ratones Transgénicos , Miocitos del Músculo Liso/química , Ratas , Ratas Sprague-Dawley , Canales Catiónicos TRPV/genética , Canales Catiónicos TRPV/fisiología , Vasoconstricción/genética , Vasodilatación/genéticaRESUMEN
Lipofuscin-like granules, first described by Biava and West in 1965, are a subcellular, quasi-physiologic finding mainly seen in the smooth muscle cells of renal arterioles, but also in juxtaglomerular cells and the lacis cells of human kidneys. They increase in number in subjects affected by arterial hypertension and diabetes. They do not correlate with a specific primary renal disease. Lipofuscin-like granules are not related to renin granules. The world literature on this subject is almost non-existent, and the awareness of this finding or its clinical significance among either pathologists or nephrologists is very poor. We incidentally observed these lipofuscin-like granules in 8 cases during the routine electron microscope examination of 440 renal biopsies, and report herein on their ultrastructural features. Six of these 8 patients were affected by arterial hypertension, one of whom was also concomitantly affected by diabetes mellitus. These lipofuscin-like granules appear as dense bodies with a lipid component, a coarsely granular matrix, and a crystalloid component which may appear in a band or dot pattern, according to the plane of sectioning. The pathologist has to be aware of these lipofuscin-like granules in order not to confuse them with the semicircularly organized (fingerprint) linear immune deposits associated with some specific glomerulopathies.
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Gránulos Citoplasmáticos/química , Gránulos Citoplasmáticos/ultraestructura , Aparato Yuxtaglomerular/química , Aparato Yuxtaglomerular/ultraestructura , Enfermedades Renales/diagnóstico , Lipofuscina/análisis , Adulto , Anciano , Arteriolas/química , Arteriolas/ultraestructura , Biopsia , Diabetes Mellitus , Femenino , Humanos , Hipertensión/complicaciones , Hallazgos Incidentales , Aparato Yuxtaglomerular/irrigación sanguínea , Enfermedades Renales/complicaciones , Enfermedades Renales/metabolismo , Enfermedades Renales/patología , Masculino , Microscopía Electrónica , Persona de Mediana Edad , Miocitos del Músculo Liso/química , Miocitos del Músculo Liso/ultraestructura , Estudios Retrospectivos , Adulto JovenRESUMEN
In arterioles, a locally initiated diameter change can propagate rapidly along the vessel length (arteriolar conducted response), thus contributing to arteriolar hemodynamic resistance. The response is underpinned by electrical coupling along the arteriolar endothelial layer. Connexins (Cx; constituents of gap junctions) are required for this coupling. This review addresses the effect of acute systemic inflammation (sepsis) on arteriolar conduction and interendothelial electrical coupling. Lipopolysaccharide (LPS; an initiating factor in sepsis) and polymicrobial sepsis (24 h model) attenuate conducted vasoconstriction in mice. In cultured microvascular endothelial cells harvested from rat and mouse skeletal muscle, LPS reduces both conducted hyperpolarization-depolarization along capillary-like structures and electrical coupling along confluent cell monolayers. LPS also tyrosine-phosphorylates Cx43 and serine-dephosphorylates Cx40. Since LPS-reduced coupling is Cx40- but not Cx43-dependent, only Cx40 dephosphorylation may be consequential. Nitric oxide (NO) overproduction is critical in advanced sepsis, since the removal of this overproduction prevents the attenuated conduction. Consistently, (i) exogenous NO in cultured cells reduces coupling in a Cx37-dependent manner, and (ii) the septic microvasculature in vivo shows no Cx40 phenotype. A complex role emerges for endothelial connexins in sepsis. Initially, LPS may reduce interendothelial coupling and arteriolar conduction by targeting Cx40, whereas NO overproduction in advanced sepsis reduces coupling and conduction by targeting Cx37 instead.
Asunto(s)
Arteritis/patología , Arteritis/fisiopatología , Conexinas/fisiología , Endotelio Vascular/patología , Endotelio Vascular/fisiopatología , Microvasos/patología , Microvasos/fisiopatología , Animales , Arteriolas/química , Arteriolas/patología , Arteriolas/fisiopatología , Arteritis/metabolismo , Endotelio Vascular/química , Humanos , Microvasos/química , Sepsis/metabolismo , Sepsis/patología , Sepsis/fisiopatología , Vasoconstricción/fisiologíaRESUMEN
AIMS: Angiotensin II (Ang II) is a strong renal vasoconstrictor and modulates the tubuloglomerular feedback (TGF). We hypothesized that Ang II at low concentrations enhances the vasoconstrictor effect of adenosine (Ado), the mediator of TGF. METHODS: Afferent arterioles of mice were isolated and perfused, and both isotonic contractions and cytosolic calcium transients were measured. RESULTS: Bolus application of Ang II (10(-12) and 10(-10) M) induced negligible vasoconstrictions, while Ang II at 10(-8) m reduced diameters by 35%. Ang II at 10(-12), 10(-10) and 10(-8) m clearly enhanced the arteriolar response to cumulative applications of Ado (10(-11) to 10(-4) M). Ado application increased the cytosolic calcium concentrations in the vascular smooth muscle, which were higher at 10(-5) M than at 10(-8) M. Ang II (10(-11) to 10(-6) M) also induced concentration-dependent calcium transients, which were attenuated by AT(1) receptor inhibition. Simultaneously applied Ang II (10(-10) M) additively enhanced the calcium transients induced by 10(-8) and 10(-5) M Ado. The transients were partly inhibited by AT(1) or A(1) receptor antagonists, but not significantly by A(2) receptor antagonists. CONCLUSION: A low dose of Ang II enhances Ado-induced constrictions, partly via AT(1) receptor-mediated calcium increase. Ado increases intracellular calcium by acting on A(1) but not A(2) receptors. The potentiating effect of Ang II on Ado-induced arteriolar vasoconstrictions may involve calcium sensitization of the contractile machinery, as Ang II only additively increased cytosolic calcium concentrations, while its effect on the arteriolar constriction was more than additive. The potentiating effect of Ang II might contribute to the resetting of TGF.
Asunto(s)
Adenosina/fisiología , Angiotensina II/fisiología , Calcio/química , Citosol/química , Riñón/irrigación sanguínea , Vasoconstricción/efectos de los fármacos , Adenosina/administración & dosificación , Antagonistas del Receptor de Adenosina A1 , Antagonistas del Receptor de Adenosina A2 , Angiotensina II/administración & dosificación , Bloqueadores del Receptor Tipo 1 de Angiotensina II/farmacología , Animales , Arteriolas/química , Arteriolas/efectos de los fármacos , Riñón/química , Masculino , Ratones , Ratones Endogámicos C57BL , Músculo Liso Vascular/química , Naftiridinas/farmacología , Purinas/farmacología , Receptor de Adenosina A1/metabolismo , Receptor de Angiotensina Tipo 1/metabolismo , Receptores de Adenosina A2/metabolismo , Triazinas/farmacología , Triazoles/farmacologíaRESUMEN
Adenosine can induce vasodilatation and vasoconstriction of the renal afferent arteriole of the mouse. We determined here its direct effect on efferent arterioles of mouse kidneys. Using isolated-perfused cortical efferent arterioles, we measured changes in luminal diameter in response to adenosine. Extraluminal application of adenosine and cyclohexyladenosine had no effect on the luminal diameter. When the vessels were constricted by the thromboxane mimetic U46619, application of adenosine and 5'-N-ethylcarboxamido-adenosine dilated the efferent arterioles in a dose-dependent manner. We also found that the adenosine-induced vasodilatation was inhibited by the A(2)-specific receptor blocker 3,7-dimethyl-1-propargylxanthine. In the presence of this inhibitor, adenosine failed to alter the basal vessel diameter of quiescent efferent arterioles. Using primer-specific polymerase chain reaction we found that the adenosine A(1), A(2a), A(2b), and A(3) receptors were expressed in microdissected mouse efferent arterioles. We conclude that adenosine dilates the efferent arteriole using the A(2) receptor subtype at concentrations compatible with activation of the A(2b) receptor.
Asunto(s)
Corteza Renal/irrigación sanguínea , Receptores de Adenosina A2/fisiología , Vasodilatación , Adenosina/análogos & derivados , Adenosina/farmacología , Animales , Arteriolas/química , Relación Dosis-Respuesta a Droga , Técnicas In Vitro , Ratones , Reacción en Cadena de la Polimerasa , Receptor de Adenosina A1/análisis , Receptor de Adenosina A1/genética , Receptor de Adenosina A2A/análisis , Receptor de Adenosina A2A/genética , Receptor de Adenosina A2B/análisis , Receptor de Adenosina A2B/genética , Receptor de Adenosina A3/análisis , Receptor de Adenosina A3/genética , Receptores de Adenosina A2/metabolismo , Vasodilatación/efectos de los fármacosRESUMEN
Cell adhesion molecule vascular endothelial cadherin (VE-cadherin) is the major component of endothelial adherence junctions, maintaining endothelial cell integrity. Studies dealing with constitutive VE-cadherin expression patterns in different pulmonary vessel types (arteries, arterioles, capillaries, venules, veins) or with the influence of physiological factors such as age or sex on VE-cadherin expression have not been published yet. Knowledge of constitutive resp. varying expression patterns not only fundamentally contribute to understanding the role of VE-cadherin in the pathogenesis of pulmonary diseases but also help to develop therapies based on immunotargeting. Hence, endothelial VE-cadherin expression was studied in regular lung tissue. Fifty-eight specimens of regular lung tissue (30 females, 28 males between 1 month and 75 years old) were immunohistochemically stained with an antibody against VE-cadherin. There was strong endothelial expression of VE-cadherin in arteries, arterioles, and capillaries but almost no expression in veins and venules. Neither age nor sex had any influence on the expression pattern or staining intensity. There is a vessel type-specific expression pattern for VE-cadherin in regular human lung tissue, which is not influenced by age or sex. Further studies will have to prove whether this is influenced by pathological conditions, e.g., ARDS.
Asunto(s)
Antígenos CD/análisis , Cadherinas/análisis , Endotelio Vascular/química , Pulmón/irrigación sanguínea , Arteria Pulmonar/química , Adolescente , Adulto , Factores de Edad , Anciano , Arteriolas/química , Capilares/química , Niño , Preescolar , Femenino , Humanos , Lactante , Masculino , Persona de Mediana Edad , Venas Pulmonares/química , Factores Sexuales , Vénulas/químicaRESUMEN
Although much physiology in resistance vessels has been attributed to the cytoplasmic connection between endothelial cells (ECs) and vascular smooth muscle cells (VSMCs), little is known of the protein expression between the two cell types. In an attempt to identify the proteins between ECs and VSMCs, mouse cremaster arterioles were stained with phalloidin-Alexa 594 and viewed on a confocal microscope that resolved "actin bridges" within the internal elastic lamina between ECs and VSMCs. To determine the incidence of protein, the pixel intensity from the antibodies on actin bridges were compared with the pixel intensity from antibodies within ECs or VSMCs. N-cadherin, desmin, connexin (Cx)40, and Cx43 and phosphorylated Cx43 at serine-368 were identified on actin bridges, but NG2, CD31, and Cx45 were not evident. Cx37 expression was more variable than the other connexins examined. Using this method on rat mesentery, we confirm the previously published predominance of Cx37 and Cx40 at the myoendothelial junction that was determined using electron microscopy. We conclude that this new method represents an important screening mechanism in which to rapidly test for protein expression between ECs and VSMCs and possibly a first-step in quantifying protein expression at the myoendothelial junction.
Asunto(s)
Actinas/análisis , Conexinas/análisis , Endotelio Vascular/química , Técnica del Anticuerpo Fluorescente/métodos , Uniones Intercelulares/química , Músculo Liso Vascular/química , Animales , Arteriolas/química , Colorantes Fluorescentes , Masculino , Ratones , Ratones Endogámicos C57BL , Microscopía Confocal , Compuestos Orgánicos , Faloidina , Fosforilación , RatasRESUMEN
Gap junctions are present in the juxtaglomerular apparatus enabling intercellular communication. Our study determined the location of different connexin subtypes within the juxtaglomerular apparatus of the rat, and the role of these subtypes in renal hemodynamics through the use of specific mimetic peptides. Immunohistochemical analysis showed connexins 37 and 40 expression in the endothelial and renin-secreting cells of the afferent arteriole, while connexin 40 was also found in extra- and intraglomerular mesangial cells. In contrast, connexin 43 was weakly expressed in endothelial cells of the afferent arteriole and within the glomerulus. Intra-renal infusion of the peptides (GAP) reported to block specific gap junctions ((Cx37,43)GAP27 or (Cx40)GAP27), elevated blood pressure, plasma renin activity, and angiotensin II levels, while decreasing renal plasma flow without a significant change in the glomerular filtration rate. Subsequent restoration of blood pressure reduced both renal plasma flow and glomerular filtration rate. In contrast, (Cx43)GAP26 reduced glomerular filtration rate without alterations in blood pressure, renal plasma flow, plasma renin activity, or angiotensin II levels. Hence, connexins 37 and 40 are expressed in the rat juxtaglomerular apparatus and these proteins control, in part, the renin-angiotensin system and renal autoregulation.