RESUMO
OBJECTIVE: Nonobstructive general angiography (NOGA) is a novel modality to detect and sample spontaneous ruptured aortic plaques (SRAPs). We aimed to establish novel methods to detect cholesterol crystals (CCs) in sampled SRAPs. METHODS: Blood specimens containing SRAPs were obtained from patients using NOGA. Blood was instantly frozen on a glass slide and subsequently thawed for quantitative analysis and spread onto a filter paper that was rinsed using distilled water. Qualitative analysis was performed for the rinsed water using polarized light microscopy, and the filter paper was embedded in paraffin for histologic analysis. RESULTS: The CCs were clearly observed after hemolysis using the instant freeze-thaw method. The filter paper rinse method indicated free CCs of varying shapes under polarized light microscopy without erythrocytes. On the filter paper, sampled SRAPs showed Lamé-like small particles. Histopathology revealed various atheromatous components. CONCLUSION: A set of novel methods for detecting CCs from sampled blood was established.
Assuntos
Placa Aterosclerótica , Aorta/química , Aorta/patologia , Colesterol , Humanos , Placa Aterosclerótica/diagnóstico , Placa Aterosclerótica/patologia , ÁguaRESUMO
SUMMARY: Formaldehyde (FA) is a toxic substance used frequently in the field of medicine as well as in many industrial areas. Especially people working in the field of anatomy, histology, and pathology are in high risk group because of the use of the FA. Studies showing the effects of FA on the cardiovascular system are few in number. The purpose of the present study was to investigate the effects of FA exposure, which we believe can cause oxidative stress, on the heart and aorta with various biochemical analyses. A total of 24 Wistar Albino rats were used in our study. We divided the rats into 3 groups as the Control Group (CG), the group exposed to low-dose FA (avg. 1 ppm) (DDG) Group, and the group exposed to high-dose FA (avg. 10 ppm) (YDG). At the end of the subchronic FA exposure, the blood samples, heart and aorta tissues of the rats were taken and subjected to biochemical analyses. As a result of the analyses, statistically significant differences were detected between CG (2.96?0.85 ng/mg), and HDG (2.08?0.77 ng/mg) in aortic tissues in TXNIP analysis (p<0.05). In heart tissues, significant differences were detected between CG (0.73?0.27 ng/mg) and LDG (1.13?0.22 ng/mg) (p<0.05). Statistically significant differences were also detected between CG (1.98?0.31 mM/ml) and YDG (2.43?0.31 mM/ml) in serum MDA analyses (p<0.05). It was shown that subchronic application of FA to LDG rats through inhalation had no effects on apoptosis markers in heart tissues. More studies are required to show FA toxicity and the mechanism of action of pathology on the cardiovascular system. We believe that our study will contribute to clarifying the roles of mild and subchronic exposure of FA in heart and aortic tissues in terms of oxidative stress risk.
RESUMEN: El formaldehído es una sustancia tóxica que se utiliza con frecuencia en el campo de la medicina, así como en muchas áreas industriales. Especialmente las personas que trabajan en el area de la anatomía, y patología se encuentran en el grupo de alto riesgo debido al uso de esta sustancia. Pocos son los estudios que muestran los efectos del formaldehído en el sistema cardiovascular. El propósito del presente estudio fue investigar a través de análisis bioquímicos, los efectos de la exposición a formaldehído, que podría causar estrés oxidativo, en el corazón y la aorta. Se utilizaron un total de 24 ratas Albinas Wistar. Dividimos a las ratas en 3 grupos: grupo control (GC), grupo expuesto a dosis bajas de AG (promedio 1 ppm) (DDG) y grupo expuesto a dosis altas de AG (promedio 10 ppm) (YDG). Al término de la exposición a FA subcrónica, se tomaron muestras de sangre, tejido cardíaco y aorta de las ratas y se sometieron a análisis bioquímicos. Como resultado de los análisis, se detec- taron diferencias estadísticamente significativas entre GC (2,96 ? 0,85 ng / mg) y HDG (2,08 ? 0,77 ng / mg) en los tejidos aórticos en el análisis TXNIP (p <0,05). En los tejidos cardíacos se detectaron diferencias significativas entre GC (0,73 ? 0,27 ng / mg) y LDG (1,13 ? 0,22 ng / mg) (p <0,05). También se detectaron diferencias estadísticamente significativas entre CG (1,98 ? 0,31 mM / ml) y YDG (2,43 ? 0,31 mM / ml) en los análisis de MDA en suero (p <0,05). Se demostró que la aplicación subcrónica de formaldehído a ratas LDG a través de la inhalación no tuvo efectos sobre los marcadores de apoptosis en los tejidos del corazón. Se requieren más estudios para demostrar la toxicidad de los AG y el mecanismo de acción de la patología en el sistema cardiovascular. Creemos que nuestro estudio contribuirá a aclarar las funciones de la exposición leve y subcrónica de formaldehído en los tejidos cardíacos y aórticos en términos de riesgo al estrés oxidativo.
Assuntos
Animais , Ratos , Aorta/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Formaldeído/farmacologia , Coração/efeitos dos fármacos , Aorta/química , Tiorredoxinas/análise , Fenômenos Bioquímicos , Inalação , Ratos Wistar , Peroxidase/análise , Formaldeído/administração & dosagem , Hidroxiprolina/análise , Miocárdio/químicaRESUMO
Atherosclerosis is a multifactorial disease, for which the etiology is so complex that we are currently unable to prevent it and effectively lower the statistics on mortality from cardiovascular diseases. Parallel to modern analyses in molecular biology and biochemistry, we want to carry out analyses at the level of micro- and macroelements in order to discover the interdependencies between elements during atherogenesis. In this work, we used the Inductively Coupled Plasma Optical Emission Spectrometer (ICP-OES) to determine the content of calcium, magnesium, iron, copper, chromium, zinc, manganese, cadmium, lead, and zinc in the aorta sections of people who died a sudden death. We also estimated the content of metalloenzymes MMP-9, NOS-3, and SOD-2 using the immunohistochemical method. It was observed that with the age of the patient, the calcium content of the artery increased, while the content of copper and iron decreased. Very high correlations (correlation coefficient above 0.8) were observed for pairs of parameters in women: Mn-Ca, Fe-Cu, and Ca-Cd, and in men: Mn-Zn. The degree of atherosclerosis negatively correlated with magnesium and with cadmium. Chromium inhibited absorption of essential trace elements such as Cu and Fe due to its content being above the quantification threshold only if Cu and Fe were lower. Moreover, we discussed how to design research for the future in order to learn more about the pathomechanism of atherosclerosis and the effect of taking dietary supplements on the prevalence of cardiovascular diseases.
Assuntos
Aorta/química , Aorta/metabolismo , Aterosclerose/metabolismo , Espectrofotometria Atômica/métodos , Oligoelementos/análise , Oligoelementos/metabolismo , Idoso , Idoso de 80 Anos ou mais , Aorta/patologia , Aterosclerose/patologia , Cádmio/análise , Cádmio/metabolismo , Cálcio/análise , Cálcio/metabolismo , Cobre/análise , Cobre/metabolismo , Feminino , Humanos , Ferro/análise , Ferro/metabolismo , Magnésio/análise , Magnésio/metabolismo , Masculino , Manganês/análise , Manganês/metabolismo , Pessoa de Meia-IdadeRESUMO
Medial degeneration is a common histopathological finding in aortopathy and is considered a mechanism for dilatation. We investigated if medial degeneration is specific for sporadic thoracic aortic aneurysms versus nondilated aortas. Specimens were graded by pathologists, blinded to the clinical diagnosis, according to consensus histopathological criteria. The extent of medial degeneration by qualitative (semi-quantitative) assessment was not specific for aneurysmal compared to nondilated aortas. In contrast, blinded quantitative assessment of elastin amount and medial cell number distinguished aortic aneurysms and referent specimens, albeit with marked overlap in results. Specifically, the medial fraction of elastin decreased from dilution rather than loss of protein as cross-sectional amount was maintained while the cross-sectional number, though not density, of smooth muscle cells increased in proportion to expansion of the media. Furthermore, elastic lamellae did not thin and interlamellar distance did not diminish as expected for lumen dilatation, implying a net gain of lamellar elastin and intralamellar cells or extracellular matrix during aneurysmal wall remodeling. These findings support the concepts that: (1) medial degeneration need not induce aortic aneurysms, (2) adaptive responses to altered mechanical stresses increase medial tissue, and (3) greater turnover, not loss, of mural cells and extracellular matrix associates with aortic dilatation.
Assuntos
Aorta/anatomia & histologia , Aneurisma da Aorta Torácica/patologia , Túnica Média/ultraestrutura , Adaptação Fisiológica , Adulto , Idoso , Aorta/química , Doença da Válvula Aórtica Bicúspide/patologia , Contagem de Células , Comorbidade , Elastina/análise , Matriz Extracelular/ultraestrutura , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Miócitos de Músculo Liso/ultraestrutura , Método Simples-Cego , Coloração e Rotulagem , Remodelação VascularRESUMO
When decomposition of a recovered body is fairly advanced, identification based on common morphologic features is often impossible. In these cases, short tandem repeat (STR) marker genotyping has established itself as a convenient and reliable alternative. However, at very progressed stages of decomposition, postmortem tissue putrefaction processes can decrease DNA yields considerably. Hence, not all types of tissue are equally suitable for successful STR marker-based postmortem identification. Bone or dental material is often analysed in corpses with advanced decompositional changes. However, processing of these materials is very elaborate and time and resource consuming. We have therefore focused on the suitableness of various types of soft tissue swabs, where DNA extraction is easier and faster. By sampling 28 bodies at various stages of decomposition, we evaluated the suitability of different tissues for genotyping at varying degrees of physical decay. This was achieved by a systematic classification of the sampled bodies by morphological scoring and subsequent analysis of multiple tissue swabs of the aortic wall, urinary bladder wall, brain, liver, oral mucosa and skeletal muscle. In summary, we found variable degrees of suitability of different types of soft tissue swabs for DNA-based identification. Swabs of the aortic wall, the urinary bladder wall and brain tissue yielded the best results - in descending order - even at advanced levels of decay.
Assuntos
Restos Mortais/química , DNA/isolamento & purificação , Antropologia Forense/métodos , Aorta/química , Química Encefálica , Degradação Necrótica do DNA , Impressões Digitais de DNA/métodos , Feminino , Humanos , Fígado/química , Masculino , Repetições de Microssatélites , Mucosa Bucal/química , Músculo Esquelético/química , Mudanças Depois da Morte , Bexiga Urinária/químicaRESUMO
The ingestion of excessive free fatty acid could induce lipotoxicity in tissues and then lead to the initiation of many metabolism diseases. In this work, the protective effect of apigenin on palmitate-induced lipotoxicity in human aortic endothelial cells (HAEC) was investigated. Compared with 150 µM palmitate treatment alone, pretreatment with 10 µM apigenin for 6 h significantly increased the cell viability from 71.55 ± 3.62 to 91.06 ± 4.30% and improved mitochondrial membrane potential to the normal level (101.62 ± 11.72% of control). In addition, the production of nitric oxide was markedly elevated by apigenin cotreatment from 7.10 ± 3.95 to 94.20 ± 21.86%. The data-independent acquisition-based proteomic approach was used to study the protective mechanism, and the results revealed that 242 proteins were differently expressed in cells treated with palmitate and 93 proteins were reversed after apigenin supplementation. Apigenin realized its protective function mainly via regulating pathways such as IL-17, TNF, Fox O, cell adhesion, and endoplasmic reticulum protein processing. Collectively, these data demonstrated that apigenin supplement may serve as an alternative nutritional intervention to protect HAEC against lipotoxicity.
Assuntos
Aorta/citologia , Apigenina/farmacologia , Células Endoteliais/efeitos dos fármacos , Palmitatos/toxicidade , Substâncias Protetoras/farmacologia , Animais , Aorta/química , Aorta/efeitos dos fármacos , Aorta/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Células Endoteliais/química , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Humanos , Óxido Nítrico/metabolismo , Proteínas/química , Proteínas/genética , Proteínas/metabolismo , ProteômicaRESUMO
Catheterization is a common medical operation to diagnose and treat cardiovascular diseases. The blood vessel lumen is coated with endothelial glycocalyx layer (EGL), which is important for the permeability and diffusion through the blood vessels wall, blood hemodynamics and mechanotransduction. However EGL's role in catheter-blood vessel friction is not explored. We use a porcine aorta to mimic the blood vessel and a catheter loop was made to rub in reciprocating sliding mode against it to understand the role of catheter loop curvature, stiffness, normal load, sliding speed and EGL on the friction properties. Trypsin treatment was used to cause a degradation of the EGL. Decrease in catheter loop stiffness and EGL degradation were the strongest factors which dramatically increased the coefficient of friction (COF) and frictional energy dissipation at the aorta-catheter interface. Increasing sliding speed caused an increase but increase in normal load first caused a decrease and then an increase in the COF and frictional energy. These results provide the basic data for safety of operation and damage control during catheterization in patients with degraded EGL.
Assuntos
Aorta/química , Endotélio Vascular/química , Glicocálix/química , Mecanotransdução Celular/fisiologia , Animais , Aorta/efeitos dos fármacos , Fenômenos Biomecânicos , Cateterismo/efeitos adversos , Endotélio Vascular/efeitos dos fármacos , Fricção , Glicocálix/efeitos dos fármacos , Hemodinâmica/fisiologia , Humanos , Permeabilidade , Suínos , Técnicas de Cultura de Tecidos , Tripsina/farmacologia , Dispositivos de Acesso Vascular/efeitos adversosRESUMO
BACKGROUND: Our previous research revealed that trypsin is abundantly expressed in atherosclerotic plaques and its distribution overlaps with that of matrix metalloproteinase-9 (MMP-9). This study was performed to explore the possible roles of trypsin in vulnerable atherosclerotic plaque formation. METHODS AND RESULTS: Twenty-four rabbits were randomly assigned to a normal (control) group, an atherosclerosis (experimental) group and a trypsin inhibitor (aprotinin) group. In the 13th feeding week, the aprotinin group was treated with 5 mg/kg/day aprotinin via ear vein for 4 weeks. At the end of the 16th week, coronary arterial and aortic expression of trypsin, proteinase-activated receptor-2 (PAR-2), activated MMP-9, and pro-inflammatory cytokines were significantly greater in the experimental group than in the control group. Aprotinin decreased trypsin expression and activation in plaques, blocked PAR-2 and MMP-9 activation, and decreased cytokine expression; it also increased fibrous cap thickness, decreased the intima-media thickness and intimal/medial ratio, thus significantly ameliorating plaque vulnerability. Upregulated trypsin, MMP-9 and PAR-2 were also found in coronary intimal atherosclerotic plaques of patients undergoing coronary artery bypass grafting. CONCLUSIONS: Ectopic trypsin was significantly upregulated in atherosclerotic plaques, which increased pro-inflammatory cytokine levels by activating PAR-2 and promoted plaque instability by activating proMMP-9, thereby promoting atherosclerosis and plaque vulnerability. In addition, the high trypsin expression in human coronary intimal atherosclerotic plaques suggests that targeting trypsin may be a new strategy for acute coronary syndrome prevention.
Assuntos
Aterosclerose/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Placa Aterosclerótica/química , Tripsina/metabolismo , Animais , Aorta/química , Aprotinina/administração & dosagem , Aprotinina/metabolismo , Espessura Intima-Media Carotídea , Citocinas/metabolismo , Regulação da Expressão Gênica , Humanos , Masculino , Coelhos , Receptor PAR-2/metabolismo , Tripsina/genética , Inibidores da Tripsina/administração & dosagem , Inibidores da Tripsina/metabolismoRESUMO
BACKGROUND: The prevalence of hypertension in young women is lower than that in age-matched men while the prevalence of hypertension in women is significantly increased after the age of 50 (menopause) and is greater than that in men. It is already known that sphingosine-1-phosphate (S1P) and ceramide regulate vascular tone with opposing effects. This study aimed to explore the effects of ovariectomy and estrogen supplementation on the ceramide/S1P rheostat of the aorta in rats, and to explore a potential mechanism for perimenopausal hypertension and a brand-new target for menopausal hormone therapy to protect vessels. METHODS: In total, 30 female adult SD rats were randomly divided into three groups: The sham operation group (SHAM), ovariectomy group (OVX) and ovariectomy plus estrogen group (OVX + E). After 4 weeks of treatment, the blood pressure (BP) of the rats was monitored by a noninvasive system; the sphingolipid content (e.g., ceramide and S1P) was detected by liquid chromatography-mass spectrometry (LC-MS); the expression of the key enzymes involved in ceramide anabolism and catabolism was measured by real-time fluorescence quantitative polymerase chain reaction (qPCR); and the expression of key enzymes and proteins in the sphingosine kinase 1/2 (SphK1/2)-S1P-S1P receptor 1/2/3 (S1P1/2/3) signaling pathway was detected by qPCR and western blotting. RESULTS: In the OVX group compared with the SHAM group, the systolic BP (SBP), diastolic BP (DBP) and pulse pressure (PP) increased significantly, especially the SBP and PP (P < 0.001). For aortic ceramide metabolism, the mRNA level of key enzymes involved in anabolism and catabolism decreased in parallel 2-3 times, while the contents of total ceramide and certain long-chain subtypes increased significantly (P < 0.05). As for the S1P signaling pathway, SphK1/2, the key enzymes involved in S1P synthesis, decreased significantly, and the content of S1P decreased accordingly (P < 0.01). The S1P receptors showed various trends: S1P1 was significantly down-regulated, S1P2 was significantly up-regulated, and S1P3 showed no significant difference. No significant difference existed between the SHAM and OVX + E groups for most of the above parameters (P > 0.05). CONCLUSIONS: Ovariectomy resulted in the imbalance of the aortic ceramide/S1P rheostat in rats, which may be a potential mechanism underlying the increase in SBP and PP among perimenopausal women. Besides, the ceramide/S1P rheostat may be a novel mechanism by which estrogen protects vessels.
Assuntos
Aorta/metabolismo , Ceramidas/metabolismo , Estrogênios/uso terapêutico , Hipertensão/prevenção & controle , Lisofosfolipídeos/metabolismo , Pós-Menopausa/efeitos dos fármacos , Esfingosina/análogos & derivados , Animais , Aorta/química , Ceramidas/análise , Estrogênios/farmacologia , Feminino , Hipertensão/tratamento farmacológico , Hipertensão/etiologia , Hipertensão/metabolismo , Lisofosfolipídeos/análise , Modelos Animais , Ovariectomia , Ratos , Ratos Sprague-Dawley , Esfingosina/análise , Esfingosina/metabolismoRESUMO
A major limitation in extrusion-based bioprinting is the lack of direct process control, which limits the accuracy and design complexity of printed constructs. The lack of direct process control results in a number of defects that can influence the functional and mechanical outcomes of the fabricated structures. The machine axes motion cannot be reliably used to predict material placement, and precise fabrication requires additional sensing of the material extrusion. We present an iteration-to-iteration process monitoring system that enables direct process control in the material deposition reference frame. To fabricate parts with low dimensional errors, we integrate a non-contact laser displacement scanner into the printing platform. After fabrication of the initial print using the as-designed reference trajectory, the laser scanner moves across the part to measure the material placement. A custom image processing algorithm compares the laser scanner data to the as-designed reference trajectory to generate an error vector. To compensate for the measured error, the algorithm modifies the axes reference trajectory for the second print iteration. We implement the in situ process monitoring and error compensation technique on an experimental platform to evaluate system performance and demonstrate improvement in spatial material placement.
Assuntos
Bioimpressão/métodos , Algoritmos , Aorta/química , Aorta/citologia , Bioimpressão/instrumentação , Bioimpressão/normas , Impressão Tridimensional/instrumentação , Alicerces Teciduais/químicaRESUMO
OBJECTIVE: Marfan syndrome (MFS) is caused by mutations in FBN1 (fibrillin-1), an extracellular matrix (ECM) component, which is modified post-translationally by glycosylation. This study aimed to characterize the glycoproteome of the aortic ECM from patients with MFS and relate it to aortopathy. Approach and Results: ECM extracts of aneurysmal ascending aortic tissue from patients with and without MFS were enriched for glycopeptides. Direct N-glycopeptide analysis by mass spectrometry identified 141 glycoforms from 47 glycosites within 35 glycoproteins in the human aortic ECM. Notably, MFAP4 (microfibril-associated glycoprotein 4) showed increased and more diverse N-glycosylation in patients with MFS compared with control patients. MFAP4 mRNA levels were markedly higher in MFS aortic tissue. MFAP4 protein levels were also increased at the predilection (convexity) site for ascending aorta aneurysm in bicuspid aortic valve patients, preceding aortic dilatation. In human aortic smooth muscle cells, MFAP4 mRNA expression was induced by TGF (transforming growth factor)-ß1 whereas siRNA knockdown of MFAP4 decreased FBN1 but increased elastin expression. These ECM changes were accompanied by differential gene expression and protein abundance of proteases from ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family and their proteoglycan substrates, respectively. Finally, high plasma MFAP4 concentrations in patients with MFS were associated with a lower thoracic descending aorta distensibility and greater incidence of type B aortic dissection during 68 months follow-up. CONCLUSIONS: Our glycoproteomics analysis revealed that MFAP4 glycosylation is enhanced, as well as its expression during the advanced, aneurysmal stages of MFS compared with control aneurysms from patients without MFS.
Assuntos
Aorta/química , Matriz Extracelular/química , Glicopeptídeos/análise , Síndrome de Marfan/metabolismo , Proteômica/métodos , Aneurisma da Aorta Torácica/metabolismo , Proteínas de Transporte/sangue , Proteínas de Transporte/genética , Proteínas de Transporte/fisiologia , Proteínas da Matriz Extracelular/sangue , Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/fisiologia , Fibrilina-1/genética , Glicoproteínas/sangue , Glicoproteínas/genética , Glicoproteínas/fisiologia , Glicosilação , Humanos , Miócitos de Músculo Liso/metabolismo , Remodelação VascularRESUMO
Decellularized porcine aorta (PA) is a promising biomaterial for vascular substitutes. However, decellularized PAs suffer from mechanical weakness and have less pores, which limit cellular ingrowth into the grafts and hinder the remodeling. In this study, PAs were decellularized by vacuum-freeze-thawing cycles and 0.3% of sodium dodecyl sulfate (SDS) buffer (VLS). Results showed that the application of vacuum-freeze-thawing significantly improved the decellularization efficiency of SDS while effectively preserved the mechanical function of PA tissues, decreased residual SDS, and minimized cytotoxicity. Furthermore, scanning electron microscopy (SEM) examination demonstrated that VLS generated interconnected pores with uniform distribution. In vivo subcutaneous implantation assay further demonstrated that VLS implants had less calcification and adverse inflammatory response. Moreover, VLS treatment markedly enhanced ingrowth of myofibroblasts and endothelial cells, and thereby promoted synthesis of extracellular matrix and vascularization. These results suggest that the application of vacuum-freeze-thawing into the decellularization process may produce a promising vascular graft candidate for tissue engineering application.
Assuntos
Aorta/transplante , Aorta/ultraestrutura , Engenharia Tecidual , Alicerces Teciduais , Animais , Aorta/química , Aorta/citologia , Fenômenos Biomecânicos , Proliferação de Células , Células Cultivadas , Células Endoteliais da Veia Umbilical Humana , Humanos , Porosidade , Ratos , Suínos , Engenharia Tecidual/métodos , Alicerces Teciduais/químicaRESUMO
High-quality matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) of lipids in biological tissue relies on the fabrication of a homogeneous matrix coating featuring best possible analyte integration. This communication addresses a matrix vapor deposition/recrystallization process for the application of 1,5-diaminonaphthalene (1,5-DAN) onto slices of human aortic tissue. The matrix coating is compatible with both positive- as well as negative-ion-mode MALDI MSI facilitating a significantly enhanced detection of lipid-related signals in different cell layers of blood vessel walls. Graphical abstract.
Assuntos
Aorta/química , Lipídeos/análise , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , 2-Naftilamina/análogos & derivados , 2-Naftilamina/química , Aorta/ultraestrutura , Temperatura Baixa , Humanos , Coloração e Rotulagem , VácuoRESUMO
Prostaglandin E2 (PGE2) plays an important role in vascular homeostasis. Its receptor, E-prostanoid receptor 4 (EP4) is essential for physiological remodeling of the ductus arteriosus (DA). However, the role of EP4 in pathological vascular remodeling remains largely unknown. We found that chronic angiotensin II (AngII) infusion of mice with vascular smooth muscle cell (VSMC)-specific EP4 gene knockout (VSMC-EP4-/-) frequently developed aortic dissection (AD) with severe elastic fiber degradation and VSMC dedifferentiation. AngII-infused VSMC-EP4-/- mice also displayed more profound vascular inflammation with increased monocyte chemoattractant protein-1 (MCP-1) expression, macrophage infiltration, matrix metalloproteinase-2 and -9 (MMP2/9) levels, NADPH oxidase 1 (NOX1) activity, and reactive oxygen species production. In addition, VSMC-EP4-/- mice exhibited higher blood pressure under basal and AngII-infused conditions. Ex vivo and in vitro studies further revealed that VSMC-specific EP4 gene deficiency significantly increased AngII-elicited vasoconstriction of the mesenteric artery, likely by stimulating intracellular calcium release in VSMCs. Furthermore, EP4 gene ablation and EP4 blockade in cultured VSMCs were associated with a significant increase in MCP-1 and NOX1 expression and a marked reduction in α-SM actin (α-SMA), SM22α, and SM differentiation marker genes myosin heavy chain (SMMHC) levels and serum response factor (SRF) transcriptional activity. To summarize, the present study demonstrates that VSMC EP4 is critical for vascular homeostasis, and its dysfunction exacerbates AngII-induced pathological vascular remodeling. EP4 may therefore represent a potential therapeutic target for the treatment of AD.
Assuntos
Angiotensina II/metabolismo , Dissecção Aórtica/metabolismo , Pressão Sanguínea/fisiologia , Inflamação/metabolismo , Receptores de Prostaglandina E Subtipo EP4 , Animais , Aorta/química , Aorta/metabolismo , Aneurisma Aórtico/metabolismo , Dinoprostona/metabolismo , Feminino , Hipertensão/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Músculo Liso Vascular/metabolismo , Receptores de Prostaglandina E Subtipo EP4/genética , Receptores de Prostaglandina E Subtipo EP4/metabolismo , Remodelação Vascular/genéticaRESUMO
BACKGROUND: A bicuspid aortic valve (BAV) is the most common congenital cardiac malformation and is associated with ascending aortic dilation in 60%-80% of patients. In this study, we aimed to address the role of hemodynamic influences on the development of aortopathy in BAV patients. PATIENT AND METHODS: BAV (n=36) and tricuspid aortic valve (TAV) patients (n=17) undergoing aortic valve replacement underwent preoperative flow magnetic resonance imaging (MRI) assessment to detect the area of maximal flow-induced stress in the proximal aorta. Based on these MRI data, paired ascending aortic wall samples [i.e., area of maximal jet impact (jet sample) and the opposite aortic wall (nonjet sample)] were collected during surgery. To study and describe the effects of jet stream on the complete vascular wall, a pathology score was developed based on the recently published aortic consensus paper statement on surgical pathology of the aorta using routine histologic stainings (resorcin fuchsin, hematoxylin-eosin, and Movat) and immunohistochemistry (alpha smooth muscle actin, smooth muscle 22 alpha, platelet endothelial cell adhesion molecule). RESULTS: Comparing the jet and nonjet samples in both BAV and TAV, regions of maximal jet impact did not show any difference in the pathology score in the adventitia and the middle and outer media. In the jet samples, the inner media however showed loss of actin expression in both BAV (P<.0001) and the TAV (P=.0074), and the intimal thickness was significantly enlarged in both patient groups (BAV P=.0005, TAV P=.0041), which was not accompanied by loss of elastic lamellae or vascular smooth muscle cell nuclei. CONCLUSIONS: In our study population, we could not demonstrate a potential distinct role for hemodynamics in the development of aortopathy in BAV patients even if corrected for aortic diameter, raphe position, or whether the valve is stenotic or regurgitant. The intimal layer and inner media however showed alterations in all jet specimens.
Assuntos
Aorta/fisiopatologia , Aneurisma Aórtico/fisiopatologia , Valva Aórtica/anormalidades , Doenças das Valvas Cardíacas/fisiopatologia , Hemodinâmica , Actinas/análise , Idoso , Aorta/química , Aorta/diagnóstico por imagem , Aorta/patologia , Aneurisma Aórtico/diagnóstico por imagem , Aneurisma Aórtico/etiologia , Aneurisma Aórtico/metabolismo , Valva Aórtica/diagnóstico por imagem , Valva Aórtica/fisiopatologia , Insuficiência da Valva Aórtica/etiologia , Insuficiência da Valva Aórtica/fisiopatologia , Estenose da Valva Aórtica/etiologia , Estenose da Valva Aórtica/fisiopatologia , Doença da Válvula Aórtica Bicúspide , Dilatação Patológica , Feminino , Doenças das Valvas Cardíacas/complicações , Doenças das Valvas Cardíacas/diagnóstico por imagem , Humanos , Imuno-Histoquímica , Imageamento por Ressonância Magnética , Masculino , Proteínas dos Microfilamentos/análise , Pessoa de Meia-Idade , Proteínas Musculares/análise , Molécula-1 de Adesão Celular Endotelial a Plaquetas/análiseRESUMO
Trace elements are crucial for vital enzymatic reactions in all metabolic processes. Zinc (Zn) acts as a co-factor for many enzymes. Copper (Cu) and iron (Fe) have pro-atherogenic effects resulting in atherosclerosis. Aorta exposing high pulsatile pressure is sensitive for atherosclerosis because of its fast metabolism and poor nutrition by diffusion from vasa vasorum. We aimed to determine the relationship between serum Zn, Cu and Fe levels with aortic and left internal mammary artery (LIMA) tissues in 33 atherosclerotic individuals who inevitably underwent coronary artery by-pass graft (CBAG) surgery that is an end-point treatment for atherosclerosis. Trace elements in serum and tissues were measured using inductively coupled plasma-optical emission spectrophotometer. Pre-operative (Pre-op) serum Fe levels were statistically 46% higher than post-operative (Post-op) values (p = 0.009). Aortic Fe level was 49.8% higher than LIMA Fe (p = 0.0001). Our study points out the tendency of aortic tissue to atherosclerosis via pro- atherogenic effect of Fe. LIMA, being a potential graft for CBAG, is resistant to atherosclerosis with its intimal specialty of graft patency. In conclusion, serum Zn, Cu and Fe levels in atherosclerotic CBAG patients might be monitored to reveal minor alterations pre-operatively and post-operatively for ameliorating the treatment and life quality.
Assuntos
Aorta/química , Artérias/química , Aterosclerose/cirurgia , Cobre/análise , Ponte de Artéria Coronária , Vasos Coronários/química , Ferro/análise , Zinco/análise , Aterosclerose/patologia , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Synchrotron radiation (SR) based X-ray imaging is an attractive method for analyzing biomedical structure. However, despite its many advantages, there are few gold standards in image processing methods, especially in segmentation. Image segmentation is an essential step in medical imaging for image analysis and three-dimensional reconstruction. Although there are many algorithms for image segmentation, a decisive method does not exist in SR X-ray imagery, because of a lack of data. This study focused on finding a suitable algorithm for image segmentation in high-resolution medical imaging. In this study, we used following four algorithms to segment blood vessel of mouse; interactive graph cuts algorithm, which segments an image using fast min-cut/max-flow algorithm to solve global solution, binary partition tree algorithm, which uses an interactive method creating tree nodes to segment an image by using splitting and merging an image, seeded region growing algorithm, which performs segmentation by connecting similar pixel value, simple interactive object extraction, which generates color signature for segmentation based on color model of an image.
Assuntos
Vasos Sanguíneos/química , Algoritmos , Animais , Aorta/anatomia & histologia , Aorta/química , Vasos Sanguíneos/anatomia & histologia , Cor , Aumento da Imagem , Processamento de Imagem Assistida por Computador , Masculino , Camundongos , Ratos , Ratos Wistar , Tomografia Computadorizada por Raios XRESUMO
Thyroid hormone (TH) regulates gene transcription by binding to TH receptors (TRs). TRs regulate the genes of lipid metabolism and the renin-angiotensin system (RAS). We examined the effect of TRα deletion in ApoE-/- mice (DKO mice) on the following: (i) the expression of genes controlling cholesterol metabolism and tissue (t)RAS in the liver and aorta and (ii) the expression of these genes and the regulation of cholesterol content in cultured vascular smooth muscle cells (VSMCs). TRα deletion in ApoE-/- mice led to the repression of genes involved in the synthesis and influx of cholesterol in the liver. However, TRα deletion in the arterial wall suppressed the expression of genes involved in the esterification and excretion of cholesterol and enhanced the expression of angiotensinogen (AGT). The VSMCs of the ApoE-/- and DKO mice increased their cholesterol content during cholesterol loading, but failed to increase the expression of ATP-binding cassette transporter A1 (ABCA1). T3 addition partially corrected these abnormalities in the cells of the ApoE-/- mice but not those of the DKO mice. In conclusion, TRα deletion in ApoE-/- mice slightly increases the expression of tRAS in the aorta and aggravates the dysregulation of cholesterol content in the VSMCs.
Assuntos
Apolipoproteínas E/deficiência , Colesterol/metabolismo , Músculo Liso Vascular/metabolismo , Sistema Renina-Angiotensina/fisiologia , Receptores alfa dos Hormônios Tireóideos/deficiência , Transportador 1 de Cassete de Ligação de ATP/genética , Animais , Aorta/química , Apolipoproteínas E/genética , Apolipoproteínas E/fisiologia , Aterosclerose/diagnóstico por imagem , Células Cultivadas , Colesterol/administração & dosagem , Colesterol/genética , Expressão Gênica , Hibridização Genética , Fígado/química , Masculino , Camundongos , Camundongos Knockout , Músculo Liso Vascular/química , Músculo Liso Vascular/citologia , RNA Mensageiro , Receptores alfa dos Hormônios Tireóideos/genética , Receptores alfa dos Hormônios Tireóideos/fisiologia , Tri-Iodotironina/farmacologia , UltrassonografiaRESUMO
BACKGOUND: The inability to detect premature atherosclerosis significantly hinders implementation of personalized therapy to prevent coronary heart disease. A comprehensive understanding of arterial protein networks and how they change in early atherosclerosis could identify new biomarkers for disease detection and improved therapeutic targets. METHODS: Here we describe the human arterial proteome and proteomic features strongly associated with early atherosclerosis based on mass spectrometry analysis of coronary artery and aortic specimens from 100 autopsied young adults (200 arterial specimens). Convex analysis of mixtures, differential dependent network modeling, and bioinformatic analyses defined the composition, network rewiring, and likely regulatory features of the protein networks associated with early atherosclerosis and how they vary across 2 anatomic distributions. RESULTS: The data document significant differences in mitochondrial protein abundance between coronary and aortic samples (coronary>>aortic), and between atherosclerotic and normal tissues (atherosclerotic<Assuntos
Aorta/química
, Doenças da Aorta/metabolismo
, Aterosclerose/metabolismo
, Doença da Artéria Coronariana/metabolismo
, Vasos Coronários/química
, Proteínas/análise
, Proteômica/métodos
, Espectrometria de Massas em Tandem
, Adolescente
, Adulto
, Aorta/patologia
, Doenças da Aorta/patologia
, Aterosclerose/patologia
, Autopsia
, Biomarcadores/análise
, Doença da Artéria Coronariana/patologia
, Vasos Coronários/patologia
, Feminino
, Humanos
, Masculino
, Pessoa de Meia-Idade
, Placa Aterosclerótica
, Mapas de Interação de Proteínas
, Adulto Jovem
RESUMO
We aimed to identify the potential genes related to blood pressure regulation and screen target genes for high blood pressure (BPH) and low blood pressure (BPL) treatment. The GSE19817 microarray dataset, which included the aorta, liver, heart, and kidney samples from BPH, BPL, and normotensive mice, was downloaded from the Gene Expression Omnibus. Principal component analysis (PCA) was performed based on the entire expression profile. Differentially expressed genes (DEGs) were screened, followed by pathway enrichment analysis. Finally, gene regulatory networks were constructed based on BPH-related and BPL-related DEGs in the aorta, liver, heart, and kidney samples. As a result, DEGs were screened within their respective tissues due to high heterogeneity of different tissues. Totally, 2,726 BPH-related DEGs and 2,472 BPL-related DEGs were screened, which were mainly enriched in pathways such as immune response. The topology data of gene regulatory networks constructed by DEGs in the heart, kidney, and liver were similar than that in aorta. Finally, among BPH-related DEGs, Sept6 and Pigx were found in the top 10 differentially regulated DEGs by comparing the BPH-related DEGs of the aorta with the DEGs of the other 3 tissues in the regulatory network. Although among the top 10 differentially regulated BPL-related DEGs, no common differentially regulated DEGs were found, Wif1, Urb2, and Gtf2ird1 were found among the top ten DEGs in the three tissues other than the kidney tissue. Sept6 and Pigx might participate in the pathogenesis of BPH, whereas Gtf2ird1, Urb2, and Wif1 might be critical target genes for BPL treatment.