RESUMO
BACKGROUND: Patients with acetaminophen (APAP)-induced acute liver failure (ALF) display both hyper- and hypocoagulable changes not necessarily recapitulated by standard hepatotoxic doses of APAP used in mice (eg, 300 mg/kg). OBJECTIVES: We sought to examine coagulation activation in vivo and plasma coagulation potential ex vivo in experimental settings of APAP-induced hepatotoxicity and repair (300-450 mg/kg) and APAP-induced ALF (600 mg/kg) in mice. RESULTS: APAP-induced ALF was associated with increased plasma thrombin-antithrombin complexes, decreased plasma prothrombin, and a dramatic reduction in plasma fibrinogen compared with lower APAP doses. Hepatic fibrin(ogen) deposits increased independent of APAP dose, whereas plasma fibrin(ogen) degradation products markedly increased in mice with experimental ALF. Early pharmacologic anticoagulation (+2 hours after 600 mg/kg APAP) limited coagulation activation and reduced hepatic necrosis. The marked coagulation activation evident in mice with APAP-induced ALF was associated with a coagulopathy detectable ex vivo in plasma. Specifically, prolongation of the prothrombin time and inhibition of tissue factor-initiated clot formation were evident even after restoration of physiological fibrinogen concentrations. Plasma endogenous thrombin potential was similarly reduced at all APAP doses. Interestingly, in the presence of ample fibrinogen, â¼10 times more thrombin was required to clot plasma from mice with APAP-induced ALF compared with plasma from mice with simple hepatotoxicity. CONCLUSION: The results indicate that robust pathologic coagulation cascade activation in vivo and suppressed coagulation ex vivo are evident in mice with APAP-induced ALF. This unique experimental setting may fill an unmet need as a model to uncover mechanistic aspects of the complex coagulopathy of ALF.
Assuntos
Transtornos da Coagulação Sanguínea , Doença Hepática Induzida por Substâncias e Drogas , Falência Hepática , Camundongos , Animais , Acetaminofen/metabolismo , Trombina/metabolismo , Falência Hepática/metabolismo , Falência Hepática/patologia , Fígado/metabolismo , Fibrina/metabolismo , Transtornos da Coagulação Sanguínea/induzido quimicamente , Transtornos da Coagulação Sanguínea/metabolismo , Fibrinogênio/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/patologia , Camundongos Endogâmicos C57BLRESUMO
BACKGROUND: Blood coagulation protease activity is proposed to drive hepatic fibrosis through activation of protease-activated receptors (PARs). Whole-body PAR-1 deficiency reduces experimental hepatic fibrosis, and in vitro studies suggest a potential contribution by PAR-1 expressed by hepatic stellate cells. However, owing to a lack of specific tools, the cell-specific role of PAR-1 in experimental hepatic fibrosis has never been formally investigated. Using a novel mouse expressing a conditional PAR-1 allele, we tested the hypothesis that PAR-1 expressed by hepatic stellate cells contributes to hepatic fibrosis. METHODS: PAR-1flox/flox mice were crossed with mice expressing Cre recombinase controlled by the lecithin retinol acyltransferase (LRAT) promoter, which induces recombination in hepatic stellate cells. Male PAR-1flox/flox/LRATCre and PAR-1flox/flox mice were challenged twice weekly with carbon tetrachloride (CCl4, 1 mL/kg i.p.) for 6 weeks to induce liver fibrosis. RESULTS: PAR-1 mRNA levels were reduced (>95%) in hepatic stellate cells isolated from PAR-1flox/flox/LRATCre mice. Hepatic stellate cell activation was evident in CCl4-challenged PAR-1flox/flox mice, indicated by increased α-smooth muscle actin labeling and induction of several profibrogenic genes. CCl4-challenged PAR-1flox/flox mice displayed robust hepatic collagen deposition, indicated by picrosirius red staining and type I collagen immunolabeling. Notably, stellate cell activation and collagen deposition were significantly reduced (>30%) in PAR-1flox/flox/LRATCre mice. Importantly, the reduction in liver fibrosis was not a consequence of reduced acute CCl4 hepatotoxicity in PAR-1flox/flox/LRATCre mice. CONCLUSIONS: The results constitute the first direct experimental evidence that PAR-1 expressed by stellate cells directly promotes their profibrogenic phenotype and hepatic fibrosis in vivo.
RESUMO
BACKGROUND & AIM: Acetaminophen (APAP)-induced acute liver failure is associated with substantial alterations in the hemostatic system. In mice, platelets accumulate in the liver after APAP overdose and appear to promote liver injury. Interestingly, patients with acute liver injury have highly elevated levels of the platelet-adhesive protein von Willebrand factor (VWF), but a mechanistic connection between VWF and progression of liver injury has not been established. We tested the hypothesis that VWF contributes directly to experimental APAP-induced acute liver injury. METHODS: Wild-type mice and VWF-deficient (Vwf-/-) mice were given a hepatotoxic dose of APAP (300â¯mg/kg, i.p.) or vehicle (saline). VWF plasma levels were measured by ELISA, and liver necrosis or hepatocyte proliferation was measured by immunohistochemistry. Platelet and VWF deposition were measured by immunofluorescence. RESULTS: In wild-type mice, VWF plasma levels, high molecular weight (HMW) VWF multimers, and VWF activity decreased 24â¯h after APAP challenge. These changes coupled to robust hepatic VWF and platelet deposition, although VWF deficiency had minimal effect on peak hepatic platelet accumulation or liver injury. VWF plasma levels were elevated 48â¯h after APAP challenge, but with relative reductions in HMW multimers and VWF activity. Whereas hepatic platelet aggregates persisted in livers of APAP-challenged wild-type mice, platelets were nearly absent in Vwf-/- mice 48â¯h after APAP challenge. The absence of platelet aggregates was linked to dramatically accelerated repair of the injured liver. Complementing observations in Vwf-/- mice, blocking VWF or the platelet integrin αIIbß3 during development of injury significantly reduced hepatic platelet aggregation and accelerated liver repair in APAP-challenged wild-type mice. CONCLUSION: These studies are the first to suggest a mechanistic link between VWF, hepatic platelet accumulation, and liver repair. Targeting VWF might provide a novel therapeutic approach to improve repair of the APAP-injured liver. LAY SUMMARY: Patients with acute liver injury due to acetaminophen overdose have highly elevated levels of the platelet-adhesive protein von Willebrand factor. It is not known whether von Willebrand factor plays a direct role in the progression of acute liver injury. We discovered that von Willebrand factor delays repair of the acetaminophen-injured liver in mice and that targeting von Willebrand factor, even in mice with established liver injury, accelerates liver repair.
Assuntos
Acetaminofen/efeitos adversos , Analgésicos não Narcóticos/efeitos adversos , Plaquetas/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/sangue , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Fígado/metabolismo , Fator de von Willebrand/metabolismo , Acetaminofen/administração & dosagem , Analgésicos não Narcóticos/administração & dosagem , Animais , Coagulação Sanguínea/efeitos dos fármacos , Humanos , Fígado/patologia , Masculino , Taxa de Depuração Metabólica , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Necrose , Agregação Plaquetária/efeitos dos fármacos , Fator de von Willebrand/administração & dosagem , Fator de von Willebrand/genética , Fator de von Willebrand/farmacocinéticaRESUMO
A 7-month-old Miniature Poodle acquired from a pet store developed cough and subsequently respiratory distress compatible with Bordetella bronchiseptica infection. Partial but incomplete resolution of clinical signs and thoracic radiographic/computed tomographic imaging lesions were noted with use of susceptibility-guided antimicrobials. Additionally, a concern for an infectious nidus led to left cranial lung lobectomy at 9 months of age. Histopathology predominantly revealed polypoid and constrictive bronchiolitis obliterans (i.e., small airway disease). Intermittent antimicrobial administration over the next 5 years failed to blunt progressive clinical signs. At 8 years, necropsy confirmed severe airway-centered interstitial fibrosis. This pattern of fibrosis was strongly suggestive of underlying small airway disease as the trigger. In retrospect, post-infectious bronchiolitis obliterans (PIBO), a syndrome in young children caused by pulmonary infections but not yet recognized in pet dogs, likely initiated a pathway of fibrosis in this dog. In dogs with risk factors for community-acquired pathogens such as Bordetella bronchiseptica, PIBO is a differential diagnosis with development of severe, persistent respiratory signs incompletely responsive to appropriate antimicrobials. Untreated PIBO may lead to airway-centered interstitial fibrosis. Future study is required to determine if targeted therapy of PIBO could alter the course of end-stage pulmonary fibrosis.
RESUMO
A 2 yr old, neutered male rottweiler was evaluated for a chronic cough that had acutely worsened. Computed tomographic examination revealed a diffuse alveolar pattern in the right, middle, and left cranial lung lobes. Aerated parenchymal tissue was not observed in the left cranial lung lobe, and both lobes were markedly decreased in volume. Lobectomy of the right middle and left cranial lung lobes was performed. Histopathologic examination of both lungs identified alveolar collapse associated with marked chronic bronchial and bronchiolar luminal concentric fibrosis leading to reduced airway lumen diameter and bronchiolar destruction. The clinical signs and airway pathology were consistent with constrictive bronchiolitis obliterans. The dog remained stable for over 2 yr with glucocorticoid therapy and intermittent antimicrobics. Although the polypoid form of bronchiolitis obliterans has been described in cattle and occasionally in dogs, constrictive bronchiolitis obliterans has not been reported previously in veterinary species.
Assuntos
Bronquiolite Obliterante/veterinária , Doenças do Cão/diagnóstico , Animais , Antibacterianos/uso terapêutico , Bronquiolite Obliterante/diagnóstico , Bronquiolite Obliterante/tratamento farmacológico , Constrição Patológica , Doenças do Cão/tratamento farmacológico , Cães , Masculino , Prednisona/uso terapêuticoAssuntos
Infecções por Pasteurellaceae/veterinária , Pasteurellaceae/isolamento & purificação , Doenças dos Ovinos/patologia , Animais , Evolução Fatal , Feminino , Infecções por Pasteurellaceae/microbiologia , Infecções por Pasteurellaceae/patologia , Ovinos , Doenças dos Ovinos/diagnóstico , Doenças dos Ovinos/microbiologiaRESUMO
Equine herpesvirus 5 (EHV-5) infection is associated with pulmonary fibrosis in horses, but further studies on EHV-5 persistence in equine cells are needed to fully understand viral and host contributions to disease pathogenesis. Our aim was to develop a quantitative PCR (qPCR) assay to measure EHV-5 viral copy number in equine cell cultures, blood lymphocytes, and nasal swabs of horses. Furthermore, we used a recently developed equine primary respiratory cell culture system to study EHV-5 pathogenesis at the respiratory tract. PCR primers and a probe were designed to target gene E11 of the EHV-5 genome. Sensitivity and repeatability were established, and specificity was verified by testing multiple isolates of EHV-5, as well as DNA from other equine herpesviruses. Four-week old fully differentiated (mature), newly seeded (immature) primary equine respiratory epithelial cell (ERECs), and equine dermal cell cultures were inoculated with EHV-5 and the cells and supernatants collected daily for 14days. Blood lymphocytes and nasal swabs were collected from horses experimentally infected with equine herpesvirus 1 (EHV-1). The qPCR assay detected EHV-5 at stable concentrations throughout 14days in inoculated mature EREC and equine dermal cell cultures (peaking at 202 and 5861 viral genomes per 106 cellular ß actin, respectively). EHV-5 copies detected in the immature EREC cultures increased over 14days and reached levels greater than 10,000 viral genomes per 106 cellular ß actin. Moreover, EHV-5 was detected in the lymphocytes of 76% of horses and in the nasal swabs of 84% of horses experimentally infected with EHV-1 pre-inoculation with EHV-1. Post-inoculation with EHV-1, EHV-5 was detected in lymphocytes of 52% of horses while EHV-5 levels in nasal swabs were not significantly different from pre-inoculation levels. In conclusion, qPCR was a reliable technique to investigate viral load in in vivo and in vitro samples, and EHV-5 replication in equine epithelial cells may be influenced by cellular stages of differentiation.
Assuntos
Gammaherpesvirinae/isolamento & purificação , Gammaherpesvirinae/fisiologia , Infecções por Herpesviridae/veterinária , Doenças dos Cavalos/virologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Replicação Viral , Animais , Replicação do DNA , DNA Viral/genética , Células Epiteliais/virologia , Gammaherpesvirinae/genética , Infecções por Herpesviridae/virologia , Herpesvirus Equídeo 1/isolamento & purificação , Cavalos , Linfócitos/virologia , Nariz/virologia , Sistema Respiratório/virologia , Carga ViralRESUMO
Trichloroethylene (TCE) is a persistent environmental contaminant proposed to contribute to autoimmune disease. Experimental studies in lupus-prone MRL+/+ mice have suggested that TCE exposure can trigger autoimmune hepatitis. The vast majority of studies examining the connection between TCE and autoimmunity utilize this model, and the impact of TCE exposure in other established models of autoimmune liver disease is not known. We tested the hypothesis that TCE exposure exacerbates experimental hepatic autoimmunity in dominant negative transforming growth factor beta receptor type II (dnTGFBRII) mice, which develop serological and histological features resembling human primary biliary cholangitis. Female 8-week-old wild-type and dnTGFBRII mice were exposed to TCE (0.5 mg/ml) or vehicle (1% ethoxylated castor oil) in the drinking water for 12 or 22 weeks. Liver histopathology in 20- and 30-week-old wild-type mice was unremarkable irrespective of treatment. Mild portal inflammation was observed in vehicle-exposed 20-week-old dnTGFBRII mice and was not exacerbated by TCE exposure. Vehicle-exposed 30-week-old dnTGFBRII mice developed anti-mitochondrial antibodies, marked hepatic inflammation with necrosis, and hepatic accumulation of both B and T lymphocytes. To our surprise, TCE exposure dramatically reduced hepatic parenchymal inflammation and injury in 30-week-old dnTGFBRII mice, reflected by changes in hepatic proinflammatory gene expression, serum chemistry, and histopathology. Interestingly, TCE did not affect hepatic B cell accumulation or induction of the anti-inflammatory cytokine IL10. These data indicate that TCE exposure reduces autoimmune liver injury in female dnTGFBRII mice and suggests that the precise effect of environmental chemicals in autoimmunity depends on the experimental model.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas/imunologia , Colangite/imunologia , Modelos Animais de Doenças , Hepatite Autoimune/imunologia , Tricloroetileno/toxicidade , Animais , Autoanticorpos/sangue , Doença Hepática Induzida por Substâncias e Drogas/genética , Doença Hepática Induzida por Substâncias e Drogas/patologia , Colangite/genética , Colangite/patologia , Feminino , Interação Gene-Ambiente , Hepatite Autoimune/genética , Hepatite Autoimune/patologia , Masculino , Camundongos Transgênicos , Proteínas Serina-Treonina Quinases/genética , Receptor do Fator de Crescimento Transformador beta Tipo II , Receptores de Fatores de Crescimento Transformadores beta/genéticaRESUMO
Regional differences in large equine pulmonary artery reactivity exist. It is not known if this heterogeneity extends into small vessels. The hypothesis that there is regional heterogeneity in small pulmonary artery and vein reactivity to sympathomimetics (phenylephrine and isoproterenol) and a parasympathomimetic (methacholine) was tested using wire myography on small vessels from caudodorsal (CD) and cranioventral (CV) lung of 12 horses [9 mares, 3 geldings, 8.67 ± 0.81 (age ± SE) yr, of various breeds that had never raced]. To study relaxation, vessels were precontracted with U46619 (10(-6) M). Methacholine mechanism of action was investigated using L-nitroarginine methylester (L-NAME, 100 µM) and indomethacin (10 µM). Phenylephrine did not contract any vessels. Isoproterenol relaxed CD arteries more than CV arteries (maximum relaxation 28.18% and 48.67%; Log IC50 ± SE -7.975 ± 0.1327 and -8.033 ± 0.1635 for CD and CV, respectively, P < 0.0001), but not veins. Methacholine caused contraction of CD arteries (maximum contraction 245.4%, Log EC50 ± SE -6.475 ± 0.3341), and relaxation of CV arteries (maximum relaxation 40.14%, Log IC50 ± SE -6.791 ± 0.1954) and all veins (maximum relaxation 50.62%, Log IC50 ± SE -6.932 ± 0.1986) in a nonregion-dependent manner. L-NAME (n = 8, P < 0.0001) and indomethacin (n = 7, P < 0.0001) inhibited methacholine-induced relaxation of CV arteries, whereas indomethacin augmented CD artery contraction (n = 8, P < 0.0001). Our data demonstrate significant regional heterogeneity in small blood vessel reactivity when comparing the CD to the CV region of the equine lung.
Assuntos
Pulmão/irrigação sanguínea , Artéria Pulmonar/fisiologia , Veias/fisiologia , Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico/farmacologia , Animais , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/fisiologia , Feminino , Cavalos , Indometacina/farmacologia , Isoproterenol/farmacologia , Masculino , Cloreto de Metacolina/farmacologia , Miografia/métodos , NG-Nitroarginina Metil Éster/farmacologia , Fenilefrina/farmacologia , Artéria Pulmonar/efeitos dos fármacos , Vasoconstrição/efeitos dos fármacos , Vasoconstritores/farmacologia , Vasodilatação/efeitos dos fármacos , Vasodilatação/fisiologia , Veias/efeitos dos fármacosRESUMO
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a potent aryl hydrocarbon receptor agonist that elicits dose-dependent hepatic fat accumulation and inflammation that can progress to steatohepatitis. To investigate intestine-liver interactions that contribute to TCDD-elicited steatohepatitis, we examined the dose-dependent effects of TCDD (0.01, 0.03, 0.1, 0.3, 1, 3, 10, or 30 µg/kg) on jejunal epithelial gene expression in C57BL/6 mice orally gavaged every 4 days for 28 days. Agilent 4x44K whole-genome microarray analysis of the jejunal epithelium identified 439 differentially expressed genes (|fold change| ≥ 1.5, P1(t) ≥ 0.999) across 1 or more doses, many related to lipid metabolism and immune system processes. TCDD-elicited differentially expressed genes were associated with lipolysis, fatty acid/cholesterol absorption and transport, the Kennedy pathway, and retinol metabolism, consistent with increased hepatic fat accumulation. Moreover, several major histocompatibility complex (MHC) class II genes (H2-Aa, H2-Ab1, H2-DMb1, Cd74) were repressed, coincident with decreased macrophage and dendritic cell levels in the lamina propria, suggesting migration of antigen-presenting cells out of the intestine. In contrast, hepatic RNA-Seq analysis identified increased expression of MHC class II genes, as well as chemokines and chemokine receptors involved in macrophage recruitment (Ccr1, Ccr5, Ccl5, Cx3cr1), consistent with hepatic F4/80 labeling and macrophage infiltration into the liver. Collectively, these results suggest TCDD elicits changes that support hepatic lipid accumulation, macrophage migration, and the progression of hepatic steatosis to steatohepatitis.
Assuntos
Poluentes Ambientais/toxicidade , Imunidade nas Mucosas/efeitos dos fármacos , Mucosa Intestinal/efeitos dos fármacos , Jejuno/efeitos dos fármacos , Metabolismo dos Lipídeos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Hepatopatia Gordurosa não Alcoólica/induzido quimicamente , Dibenzodioxinas Policloradas/toxicidade , Animais , Relação Dose-Resposta a Droga , Feminino , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica , Genes MHC da Classe II , Imunidade nas Mucosas/genética , Mucosa Intestinal/imunologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Jejuno/imunologia , Jejuno/metabolismo , Jejuno/patologia , Metabolismo dos Lipídeos/genética , Fígado/metabolismo , Fígado/patologia , Camundongos Endogâmicos C57BL , Hepatopatia Gordurosa não Alcoólica/genética , Hepatopatia Gordurosa não Alcoólica/imunologia , Hepatopatia Gordurosa não Alcoólica/metabolismo , Hepatopatia Gordurosa não Alcoólica/patologia , Medição de Risco , Fatores de TempoRESUMO
Exercise-induced pulmonary hemorrhage is a performance-limiting condition of racehorses associated with severe pathology, including small pulmonary vein remodeling. Pathology is limited to caudodorsal (CD) lung. Mechanical properties of equine pulmonary microvasculature have not been studied. We hypothesized that regional differences in pulmonary artery and vein mechanical characteristics do not exist in control animals, and that racing and venous remodeling impact pulmonary vein mechanical properties in CD lung. Pulmonary arteries and veins [range of internal diameters 207-386 ± 67 µm (mean ± SD)] were harvested from eight control and seven raced horses. With the use of wire myography, CD and cranioventral (CV) vessels were stretched in 10-µm increments. Peak wall tension was plotted against changes in diameter (length). Length-tension data were compared between vessel type, lung region, and horse status (control and raced). Pulmonary veins are stiffer walled than arteries. CD pulmonary arteries are stiffer than CV arteries, whereas CV veins are stiffer than CD veins. Racing is associated with increased stiffness of CD pulmonary veins and, to a lesser extent, CV arteries. For example, at 305 µm, tension in raced and control CD veins is 27.74 ± 2.91 and 19.67 ± 2.63 mN/mm (means ± SE; P < 0.05, Bonferroni's multiple-comparisons test after two-way ANOVA), and 16.12 ± 2.04 and 15.07 ± 2.47 mN/mm in raced and control CV arteries, respectively. This is the first report of an effect of region and/or exercise on mechanical characteristics of small pulmonary vessels. These findings may implicate pulmonary vein remodeling in exercise-induced pulmonary hemorrhage pathogenesis.
Assuntos
Traumatismos em Atletas/patologia , Traumatismos em Atletas/veterinária , Doenças dos Cavalos/patologia , Cavalos , Pulmão/patologia , Microcirculação , Circulação Pulmonar , Animais , Feminino , Pulmão/fisiopatologia , Masculino , Miografia , Condicionamento Físico Animal , Artéria Pulmonar/patologia , Veias Pulmonares/patologia , Rigidez VascularRESUMO
We examined the effects of two over-the-counter H1-antihistamines on the progression of fatty liver disease in male C57Bl/6 wild-type and apolipoprotein E (ApoE)-/- mice. Mice were fed a high-fat diet (HFD) for 3 mo, together with administration of either cetirizine (4 mg/kg body wt) or fexofenadine (40 mg/kg body wt) in drinking water. Antihistamine treatments increased body weight gain, gonadal fat deposition, liver weight, and hepatic steatosis in wild-type mice but not in ApoE-/- mice. Lobular inflammation, acute inflammation, and necrosis were not affected by H1-antihistamines in either genotype. Serum biomarkers of liver injury tended to increase in antihistamine-treated wild-type mice. Serum level of glucose was increased by fexofenadine, whereas lipase was increased by cetirizine. H1-antihistamines reduced the mRNA expression of ApoE and carbohydrate response element-binding protein in wild-type mice, without altering the mRNA expression of sterol regulatory element-binding protein 1c, fatty acid synthase, or ApoB100, in either genotype. Fexofenadine increased both triglycerides and cholesterol ester, whereas cetirizine increased only cholesterol ester in liver, with a concomitant decrease in serum triglycerides by both antihistamines in wild-type mice. Antihistamines increased hepatic levels of conjugated bile acids in wild-type mice, with the effect being significant in fexofenadine-treated animals. The increase was associated with changes in the expression of organic anion transport polypeptide 1b2 and bile salt export pump. These results suggest that H1-antihistamines increase the progression of fatty liver disease in wild-type mice, and there seems to be an association between the severity of disease, presence of ApoE, and increase in hepatic bile acid levels.
Assuntos
Apolipoproteínas E/deficiência , Cetirizina/toxicidade , Dieta Hiperlipídica , Fígado Gorduroso/induzido quimicamente , Antagonistas dos Receptores Histamínicos H1/toxicidade , Fígado/efeitos dos fármacos , Terfenadina/análogos & derivados , Membro 11 da Subfamília B de Transportadores de Cassetes de Ligação de ATP , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Animais , Apolipoproteínas E/genética , Ácidos e Sais Biliares/metabolismo , Biomarcadores/sangue , Ésteres do Colesterol/metabolismo , Modelos Animais de Doenças , Fígado Gorduroso/sangue , Fígado Gorduroso/genética , Fígado Gorduroso/patologia , Regulação da Expressão Gênica , Lipogênese/efeitos dos fármacos , Lipogênese/genética , Fígado/metabolismo , Fígado/patologia , Transportador 1 de Ânion Orgânico Específico do Fígado , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Transportadores de Ânions Orgânicos Sódio-Independentes/genética , Transportadores de Ânions Orgânicos Sódio-Independentes/metabolismo , Índice de Gravidade de Doença , Terfenadina/toxicidade , Triglicerídeos/metabolismoRESUMO
Hepatic fibrin deposition has been shown to inhibit hepatocellular injury in mice exposed to the bile duct toxicant α-naphthylisothiocyanate (ANIT). Degradation of fibrin clots by fibrinolysis controls the duration and extent of tissue fibrin deposition. Thus, we sought to determine the effect of treatment with the antifibrinolytic drug tranexamic acid (TA) and plasminogen activator inhibitor-1 (PAI-1) deficiency on ANIT-induced liver injury and fibrosis in mice. Plasmin-dependent lysis of fibrin clots was impaired in plasma from mice treated with TA (1200 mg/kg i.p., administered twice daily). Prophylactic TA administration reduced hepatic inflammation and hepatocellular necrosis in mice fed a diet containing 0.025% ANIT for 2 weeks. Hepatic type 1 collagen mRNA expression and deposition increased markedly in livers of mice fed ANIT diet for 4 weeks. To determine whether TA treatment could inhibit this progression of liver fibrosis, mice were fed ANIT diet for 4 weeks and treated with TA for the last 2 weeks. Interestingly, TA treatment largely prevented increased deposition of type 1 collagen in livers of mice fed ANIT diet for 4 weeks. In contrast, biliary hyperplasia/inflammation and liver fibrosis were significantly increased in PAI-1(-/-) mice fed ANIT diet for 4 weeks. Overall, the results indicate that fibrinolytic activity contributes to ANIT diet-induced liver injury and fibrosis in mice. In addition, these proof-of-principle studies suggest the possibility that therapeutic intervention with an antifibrinolytic drug could form a novel strategy to prevent or reduce liver injury and fibrosis in patients with liver disease.
Assuntos
Antifibrinolíticos/uso terapêutico , Doenças dos Ductos Biliares/tratamento farmacológico , Cirrose Hepática/prevenção & controle , Fígado/efeitos dos fármacos , Ácido Tranexâmico/uso terapêutico , 1-Naftilisotiocianato/farmacologia , Animais , Antifibrinolíticos/administração & dosagem , Doenças dos Ductos Biliares/induzido quimicamente , Doenças dos Ductos Biliares/metabolismo , Doenças dos Ductos Biliares/patologia , Colágeno Tipo I/biossíntese , Modelos Animais de Doenças , Fibrina/metabolismo , Fibrinogênio/genética , Fígado/metabolismo , Fígado/patologia , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/metabolismo , Cirrose Hepática/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Inibidor 1 de Ativador de Plasminogênio/deficiência , Inibidor 1 de Ativador de Plasminogênio/genética , Ácido Tranexâmico/administração & dosagemRESUMO
BACKGROUND: The clinical outcome of idiopathic pulmonary fibrosis (IPF) is poor, with a 50% survival rate at 3 years. Furthermore, current treatments provide little amelioration of symptoms. Despite significant advances in understanding the clinical features and pathobiology of IPF, further advances have been hampered by a lack of suitable animal models of the disease. Interestingly, spontaneously occurring disorders with a similarity to IPF have been recognized in the dog, cat, horse, and donkey. These disorders share clinical and pathologic features with human IPF and are emerging diseases of veterinary importance. PURPOSE: To improve awareness about these disorders in domestic animals and stimulate interactions between disciplines, and to facilitate the elucidation of mechanisms of fibrosing lung disorders using a comparative natural-occurrence disease model approach. METHODS: A 1-day meeting joined physicians, veterinarians, pathologists, researchers, and advocacy experts to discuss information available in this area. A review of the literature was conducted, and an executive committee discussed the findings and prepared a summary statement during subsequent meetings. RESULTS: Clinical, diagnostic, and treatment opportunities were identified, and common areas of interest where collaborative efforts could accelerate discovery regarding etiological factors, methods for early detection, determinants of disease progression, and novel therapies were defined. CONCLUSIONS: Comparing fibrosing lung disorders in humans and domestic animals will allow for a better understanding of the similarities and differences among species and may offer novel insights into the underlying mechanisms of spontaneously occurring fibrotic lung diseases.
Assuntos
Pulmão/patologia , Fibrose Pulmonar/patologia , Pneumologia , Sociedades Médicas , Animais , Animais Domésticos , Gatos , Cães , Histologia Comparada , Humanos , Fibrose Pulmonar Idiopática/diagnóstico por imagem , Fibrose Pulmonar Idiopática/patologia , Fibrose Pulmonar Idiopática/veterinária , Pulmão/diagnóstico por imagem , Fibrose Pulmonar/diagnóstico por imagem , Fibrose Pulmonar/veterinária , Tomografia Computadorizada por Raios X , Medicina VeterináriaRESUMO
Gammaherpesviruses (γHV) are implicated in the pathogenesis of pulmonary fibrosis in humans and murine models of lung fibrosis, however there is little direct experimental evidence that such viruses induce lung fibrosis in the natural host. The equine γHV EHV 5 is associated with equine multinodular pulmonary fibrosis (EMPF), a progressive fibrosing lung disease in its natural host, the horse. Experimental reproduction of EMPF has not been attempted to date. We hypothesized that inoculation of EHV 5 isolated from cases of EMPF into the lungs of clinically normal horses would induce lung fibrosis similar to EMPF. Neutralizing antibody titers were measured in the horses before and after inoculation with EHV 5. PCR and virus isolation was used to detect EHV 5 in antemortem blood and BAL samples, and in tissues collected postmortem. Nodular pulmonary fibrosis and induction of myofibroblasts occurred in EHV 5 inoculated horses. Mean lung collagen in EHV 5 inoculated horses (80 µg/mg) was significantly increased compared to control horses (26 µg/mg) (p < 0.5), as was interstitial collagen (32.6% ± 1.2% vs 23% ± 1.4%) (mean ± SEM; p < 0.001). Virus was difficult to detect in infected horses throughout the experiment, although EHV 5 antigen was detected in the lung by immunohistochemistry. We conclude that the γHV EHV 5 can induce lung fibrosis in the horse, and hypothesize that induction of fibrosis occurs while the virus is latent within the lung. This is the first example of a γHV inducing lung fibrosis in the natural host.
Assuntos
Gammaherpesvirinae/patogenicidade , Doenças dos Cavalos/virologia , Fibrose Pulmonar/virologia , Animais , Anticorpos Antivirais/imunologia , Gammaherpesvirinae/imunologia , Doenças dos Cavalos/imunologia , Doenças dos Cavalos/metabolismo , Cavalos , Fibrose Pulmonar/imunologia , Fibrose Pulmonar/metabolismoRESUMO
1,1,1-Trichloro-2,2-bis(2-chlorophenyl-4-chlorophenyl)ethane (o,p'-DDT) is an organochlorine pesticide and endocrine disruptor known to activate the estrogen receptor. Comprehensive ligand- and species-comparative dose- and time-dependent studies were conducted to systematically assess the uterine physiological, morphological and gene expression responses elicited by o,p'-DDT and ethynyl estradiol (EE) in immature ovariectomized C57BL/6 mice and Sprague-Dawley rats. Custom cDNA microarrays were used to identify conserved and divergent differential gene expression responses. A total of 1256 genes were differentially expressed by both ligands in both species, 559 of which exhibited similar temporal expression profiles suggesting that o,p'-DDT elicits estrogenic effects at high doses when compared to EE. However, 51 genes exhibited species-specific uterine expression elicited by o,p'-DDT. For example, carbonic anhydrase 2 exhibited species- and ligand-divergent expression as confirmed by quantitative real-time PCR. The identification of comparable temporal phenotypic responses linked to gene expression demonstrates that systematic comparative gene expression assessments are valuable for elucidating conserved and divergent estrogen signaling mechanisms in rodent uterotrophy.
Assuntos
DDT/toxicidade , Expressão Gênica , Praguicidas/toxicidade , Útero/efeitos dos fármacos , Animais , Disruptores Endócrinos/toxicidade , Estrogênios/toxicidade , Feminino , Perfilação da Expressão Gênica , Camundongos , Camundongos Endogâmicos C57BL , Análise de Sequência com Séries de Oligonucleotídeos , RNA/genética , RNA/isolamento & purificação , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Estrogênio/genética , Receptores de Estrogênio/metabolismo , Especificidade da Espécie , Útero/metabolismo , Útero/patologiaRESUMO
OBJECTIVE: To determine the effects of 2 weeks of intense exercise on expression of markers of pulmonary venous remodeling in the caudodorsal and cranioventral regions of the lungs of horses. ANIMALS: 6 horses. PROCEDURES: Tissue samples of the caudodorsal and cranioventral regions of lungs were obtained before and after conditioning and 2 weeks of intense exercise. Pulmonary veins were isolated, and a quantitative real-time PCR assay was used to determine mRNA expression of matrix metalloproteinase-2 and -9, tissue inhibitor of metalloproteinase-1 and -2, collagen type I, tenascin-C, endothelin-1, platelet-derived growth factor, transforming growth factor (TGF)-ß, and vascular endothelial growth factor (VEGF). Protein expression of collagen (via morphometric analysis) and tenascin-C, TGF-ß, and VEGF (via immunohistochemistry) was determined. RESULTS: Exercise-induced pulmonary hemorrhage was detected in 2 horses after exercise. The mRNA expression of matrix metalloproteinase-2 and -9, tissue inhibitor of metalloproteinase-2, TGF-ß, and VEGF was significantly lower in pulmonary veins obtained after exercise versus those obtained before exercise for both the caudodorsal and cranioventral regions of the lungs. Collagen content was significantly higher in tissue samples obtained from the caudodorsal regions of the lungs versus content in samples obtained from the cranioventral regions of the lungs both before and after exercise. Exercise did not alter protein expression of tenascin-C, TGF-ß, or VEGF. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this study indicated 2 weeks of intense exercise did not alter expression of marker genes in a manner expected to favor venous remodeling. Pulmonary venous remodeling is complex, and > 2 weeks of intense exercise may be required to induce such remodeling.
Assuntos
Cavalos/fisiologia , Pulmão/fisiologia , Condicionamento Físico Animal , Veias Pulmonares/fisiologia , Animais , Colágeno Tipo I/genética , Colágeno Tipo I/fisiologia , Feminino , Imuno-Histoquímica/veterinária , Masculino , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/fisiologia , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/fisiologia , RNA Mensageiro/química , RNA Mensageiro/genética , Distribuição Aleatória , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/fisiologia , Inibidor Tecidual de Metaloproteinase-2/genética , Inibidor Tecidual de Metaloproteinase-2/fisiologia , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/fisiologia , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/fisiologiaRESUMO
CASE DESCRIPTION: An 8-year-old 38-kg (84-lb) castrated male German Shepherd Dog cross was evaluated because of respiratory distress secondary to pneumothorax (detected radio-graphically prior to referral). CLINICAL FINDINGS: CT of the thorax confirmed the presence of pneumothorax and revealed pulmonary blebs without evidence of infiltrative pulmonary changes. A tentative diagnosis of primary spontaneous pneumothorax was made. TREATMENT AND OUTCOME: Exploratory median sternotomy revealed emphysematous changes along the margins of all lung lobes, with the ventral margins of the left cranial, right cranial, and right middle lung lobes most affected. Partial lobectomies of the ventral aspects of these lobes were performed. Histologic examination of tissue samples from the lung lobes revealed diffuse smooth muscle hypertrophy of the terminal and respiratory bronchioles with moderate numbers of peribronchiolar eosinophils. Mucus plugs and mucous cell metaplasia within the airway epithelium were also evident. After surgery, clinical signs resolved and the dog was discharged from the hospital 2 days later. Eight months after surgery, the dog developed a mild cough, and treatment with prednisolone (tapering dosage starting at 0.5 mg/kg [0.023 mg/lb], PO, q 12 h) was initiated. Dosage reduction resulted in recurrence of coughing; however, with continued prednisolone treatment at a dosage of 0.5 mg/kg, PO, once daily, the dog was not coughing at 10 months after surgery. CLINICAL RELEVANCE: Reactive bronchopneumopathy should be included as a differential diagnosis for spontaneous pneumothorax in dogs.