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1.
Int J Mol Sci ; 22(13)2021 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-34209949

RESUMO

Fibrinolysis is a complex enzymatic process aimed at dissolving blood clots to prevent vascular occlusions. The fibrinolytic system is composed of a number of cofactors that, by regulating fibrin degradation, maintain the hemostatic balance. A dysregulation of fibrinolysis is associated with various pathological processes that result, depending on the type of abnormality, in prothrombotic or hemorrhagic states. This narrative review is focused on the congenital and acquired disorders of primary fibrinolysis in both adults and children characterized by a hyperfibrinolytic state with a bleeding phenotype.


Assuntos
Transtornos da Coagulação Sanguínea/metabolismo , Fibrinólise , Hemorragia/metabolismo , Adulto , Transtornos da Coagulação Sanguínea/etiologia , Criança , Redes Reguladoras de Genes , Hemorragia/etiologia , Humanos
2.
Int J Mol Sci ; 22(13)2021 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-34203338

RESUMO

Diffuse alveolar hemorrhage (DAH) in systemic lupus erythematosus (SLE) is associated with significant mortality, requiring a thorough understanding of its complex mechanisms to develop novel therapeutics for disease control. Activated p53-dependent apoptosis with dysregulated long non-coding RNA (lncRNA) expression is involved in the SLE pathogenesis and correlated with clinical activity. We examined the expression of apoptosis-related p53-dependent lncRNA, including H19, HOTAIR and lincRNA-p21 in SLE-associated DAH patients. Increased lincRNA-p21 levels were detected in circulating mononuclear cells, mainly in CD4+ and CD14+ cells. Higher expression of p53, lincRNA-p21 and cell apoptosis was identified in lung tissues. Lentivirus-based short hairpin RNA (shRNA)-transduced stable transfectants were created for examining the targeting efficacy in lncRNA. Under pristane stimulation, alveolar epithelial cells had increased p53, lincRNA-p21 and downstream Bax levels with elevated apoptotic ratios. After pristane injection, C57/BL6 mice developed DAH with increased pulmonary expression of p53, lincRNA-p21 and cell apoptosis. Intra-pulmonary delivery of shRNA targeting lincRNA-p21 reduced hemorrhage frequencies and improved anemia status through decreasing Bax expression and cell apoptosis. Our findings demonstrate increased p53-dependent lncRNA expression with accelerated cell apoptosis in the lungs of SLE-associated DAH patients, and show the therapeutic potential of targeting intra-pulmonary lncRNA expression in a pristane-induced model of DAH.


Assuntos
RNA Longo não Codificante/genética , Proteína Supressora de Tumor p53/metabolismo , Animais , Apoptose/fisiologia , Modelos Animais de Doenças , Feminino , Hemorragia/genética , Hemorragia/metabolismo , Humanos , Pulmão/metabolismo , Pulmão/microbiologia , Pneumopatias/genética , Pneumopatias/metabolismo , Lúpus Eritematoso Sistêmico/genética , Lúpus Eritematoso Sistêmico/metabolismo , Masculino , Alvéolos Pulmonares/metabolismo , Alvéolos Pulmonares/microbiologia , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Proteína Supressora de Tumor p53/genética
3.
Int J Mol Sci ; 22(13)2021 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-34206377

RESUMO

Infiltration of red blood cells into atheromatous plaques and oxidation of hemoglobin (Hb) and lipoproteins are implicated in the pathogenesis of atherosclerosis. α1-microglobulin (A1M) is a radical-scavenging and heme-binding protein. In this work, we examined the origin and role of A1M in human atherosclerotic lesions. Using immunohistochemistry, we observed a significant A1M immunoreactivity in atheromas and hemorrhaged plaques of carotid arteries in smooth muscle cells (SMCs) and macrophages. The most prominent expression was detected in macrophages of organized hemorrhage. To reveal a possible inducer of A1M expression in ruptured lesions, we exposed aortic endothelial cells (ECs), SMCs and macrophages to heme, Oxy- and FerrylHb. Both heme and FerrylHb, but not OxyHb, upregulated A1M mRNA expression in all cell types. Importantly, only FerrylHb induced A1M protein secretion in aortic ECs, SMCs and macrophages. To assess the possible function of A1M in ruptured lesions, we analyzed Hb oxidation and heme-catalyzed lipid peroxidation in the presence of A1M. We showed that recombinant A1M markedly inhibited Hb oxidation and heme-driven oxidative modification of low-density lipoproteins as well plaque lipids derived from atheromas. These results demonstrate the presence of A1M in atherosclerotic plaques and suggest its induction by heme and FerrylHb in the resident cells.


Assuntos
alfa-Globulinas/metabolismo , Aterosclerose/etiologia , Aterosclerose/metabolismo , Heme/metabolismo , Hemoglobinas/metabolismo , Peroxidação de Lipídeos , Oxirredução , Aterosclerose/patologia , Biomarcadores , Doenças das Artérias Carótidas/etiologia , Doenças das Artérias Carótidas/metabolismo , Doenças das Artérias Carótidas/patologia , Células Cultivadas , Progressão da Doença , Suscetibilidade a Doenças , Hemorragia/metabolismo , Hemorragia/patologia , Humanos , Imuno-Histoquímica , Miócitos de Músculo Liso/metabolismo , Placa Aterosclerótica/etiologia , Placa Aterosclerótica/metabolismo , Placa Aterosclerótica/patologia
4.
Int J Biol Macromol ; 182: 179-186, 2021 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-33838185

RESUMO

Peroxisome proliferator-activated receptor α (PPARα) play a key role in the regulation of metabolic homeostasis, inflammation, cellular growth, and differentiation. To further explore the potential role of PPARα in the energy homeostasis of fatty liver hemorrhagic syndrome (FLHS), we reported the prokaryotic expression and purification of chicken PPARα subunit protein, and successfully prepared a polyclonal antibody against PPARα recombinant protein. The 987 bp PPARα subunit genes were cloned into the pEASY-T3 clone vector. Then the plasmid PCR products encoding 329 amino acids were ligated to pEASY-Blunt E2 vector and transformed into BL21 to induce expression. The recombinant PPARα subunit protein, containing His-tag, was purified by affinity column chromatography using Ni-NTA affinity column. Rabbit antiserum was generated by using the concentration of recombinant PPARα subunit protein as the antigen. The results of western blotting showed that the antiserum can specifically recognize chicken endogenous PPARα protein. Immunohistochemistry and immunofluorescence showed that the PPARα mainly existed in the nucleus of hepatocytes, renal epithelial cells and hypothalamic endocrine nerve cells. More importantly, western blotting and real-time quantitative PCR indicated that FLHS significantly decreased the expression of PPARα.


Assuntos
Anticorpos/imunologia , Fígado Gorduroso/veterinária , Hemorragia/veterinária , PPAR alfa/metabolismo , Doenças das Aves Domésticas/metabolismo , Animais , Reações Antígeno-Anticorpo , Western Blotting/métodos , Células Cultivadas , Galinhas , Fígado Gorduroso/metabolismo , Feminino , Hemorragia/metabolismo , Hepatócitos/metabolismo , Hipotálamo/metabolismo , Imuno-Histoquímica/métodos , Rim/metabolismo , PPAR alfa/genética , PPAR alfa/imunologia , Síndrome
5.
Int J Hematol ; 114(2): 172-178, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33907978

RESUMO

Lipopolysaccharide (LPS) and tissue factor (TF) have frequently been used to induce disseminated intravascular coagulation (DIC) in experimental animal models. We have previously reported that the pathophysiology of DIC differs according to the inducing agents. However, inflammatory status and bleeding symptoms have not been fully compared between rat models of the two forms of DIC. We attempted to evaluate detailed characteristic features of LPS- and TF-induced DIC models, especially in regard to inflammatory status and bleeding symptoms, in addition to selected hemostatic parameters and pathologic findings in the kidneys. The degree of hemostatic activation in both types of experimental DIC was identical, based on the results of thrombin-antithrombin complex levels. Markedly elevated tumor necrosis factor, interleukin-6, and high-mobility group box-1 concentrations were observed with severe organ dysfunction and marked fibrin deposition in the kidney on administration of LPS, whereas markedly elevated D-dimer concentration and bleeding symptoms were observed with TF administration. Pathophysiology such as fibrinolytic activity, organ dysfunction, inflammation status, and bleeding symptom differed markedly between LPS- and TF-induced DIC models in rats. We, therefore, recommend that these disease models be assessed carefully as distinct entities to determine the implications of their experimental and clinical use.


Assuntos
Suscetibilidade a Doenças , Coagulação Intravascular Disseminada/complicações , Coagulação Intravascular Disseminada/etiologia , Hemorragia/etiologia , Hemorragia/metabolismo , Lipopolissacarídeos/efeitos adversos , Tromboplastina/efeitos adversos , Animais , Biomarcadores , Coagulação Sanguínea , Testes de Coagulação Sanguínea , Modelos Animais de Doenças , Coagulação Intravascular Disseminada/sangue , Coagulação Intravascular Disseminada/diagnóstico , Hemorragia/diagnóstico , Humanos , Masculino , Prognóstico , Ratos
6.
J Surg Res ; 261: 43-50, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33412508

RESUMO

BACKGROUND: Monitoring of decrease in fibrinogen levels with surgical blood loss is crucial for timely transfusion of fresh frozen plasma (FFP) to avoid coagulopathic bleeding. Here, we validated a simulation model to predict hemorrhagic reductions in fibrinogen levels during major noncardiac surgery. METHODS: We retrospectively performed exponential regression analysis of intraoperative blood loss and fibrinogen levels to develop a simulation model in the initial 50 patients and applied the model to another 59 patients to compare the measured and simulated fibrinogen levels. We examined the relationship between FFP transfusion and the measured fibrinogen levels or blood loss. The fibrinogen trigger level of FFP transfusion was below 130 mg/dL, although the decision of a perioperative blood transfusion was at the discretion of the anesthesiologists and surgeons. RESULTS: Application of the simulation model based on the initial 50 patients to another 59 patients showed no difference between the measured and estimated fibrinogen levels (189 ± 61 versus 186 ± 62, P = 0.60, mean difference: -2.28, limits of agreement: -69.42 to 64.84). The estimated fibrinogen level (mg/dL) = preoperative fibrinogen × exp (-1.90 × [blood loss/estimated circulation volume]), in which the estimated circulation volume = (70 [mL/kg] × body weight [kg]). FFP transfusion was significantly related to the measured fibrinogen level (cutoff: 145; 95% confidence intervals: 124-168; P = 0.0003) but not blood loss (P = 0.12). CONCLUSIONS: Fibrinogen level simulation predicted a hemorrhagic fibrinogen decline, thereby guiding FFP transfusion during active surgical bleeding. Further studies on the usefulness of fibrinogen level simulation are warranted.


Assuntos
Perda Sanguínea Cirúrgica , Transfusão de Sangue , Fibrinogênio/metabolismo , Hemorragia/metabolismo , Modelos Biológicos , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos
7.
Indian J Pathol Microbiol ; 64(1): 117-122, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33433420

RESUMO

Context: Hemophilia A is classified as mild, moderate, and severe based on Factor VIII levels (FVIII). Clot-based assays only detect initiation of thrombin generation, hence FVIII levels may not accurately predict the bleeding risk in all hemophilia patients. The entire process of thrombin generation as measured by global hemostasis tests like activated partial thromboplastin time clot waveform analysis (APTT CWA) and thrombin generation test (TGT) may reflect the actual bleeding phenotype. Aims: To assess the utility of TGT and CWA as a screening tool to identify bleeders and to evaluate the bleeding phenotype in Hemophilia A. Settings and Design: Prospective, observational study of 147 consecutive patients referred for coagulation workup. Subjects and Methods: Bleeding assessment tool was used to identify bleeders. Patients were classified as severe and nonsevere bleeders based on clinical criteria. TGT was performed by calibrated automated thrombogram, CWA by photo-optical coagulometer and factor levels by one stage clot-based assays. Statistical Analysis Used: The Kruskal-Wallis test with post-hoc analysis was done to examine the difference in CWA/TGT parameters amongst hemophilia classified by FVIII levels. Receiver operating characteristic (ROC) analysis was performed to estimate the diagnostic accuracy of CWA and TGT in discriminating between clinically severe vs nonsevere bleeders. Results: Using ROC derived cut-offs of min1, min2 and peak height of thrombin (PH), the sensitivity (min1:91.67%, min2:91.67%, PH: 97.22%, FVIII: 86.11%) and specificity (min1:100%, min2:100%, PH: 90.91%, FVIII: 90.91%) of CWA/TGT was superior to FVIII to distinguish between clinically severe vs nonsevere bleeders. Phenotypic heterogeneity of bleeding severity was identified in our study population. Clinical severity correlated with CWA/TGT parameters instead of FVIII levels. Conclusions: CWA and TGT are more effective tools than conventional factor assays to identify clinically severe bleeders and tailor prophylaxis as per bleeding phenotype.


Assuntos
Hemorragia/metabolismo , Tempo de Tromboplastina Parcial/normas , Fenótipo , Trombina/análise , Trombose , Testes de Coagulação Sanguínea/normas , Hemofilia A/diagnóstico , Hemorragia/classificação , Humanos , Tempo de Tromboplastina Parcial/métodos , Estudos Prospectivos , Curva ROC , Trombina/metabolismo
8.
J Photochem Photobiol B ; 214: 112087, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33234463

RESUMO

Bothrops leucurus is the major causative agent of snakebites in Brazil's Northeast. The systemic effects of its venom are effectively neutralized by antivenom therapy, preventing bitten patients' death. However, antivenom fails in neutralizing local effects that include intense pain, edema, bleeding, and myonecrosis. Such effects can lead to irreversible sequels, representing a clinically relevant issue for which there is no current effective treatment. Herein, the effects of photobiomodulation therapy (PBMT) were tested in the local actions induced by B. leucurus venom (BLV) in mice (n = 123 animals in 20 experimental groups). A continuous emission AlGaAs semiconductor diode laser was used in two wavelengths (660 or 780 nm). Mechanical nociceptive thresholds were assessed with the electronic von Frey apparatus. Local edema was determined by measuring the increase in paw thickness. Hemorrhage was quantified by digital measurement of the bleeding area. Myotoxicity was evaluated by serum creatine kinase (CK) activity and histopathological analysis. PBMT promoted anti-hypernociception in BLV-injected mice; irradiation with the 660 nm laser resulted in faster effect onset than the 780 nm laser. Both laser protocols reduced paw edema formation, whether irradiation was performed immediately or half an hour after venom injection. BLV-induced hemorrhage was not altered by PBMT. Laser irradiation delayed, but did not prevent myotoxicity caused by BLV, as shown by a late increase in CK activity and histopathological alterations. PBMT was effective in the control of some of the major local effects of BLV refractory to antivenom. It is a potential complementary therapy that could be used in bothropic envenoming, minimizing the morbidity of these snakebite accidents.


Assuntos
Antivenenos/química , Edema/radioterapia , Terapia com Luz de Baixa Intensidade/métodos , Mordeduras de Serpentes/radioterapia , Animais , Antivenenos/metabolismo , Bothrops , Creatina Quinase/sangue , Creatina Quinase/metabolismo , Edema/induzido quimicamente , Hemorragia/metabolismo , Hemorragia/radioterapia , Humanos , Lasers Semicondutores , Masculino , Camundongos , Músculo Esquelético/efeitos da radiação , Necrose/radioterapia
9.
Life Sci ; 266: 118913, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-33333050

RESUMO

AIM: Cyclophosphamide (CYP) chemotherapy induces bladder toxicity and hemorrhagic cystitis in cancer patients constituting a current clinical concern. Oxidative inflammatory cascades have been implicated as the mechanism contributing to CYP bladder urotoxicity. We thus assayed to explore whether zinc (Zn) supplementation could mitigate CYP-induced urotoxicity and evaluate the possible underlying mechanism in rats. MAIN METHOD: Rats were orally administered Zn (100 mg/kg b.w./day) for 10 days against urotoxicity induced by single injection of CYP (150 mg/kg b.w., ip) on day 7. KEY FINDINGS: CYP significantly depressed bladder activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and reduced glutathione (GSH) levels, whereas malondialdehyde level was increased prominently. In addition, CYP induced marked increases in the levels of tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), interleukin-6 (IL-6), and nitric oxide (NO) confirmed by histological alterations. CYP prominently increased bladder inducible nitric oxide synthase (iNOS) activity, nuclear factor-kappa B (NF-ĸB) and expression of caspase-3 protein. Zinc supplementation considerably abrogated the bladder urotoxicity by restoring redox balance, proinflammatory and apoptotic cascades and alleviated histopathological changes. SIGNIFICANCE: This is the first to reveal zinc potential to prevent CYP-induced urotoxic hemorrhagic cystitis via restoring redox balance and enhancing anti-inflammatory and antiapoptotic mechanisms in rat bladder.


Assuntos
Ciclofosfamida/toxicidade , Cistite/prevenção & controle , Suplementos Nutricionais , Regulação da Expressão Gênica/efeitos dos fármacos , Hemorragia/prevenção & controle , Zinco/farmacologia , Animais , Antineoplásicos Alquilantes/toxicidade , Caspase 3/química , Caspase 3/genética , Caspase 3/metabolismo , Cistite/induzido quimicamente , Cistite/metabolismo , Cistite/patologia , Hemorragia/induzido quimicamente , Hemorragia/metabolismo , Hemorragia/patologia , Masculino , NF-kappa B/antagonistas & inibidores , NF-kappa B/genética , NF-kappa B/metabolismo , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Ratos , Ratos Wistar
10.
PLoS One ; 15(12): e0243207, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33264359

RESUMO

BACKGROUND: Intramyocardial hemorrhage following reperfusion is strongly associated with major adverse cardiovascular events in myocardial infarction (MI) patients; yet the mechanisms contributing to these outcomes are not well understood. Large animal models have been used to investigate intramyocardial hemorrhage, but they are exorbitantly expensive and difficult to use for mechanistic studies. In contrast, rat models are widely used to investigate mechanistic aspects of cardiovascular physiology, but a rat model that consistently recapitulates the characteristics of an hemorrhagic MI does not exist. To bridge this gap, we investigated the physiological conditions of MI that would create intramyocardial hemorrhage in rats so that a reliable model of hemorrhagic MI would become available for basic research. METHODS & RESULTS: Sprague-Dawley rats underwent either a 90-minute (90-min) ischemia and then reperfusion (I/R) (n = 22) or 30-minute (30-min) I/R (n = 18) of the left anterior descending coronary artery. Sham rats (n = 12) were used as controls. 90-min I/R consistently yielded hemorrhagic MI, while 30-min I/R consistently yielded non-hemorrhagic MI. Twenty-four hours post-reperfusion, ex-vivo late-gadolinium-enhancement (LGE) and T2* cardiac MRI performed on excised hearts from 90-min I/R rats revealed colocalization of iron deposits within the scarred tissue; however, in 30-min I/R rats scar was evident on LGE but no evidence of iron was found on T2* CMR. Histological studies verified tissue damage (H&E) detected on LGE and the presence of iron (Perl's stain) observed on T2*-CMR. At week 4 post-reperfusion, gene and protein expression of proinflammatory markers (TNF-α, IL-1ß and MMP-9) were increased in the 90-min I/R group when compared to 30-min I/R groups. Further, transmission electron microscopy performed on 90-min I/R myocardium that were positive for iron on T2* CMR and Perl's stain showed accumulation of granular iron particles within the phagosomes. CONCLUSION: Ischemic time prior to reperfusion is a critical factor in determining whether a MI is hemorrhagic or non-hemorrhagic in rats. Specifically, a period of 90-min of ischemia prior to reperfusion can produce rat models of hemorrhagic MI, while 30-minutes of ischemia prior to reperfusion can ensure that the MIs are non-hemorrhagic. Hemorrhagic MIs in rats result in marked increase in iron deposition, proinflammatory burden and adverse left-ventricular remodeling compared to rats with non-hemorrhagic MIs.


Assuntos
Biomarcadores/metabolismo , Hemorragia/diagnóstico por imagem , Infarto do Miocárdio/diagnóstico por imagem , Traumatismo por Reperfusão Miocárdica/complicações , Animais , Modelos Animais de Doenças , Gadolínio/administração & dosagem , Hemorragia/etiologia , Hemorragia/genética , Hemorragia/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Imagem Cinética por Ressonância Magnética , Masculino , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Infarto do Miocárdio/etiologia , Infarto do Miocárdio/genética , Infarto do Miocárdio/metabolismo , Traumatismo por Reperfusão Miocárdica/diagnóstico por imagem , Ratos , Ratos Sprague-Dawley , Fatores de Tempo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Regulação para Cima
11.
Toxicol Lett ; 335: 91-97, 2020 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-33157172

RESUMO

Nemopilema nomurai venom (NnV) is severely toxic to many organisms. However, the mechanism of its poisoning has not been properly understood yet. The present work demonstrates that zebrafish (Danio rerio) is an alternative vertebrate model for studying NnV jellyfish venom for the first time. In this model, NnV appears to cause severe hemorrhage and inflammation in cardiopulmonary regions of zebrafish. NnV also altered the swimming behavior of zebrafish accompanied by a significant downregulation of acetylcholinesterase (AChE) activity in brain tissues. Histopathological changes observed for various organs of D. rerio caused by NnV corresponded to an increase in lactate dehydrogenase (LDH) activity in tissues. NnV also significantly altered glutathione S-transferase (GST) activity in cardiopulmonary and brain tissues of D. rerio. SDS-PAGE revealed many protein bands of NnV of various sizes after silver staining. Taken together, these results indicate that Danio rerio can be a useful alternative animal model for jellyfish venom toxicology studies. Findings of the present study also suggest that Danio rerio could be used to develop an effective treatment strategy and discover the mechanism of action of jellyfish venom envenomation.


Assuntos
Venenos de Cnidários/toxicidade , Modelos Animais de Doenças , Hemorragia/induzido quimicamente , Síndromes Neurotóxicas/etiologia , Cifozoários/química , Peixe-Zebra , Animais , Biomarcadores/metabolismo , Peso Corporal/efeitos dos fármacos , Venenos de Cnidários/isolamento & purificação , Relação Dose-Resposta a Droga , Coração/efeitos dos fármacos , Hemorragia/metabolismo , Hemorragia/patologia , Miocárdio/patologia , Síndromes Neurotóxicas/metabolismo , Síndromes Neurotóxicas/patologia , Tamanho do Órgão/efeitos dos fármacos , Especificidade de Órgãos , Sistema Respiratório/efeitos dos fármacos , Sistema Respiratório/patologia
12.
Int Heart J ; 61(6): 1129-1134, 2020 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-33191348

RESUMO

Patients with impaired kidney function have a high frequency of intraplaque hemorrhage (IPH) in their coronary arteries. Levels of cyclophilin A (CyPA), an indirect matrix metalloproteinase inducer, are increased in deceased patients who had impaired kidney function. In this study, we have examined the relationship between IPH and CyPA.We examined 47 samples of coronary plaque from 27 cadavers with coronary stenosis. These sections, all with > 50% coronary stenosis, were stained with an antibody against CyPA and the expression of CyPA was semi-quantified. Cadavers and plaques were classified into one of two groups depending on the presence or absence of IPH. IPH was defined as the presence of red blood cells stained with hematoxylin and eosin (HE) indicative of overt acute hemorrhage.In an individual analysis, estimation of glomerular filtration rate (eGFR) in the IPH group was significantly lower than that in the non-IPH group (P = 0.002). In a histological analysis, the percentage of stained area of CyPA in the IPH group was significantly higher than that in the non-IPH group (P < 0.0001).IPH was associated with a significantly higher expression of CyPA in this study. In addition, patients with IPH in their coronary arteries had significantly impaired kidney function.


Assuntos
Estenose Coronária/metabolismo , Ciclofilina A/metabolismo , Hemorragia/metabolismo , Placa Aterosclerótica/metabolismo , Insuficiência Renal/metabolismo , Idoso , Idoso de 80 Anos ou mais , Cadáver , Estenose Coronária/complicações , Estenose Coronária/patologia , Vasos Coronários/metabolismo , Vasos Coronários/patologia , Feminino , Taxa de Filtração Glomerular , Hemorragia/complicações , Hemorragia/patologia , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Placa Aterosclerótica/complicações , Placa Aterosclerótica/patologia , Insuficiência Renal/complicações
13.
FASEB J ; 34(11): 15252-15268, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32959379

RESUMO

Sphingolipids have been implicated in mammalian placental development and function, but their regulation in the placenta remains unclear. Herein we report that alkaline ceramidase 2 (ACER2) plays a key role in sustaining the integrity of the placental vasculature by regulating the homeostasis of sphingolipids in mice. The mouse alkaline ceramidase 2 gene (Acer2) is highly expressed in the placenta between embryonic day (E) 9.5 and E12.5. Acer2 deficiency in both the mother and fetus decreases the placental levels of sphingolipids, including sphingoid bases (sphingosine and dihydrosphingosine) and sphingoid base-1-phosphates (sphingosine-1-phosphate and dihydrosphingosine-1-phosphate) and results in the in utero death of ≈50% of embryos at E12.5 whereas Acer2 deficiency in either the mother or fetus has no such effects. Acer2 deficiency causes hemorrhages from the maternal vasculature in the junctional and/or labyrinthine zones in E12.5 placentas. Moreover, hemorrhagic but not non-hemorrhagic Acer2-deficient placentas exhibit an expansion of parietal trophoblast giant cells with a concomitant decrease in the area of the fetal blood vessel network in the labyrinthine zone, suggesting that Acer2 deficiency results in embryonic lethality due to the atrophy of the fetal blood vessel network in the placenta. Taken together, these results suggest that ACER2 sustains the integrity of the placental vasculature by controlling the homeostasis of sphingolipids in mice.


Assuntos
Ceramidase Alcalina/fisiologia , Hemorragia/patologia , Lisofosfolipídeos/metabolismo , Placenta/patologia , Esfingolipídeos/metabolismo , Esfingosina/análogos & derivados , Doenças Vasculares/patologia , Animais , Feminino , Hemorragia/etiologia , Hemorragia/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Placenta/metabolismo , Gravidez , Esfingosina/metabolismo , Doenças Vasculares/etiologia , Doenças Vasculares/metabolismo
14.
Int J Mol Sci ; 21(18)2020 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-32942605

RESUMO

As the main particulate component of the circulating blood, RBCs play major roles in physiological hemodynamics and impact all arterial wall pathologies. RBCs are the main determinant of blood viscosity, defining the frictional forces exerted by the blood on the arterial wall. This function is used in phylogeny and ontogeny of the cardiovascular (CV) system, allowing the acquisition of vasomotricity adapted to local metabolic demands, and systemic arterial pressure after birth. In pathology, RBCs collide with the arterial wall, inducing both local retention of their membranous lipids and local hemolysis, releasing heme-Fe++ with a high toxicity for arterial cells: endothelial and smooth muscle cells (SMCs) cardiomyocytes, neurons, etc. Specifically, overloading of cells by Fe++ promotes cell death. This local hemolysis is an event associated with early and advanced stages of human atherosclerosis. Similarly, the permanent renewal of mural RBC clotting is the major support of oxidation in abdominal aortic aneurysm. In parallel, calcifications promote intramural hemorrhages, and hemorrhages promote an osteoblastic phenotypic shift of arterial wall cells. Different plasma or tissue systems are able, at least in part, to limit this injury by acting at the different levels of this system.


Assuntos
Artérias/metabolismo , Artérias/patologia , Aterosclerose/metabolismo , Aterosclerose/patologia , Eritrócitos/metabolismo , Eritrócitos/fisiologia , Hemoglobinas/metabolismo , Animais , Hemólise/fisiologia , Hemorragia/metabolismo , Hemorragia/patologia , Humanos , Lipídeos de Membrana/metabolismo , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia
15.
Biomed Chromatogr ; 34(12): e4963, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32789887

RESUMO

Raw Moutan Cortex (RMC) and Processed Moutan Cortex (PMC) have a long history of use in China and other Asian countries. In this study, a rapid and accurate ultra-high-pressure liquid chromatography coupled with diode array detector (UHPLC-DAD) method was developed and validated for the simultaneous determination of nine absorbed compounds of RMC/PMC. After extraction by protein precipitation with methanol from plasma, the analytes were separated on an Acquity UPLC® BEH Shield RP18 column (2.1 × 100 mm, 1.7 µm, Waters, USA). Acetonitrile (A) and 0.1% (v/v) formic acid in water (B) were selected as the mobile phase to perform gradient elution. The linearity of nine analytes was >0.9915. The intra- and inter-assay precision (RSD) values were within 11.18%, and accuracy ranged from 91.32 to 101.29%. Suitable stability, matrix effect and extraction recoveries were also obtained. The validated method was applied to compare the pharmacokinetics of RMC and PMC in Blood-Heat and Hemorrhage Syndrome Model and normal rats. The results revealed that processing and the pathological state could influence the pharmacokinetic characteristics of compounds in RMC/PMC. The study willbe useful for further studies on pharmacokinetics and clinical application of raw and processed Moutan Cortex.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas , Hemorragia/metabolismo , Medicina Tradicional Chinesa , Paeonia , Espectrometria de Massas em Tandem/métodos , Animais , Benzoatos/sangue , Benzoatos/farmacocinética , Modelos Animais de Doenças , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/análise , Medicamentos de Ervas Chinesas/farmacocinética , Glucosídeos/sangue , Glucosídeos/farmacocinética , Limite de Detecção , Modelos Lineares , Masculino , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Terpenos/sangue , Terpenos/farmacocinética
16.
Radiat Res ; 194(2): 162-172, 2020 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-32845987

RESUMO

Thrombocytopenia (TCP) may cause severe and life-threatening bleeding. While this may be prevented by platelet transfusions, transfusions are associated with potential complications, do not always work (platelet refractory) and are not always available. There is an urgent need for a synthetic alternative. We evaluated the ability of fibrinogen-coated nanospheres (FCNs) to prevent TCP-related bleeding. FCNs are made of human albumin polymerized into a 100-nm sphere and coated with fibrinogen. We hypothesized that FCNs would bind to platelets through fibrinogen-GPIIb/IIIa interactions, contributing to hemostasis in the setting of TCP. We used two murine models to test these effects: in the first model, BALB/c mice received 7.25 Gy total-body irradiation (TBI); in the second model, lower dose TBI (7.0 Gy) was combined with an anti-platelet antibody (anti-CD41) to induce severe TCP. Deaths in both models were due to gastrointestinal or intracranial bleeding. Addition of antiplatelet antibody to 7.0 Gy TBI significantly worsened TCP and increased mortality compared to 7.0 Gy TBI alone. FCNs significantly improved survival compared to saline control in both models, suggesting it ameliorated TCP-related bleeding. Additionally, in a saphenous vein bleeding model of antibody-induced TCP, FCNs shortened bleeding times. There were no clinical or histological findings of thrombosis or laboratory findings of disseminated intravascular coagulation after FCN treatment. In support of safety, fluorescence microscopy suggests that FCNs bind to platelets only upon platelet activation with collagen, limiting activity to areas of endothelial damage. To our knowledge, this is the first biosynthetic agent to demonstrate a survival advantage in TCP-related bleeding.


Assuntos
Albuminas/química , Fibrinogênio/química , Fibrinogênio/farmacologia , Hemorragia/complicações , Hemorragia/prevenção & controle , Nanosferas , Trombocitopenia/complicações , Animais , Endotélio/metabolismo , Fibrinogênio/metabolismo , Hemorragia/metabolismo , Hemorragia/fisiopatologia , Camundongos , Agregação Plaquetária/efeitos dos fármacos , Análise de Sobrevida
17.
Blood ; 136(23): 2691-2702, 2020 12 03.
Artigo em Inglês | MEDLINE | ID: mdl-32659785

RESUMO

The mechanisms by which phlebotomy promotes the mobilization of hepatic iron stores are not well understood. NCOA4 (nuclear receptor coactivator 4) is a widely expressed intracellular protein previously shown to mediate the autophagic degradation of ferritin. Here, we investigate a local requirement for NCOA4 in the regulation of hepatic iron stores and examine mechanisms of NCOA4 regulation. Hepatocyte-targeted Ncoa4 knockdown in nonphlebotomized mice had only modest effects on hepatic ferritin subunit levels and nonheme iron concentration. After phlebotomy, mice with hepatocyte-targeted Ncoa4 knockdown exhibited anemia and hypoferremia similar to control mice with intact Ncoa4 regulation but showed a markedly impaired ability to lower hepatic ferritin subunit levels and hepatic nonheme iron concentration. This impaired hepatic response was observed even when dietary iron was limited. In both human and murine hepatoma cell lines, treatment with chemicals that stabilize hypoxia inducible factor (HIF), including desferrioxamine, cobalt chloride, and dimethyloxalylglycine, raised NCOA4 messenger RNA. This NCOA4 messenger RNA induction occurred within 3 hours, preceded a rise in NCOA4 protein, and was attenuated in the setting of dual HIF-1α and HIF-2α knockdown. In summary, we show for the first time that NCOA4 plays a local role in facilitating iron mobilization from the liver after blood loss and that HIF regulates NCOA4 expression in cells of hepatic origin. Because the prolyl hydroxylases that regulate HIF stability are oxygen- and iron-dependent enzymes, our findings suggest a novel mechanism by which hypoxia and iron deficiency may modulate NCOA4 expression to impact iron homeostasis.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Hemorragia/metabolismo , Hepatócitos/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Ferro/metabolismo , Fígado/metabolismo , Coativadores de Receptor Nuclear/biossíntese , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Linhagem Celular , Feminino , Regulação da Expressão Gênica , Técnicas de Silenciamento de Genes , Hemorragia/genética , Hemorragia/patologia , Hepatócitos/patologia , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Fígado/patologia , Camundongos , Coativadores de Receptor Nuclear/genética
18.
Eur J Pharmacol ; 883: 173310, 2020 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-32619674

RESUMO

Hemorrhagic cystitis is one of the most important complications of cyclophosphamide, a drug widely used in cancer chemotherapy and bone marrow transplantation. 5-HT3 antagonists are anti-emetic agents and have been shown to have notable anti-inflammatory and antioxidant properties. This study was designed to investigate the possible protective effects of tropisetron against cyclophosphamide-induced hemorrhagic cystitis in rats. Hemorrhagic cystitis was induced in female rats by cyclophosphamide (270 mg/kg). Tropisetron (2.5, 5 and 7.5 mg/kg), granisetron (2.5 and 5 mg/kg), and ondansetron (5 mg/kg) were injected 15 min before, 4 and 8 h after cyclophosphamide. To evaluate the role of alpha7 nicotinic acetylcholine receptor (α7nAChR), its antagonist, methyllycaconitine (5 mg/kg) was administered 30 min before tropisetron. After 24 h, animals were killed under anesthesia. Macroscopic and histological changes were evaluated. Malondialdehyde (MDA), glutathione (GSH) and Evans blue were measured spectrophotometrically. Furthermore, the protein levels of p38 mitogen-activated protein kinases (P38 MAPK), p-P38, signal transducer and activator of transcription 3 (STAT3), p-STAT3 and Poly (ADP-ribose) polymerase (PARP) were determined using Western blot. Cyclophosphamide administration significantly induced histopathological damages and increased MDA, p-p38/p38, p-STAT3/STAT3, and PARP levels compared with the saline group. Tropisetron treatment diminished histopathological injuries as well as MDA level, and STAT3 activity compared to cyclophosphamide treated rats. Co-administration of methyllycaconitine with tropisetron, partially or completely reversed the protective effects of tropisetron. Our results showed that prophylactic administration of tropisetron markedly ameliorated the cyclophosphamide-induced bladder hemorrhage and inflammation in rats. These effects of tropisetron were α7nAChR dependent.


Assuntos
Cistite/prevenção & controle , Hemorragia/prevenção & controle , Agonistas Nicotínicos/farmacologia , Tropizetrona/farmacologia , Bexiga Urinária/efeitos dos fármacos , Receptor Nicotínico de Acetilcolina alfa7/agonistas , Animais , Anti-Inflamatórios/farmacologia , Ciclofosfamida , Cistite/induzido quimicamente , Cistite/metabolismo , Cistite/patologia , Modelos Animais de Doenças , Feminino , Granisetron/farmacologia , Hemorragia/induzido quimicamente , Hemorragia/metabolismo , Hemorragia/patologia , Mediadores da Inflamação/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Ondansetron/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Fosforilação , Poli(ADP-Ribose) Polimerases/metabolismo , Ratos Wistar , Fator de Transcrição STAT3/metabolismo , Antagonistas do Receptor 5-HT3 de Serotonina/farmacologia , Transdução de Sinais , Bexiga Urinária/metabolismo , Bexiga Urinária/patologia , Receptor Nicotínico de Acetilcolina alfa7/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
19.
J Cell Mol Med ; 24(16): 9097-9100, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32588533

RESUMO

The BK polyomavirus (BKPyV) has pathogenic relevance especially in immunocompromised patients. No causal therapy has been established yet. Therefore, new therapeutic targets need to be identified in experimental studies. A 3D organotypic cell culture model with primary urothelial cells and fibroblasts was used as infection model. The detection of virus replication was performed with quantitative polymerase chain reaction (qPCR), and immunohistochemistry (IHC) was also used for analysis. Interleukin levels were measured by enzyme-linked immunosorbent assay (ELISA). Interestingly, the signal transducer and activator of transcription 3 (STAT3) pathway seems to be activated during infection with BKPyV, for example phosphorylated STAT3 is significantly (P < 0.0001) elevated on day 6 following infection. Therefore, we performed ELISAs for involved interleukins in STAT3 pathway. Interleukin 11 (IL-11) was significantly (P = 0.026) elevated at day 9. Subsequently, 3D cultures were treated with IL-11 neutralizing antibody. At day 9 following infection, the median virus replication rate is 4.4 × 106 copies/ml. The difference to replication rate without treatment was significantly lower at day 6 (P < 0.0001) and at day 9 (P < 0.0001), respectively. STAT3 pathways seem to be involved during BKPyV infection and need further investigation in experimental studies. A very promising target for treatment might be IL-11.


Assuntos
Vírus BK/patogenicidade , Hemorragia/metabolismo , Interleucina-11/metabolismo , Infecções por Polyomavirus/metabolismo , Vírus BK/genética , Técnicas de Cultura de Células/métodos , Células Cultivadas , Cistite , Fibroblastos/metabolismo , Fibroblastos/virologia , Hemorragia/virologia , Humanos , Infecções por Polyomavirus/virologia , Fator de Transcrição STAT3/metabolismo , Urotélio/metabolismo , Urotélio/virologia , Replicação Viral/genética
20.
J Clin Pharm Ther ; 45(4): 602-608, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32449992

RESUMO

WHAT IS KNOWN AND OBJECTIVE: Low-molecular-weight heparin (LMWH) is widely used in the prevention and treatment of venous thromboembolism (VTE), and anti-Xa assay is the gold standard for monitoring LMWH. However, it is still controversial whether monitoring is necessary for patients receiving LMWH therapy. Therefore, we conducted a meta-analysis to explore the effect of anti-Xa monitoring on the safety and efficacy of LMWH anticoagulant therapy. METHODS: PubMed, EMBASE, Web of Science and The Cochrane Library were searched on 27 May 2019 for eligible studies published in English. Odds ratio (OR) and 95% confidence intervals (CI) were estimated (Mantel-Haenszel method) using Review Manager version 5.3 software. The systematic review and meta-analysis was performed according to the recommendations of the Preferred Reporting Items of Systematic Reviews and Meta-Analyses (PRISMA) statement. RESULTS AND DISCUSSION: Six studies involving 1617 patients were eligible for our meta-analysis, with 724 patients in the anti-Xa monitoring group and 893 patients in the control group. There was no significant difference in the incidence of bleeding events between the two groups, while the anti-Xa monitoring group had a lower incidence of venous thromboembolism events (OR 0.44, 95% CI 0.29-0.68, P = .0002). Subgroup analysis showed that the incidence of venous thromboembolism events in the anti-Xa monitoring group was lower than that in the control group when the trough level was monitored (OR 0.40, 95% CI 0.25-0.63, P < .0001), while there was no significant difference between the two groups when the peak level was monitored. WHAT IS NEW AND CONCLUSION: Patients receiving LMWH anticoagulant therapy to prevent VTE can benefit from anti-Xa monitoring, for which the trough level may be the more appropriate time status to monitor.


Assuntos
Anticoagulantes/efeitos adversos , Anticoagulantes/uso terapêutico , Coagulação Sanguínea/efeitos dos fármacos , Inibidores do Fator Xa/uso terapêutico , Fator Xa/metabolismo , Heparina de Baixo Peso Molecular/efeitos adversos , Heparina de Baixo Peso Molecular/uso terapêutico , Hemorragia/induzido quimicamente , Hemorragia/metabolismo , Humanos , Razão de Chances , Tromboembolia Venosa/tratamento farmacológico , Tromboembolia Venosa/metabolismo
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