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BACKGROUND: The effectiveness of negative-pressure wound therapy (NPWT) in skin graft fixation has been demonstrated in several clinical studies. However, in vitro and in vivo studies on skin graft fixation with NPWT have been scarce. In this in vivo study, we aimed to determine whether NPWT fixation enhances skin graft survival and how it contributes to improving skin graft survival biologically. MATERIALS AND METHODS: We harvested skin from the bilateral abdominal wall of 88 mice after anesthetizing them. Full-thickness skin grafts (FTSGs) were performed on contralateral harvest sites, and grafts were fixed using NPWT (continuous and intermittent modes), conventional compression methods, and wrapping with polyurethane foam as a control group. On days 5 and 10 of grafting, the survival rates of the FTSGs were evaluated. Immunohistopathological analysis and measurement of the expression levels of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (FGF-2), and epidermal growth factor (EGF) were performed. RESULTS: The survival rates of FTSG in the continuous NPWT group were significantly higher than those in the other groups. The number of capillaries in the dermis was significantly higher in the continuous NPWT group than in the other groups. In the wound bed, VEGF levels were significantly higher in both NPWT groups than in the other groups. CONCLUSION: Continuous NPWT increases the survival rate of FTSGs and shortens the duration of skin graft survival.
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Supervivencia de Injerto , Terapia de Presión Negativa para Heridas , Trasplante de Piel , Terapia de Presión Negativa para Heridas/métodos , Trasplante de Piel/métodos , Animales , Supervivencia de Injerto/fisiología , Ratones , Masculino , Cicatrización de Heridas/fisiología , Factor A de Crecimiento Endotelial Vascular/metabolismo , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Factor de Crecimiento Epidérmico/metabolismo , Factores de Tiempo , Piel/patologíaRESUMEN
Lymphatic ascites following pelvic and para-aortic lymphadenectomy is a well-known complication. Surgical treatment and interventional radiology are required in a few cases. To determine the appropriate treatment strategy, it is important to preoperatively detect the presence and location of lymphatic leakage. However, the methods have yet to be established. We report a case in which lymphoscintigraphy with single-photon emission computerized tomography/computed tomography (SPECT/CT) was performed to evaluate pelvic lymphorrhea that occurred following total hysterectomy with pelvic and para-aortic lymphadenectomy for stage IIIA uterine sarcoma. Lymphoscintigraphy with SPECT/CT showed leakage of radioisotopes into the pelvic space, and intranodal lymphangiography was performed based on these findings. Following the procedure, the pelvic lymphorrhea improved, and no radioisotope leakage was confirmed by re-evaluation with lymphoscintigraphy with SPECT/CT. Our case indicates that lymphoscintigraphy with SPECT/CT may be useful for detecting the precise site of lymphatic leakage before interventional radiology or surgery.
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Negative-pressure wound therapy (NPWT) promotes wound healing by applying negative pressure to the wound surface. A quarter of a century after its introduction, NPWT has been used in various clinical conditions, although molecular biological evidence is insufficient due to delay in basic research. Here, we have summarized the history of NPWT, its mechanism of action, what is currently known about it, and what is expected to be known in the future. Particularly, attention has shifted from the four main mechanisms of NPWT to the accompanying secondary effects, such as effects on various cells, bacteria, and surgical wounds. This chapter will help the reader to understand the current status and shortcomings of NPWT-related research, which could aid in the development of basic research and, eventually, clinical use with stronger scientific evidence.
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Terapia de Presión Negativa para Heridas , Humanos , Vendajes , Cicatrización de Heridas , Infección de la Herida QuirúrgicaRESUMEN
BACKGROUND: Although the clinical efficacy of negative-pressure wound therapy (NPWT) is well known, many of its molecular biological mechanisms remain unresolved, mainly due to the difficulty and paucity of relevant in vitro studies. We attempted to develop an in vitro cell culture system capable of real-time monitoring of cells during NPWT treatment. MATERIALS AND METHODS: A novel negative-pressure cell culture system was developed by combining an inverted microscope, a stage-top incubator, a sealed metal chamber for cell culture, and an NPWT treatment device. Human keratinocytes, PSVK-1, were divided into ambient pressure (AP), continuous negative-pressure (NPc), and intermittent negative-pressure (NPi) groups and cultured for 24 h with scratch assay using our real-time monitoring system and device. Pressure inside the device, medium evaporation rate, and the residual wound area were compared across the groups. RESULTS: Pressure in the device was maintained at almost the same value as set in all groups. Medium evaporation rate was significantly higher in the NPi group than in the other two groups; however, it had negligible effect on cell culture. Residual wound area after 9 h evaluated by the scratch assay was significantly smaller in the NPc and NPi groups than in the AP group. CONCLUSION: We developed a negative-pressure cell culture device that enables negative-pressure cell culture under conditions similar to those used in clinical practice and is able to monitor cells under NPWT. Further experiments using this device would provide high-quality molecular biological evidence for NPWT.
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Terapia de Presión Negativa para Heridas , Humanos , Terapia de Presión Negativa para Heridas/métodos , Cicatrización de Heridas , Queratinocitos , Técnicas de Cultivo de Célula , VendajesRESUMEN
OBJECTIVES: To verify the utility of triangular extension of a hinge flap in buccal mucosal staged urethroplasty to resolve stomal stenosis after the first stage and ultimately prevent restenosis. METHODS: A total of 23 patients (triangular extension group) were studied in 2013-2019. In the first stage, buccal mucosa was transplanted, and an extended triangle portion of the mucosa was placed beside the proximal and/or distal stoma that was created when the stricture segment of the urethra was resected. In the second stage, during tubularization of the urethral plate, an incision was made at the stoma to increase the caliber to which the triangular extension was inserted. The procedure was considered successful when a 17-Fr flexible cystoscope passed through the reconstructed urethra at 6 months after the second-stage urethroplasty and no additional surgery or bougie dilation required. The clinical course of the triangular extension group was compared with 24 patients who underwent conventional staged urethroplasty (control group). RESULTS: In total, 20 patients from each group underwent second-stage surgery. No patients in the triangular extension group required additional revision surgery because of stomal stenosis after first-stage surgery, whereas five (20%) control patients did. Urethroplasty was successful in 19 patients (95%) in the triangular extension group and in 19 patients (95%) in the control group. Uroflowmetry after the second-stage surgery indicated that the mean maximum urinary flow rate was 21.5 and 15.8 mL/s after triangular extension and the control procedure, respectively (P = 0.027). CONCLUSIONS: The triangular extension technique reduces the need for revision surgery and prevents postoperative restenosis.
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Uretra , Estrechez Uretral , Constricción Patológica/etiología , Constricción Patológica/prevención & control , Constricción Patológica/cirugía , Humanos , Masculino , Mucosa Bucal/cirugía , Resultado del Tratamiento , Uretra/cirugía , Estrechez Uretral/etiología , Estrechez Uretral/prevención & control , Estrechez Uretral/cirugía , Procedimientos Quirúrgicos Urológicos Masculinos/efectos adversosRESUMEN
Carotid artery stenosis is elicited by atherosclerosis and is the main cause of cerebral thrombosis. Flow-mediated endothelial vasodilation (FMD) can be measured noninvasively to assess vascular endothelial function related to atherosclerosis. The pulse wave velocity (PWV) is used to evaluate the vascular media involved in atherosclerosis. We investigated the relationship between these measurements in 75 consecutive patients with atherosclerotic cerebral thrombosis. They were assigned to three equal groups based on the severity of carotid artery stenosis on ultrasonograms. Group 1 had no stenosis, group 2 manifested moderate stenosis (<60%), and group 3 presented with severe stenosis (≥60%). We compared the FMD and PWV among the three groups. The PWV was significantly lower in group 1 than the other two groups. The FMD was significantly lower in group 3; it was significantly lower in group 2 than group 1. There was an inverse correlation between the FMD and the severity of carotid artery stenosis. Our findings show that for assessing the severity of carotid artery stenosis, the FMD is more useful than the PWV.
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Aterosclerosis/fisiopatología , Estenosis Carotídea/fisiopatología , Trombosis Intracraneal/fisiopatología , Análisis de la Onda del Pulso , Vasodilatación/fisiología , Anciano , Aterosclerosis/complicaciones , Estenosis Carotídea/complicaciones , Estenosis Carotídea/diagnóstico , Estudios de Casos y Controles , Femenino , Humanos , Trombosis Intracraneal/complicaciones , Masculino , Persona de Mediana Edad , Índice de Severidad de la EnfermedadRESUMEN
AIM: To investigate the role of macrophage colony-stimulating factor (M-CSF) in patients with hepatocellular carcinoma (HCC) after surgery. METHODS: Expression of M-CSF, distribution of M2 macrophages (MΦs), and angiogenesis were assessed in the liver, including tumors and peritumoral liver tissues. The prognostic power of these factors was assessed. Mouse isolated hepatic MΦs or monocytes were cultured with media containing M-CSF. The concentration of vascular endothelial growth factor (VEGF) in media was assessed. Furthermore, the role of the M-CSF-matured hepatic MΦs on proliferation of the vascular endothelial cell (VEC) was investigated. RESULTS: A strong correlation between the expressions of M-CSF and CD163 was observed in the peritumoral area. Also, groups with high density of M-CSF, CD163 or CD31 showed a significantly shorter time to recurrence (TTR) than low density groups. Multivariate analysis revealed the expression of M-CSF or hepatic M2MΦs in the peritumoral area as the most crucial factor responsible for shorter TTR. Moreover, the expression of M-CSF and hepatic M2MΦs in the peritumoral area had better predictable power of overall survival. Values of VEGF in culture media were significantly greater in the hepatic MΦs compared with the monocytes. Proliferation of the VEC was greatest in the cells co-cultured with hepatic MΦs when M-CSF was present in media. CONCLUSION: M-CSF increases hepatocarcinogenesis, most likely by enhancing an angiogenic factor derived from hepatic MΦ and could be a useful target for therapy against HCC.
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Carcinoma Hepatocelular/diagnóstico , Neoplasias Hepáticas/diagnóstico , Factor Estimulante de Colonias de Macrófagos/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Animales , Antígenos CD/metabolismo , Antígenos de Diferenciación Mielomonocítica/metabolismo , Biomarcadores de Tumor/metabolismo , Carcinoma Hepatocelular/metabolismo , Proliferación Celular , Supervivencia sin Enfermedad , Células Endoteliales/citología , Femenino , Humanos , Hígado/metabolismo , Neoplasias Hepáticas/metabolismo , Macrófagos/metabolismo , Masculino , Ratones , Persona de Mediana Edad , Análisis Multivariante , Recurrencia Local de Neoplasia , Neovascularización Patológica/patología , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , Valor Predictivo de las Pruebas , Pronóstico , Receptores de Superficie Celular/metabolismo , Estudios Retrospectivos , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Hepatocellular carcinoma (HCC) is one of the most prevalent human cancers, with a rising incidence worldwide. The molecular mechanisms associated with the development of HCC are complex and include multiple interconnected molecular alterations with mounting evidence indicating an important role of microRNAs (miRNAs) in the pathogenesis of HCC. In humans, the development of HCC is commonly associated with liver cirrhosis. To study fibrosis-associated liver carcinogenesis, we used a mouse model designed to emulate the development of HCC in cirrhotic liver. Specifically, we were interested in evaluating the role of miRNAs in the molecular pathogenesis of liver carcinogenesis in male B6C3F1/J mice treated with N-nitrosodiethylamine (DEN) or carbon tetrachloride (CCl4 ) alone or a combination of DEN and CCl4 and characterized by a differential tumor incidence that increased in the following order: DENAsunto(s)
Regulación Neoplásica de la Expresión Génica
, Cirrosis Hepática Experimental/genética
, Neoplasias Hepáticas Experimentales/patología
, MicroARNs/genética
, Animales
, Tetracloruro de Carbono/toxicidad
, Línea Celular Tumoral
, Dietilnitrosamina/toxicidad
, Perfilación de la Expresión Génica
, Regulación Neoplásica de la Expresión Génica/efectos de los fármacos
, Humanos
, Cirrosis Hepática Experimental/inducido químicamente
, Neoplasias Hepáticas Experimentales/inducido químicamente
, Neoplasias Hepáticas Experimentales/genética
, Masculino
, Ratones
RESUMEN
Although a superficial circumflex iliac artery perforator (SCIP) flap has recently been widely used owing to its various advantages, reports on its use in the pediatric population are limited. A case of a supermicrosurgical reconstruction of a soft tissue defect of the ankle associated with the congenital deficiency of the tibia using a free sensate SCIP flap in a 1-year-old child has been presented. The correction of the valgus deformity of the ankle resulted in a soft tissue defect, which required flap coverage. The lateral cutaneous branch of the intercostal nerve of the flap was coapted with the deep peroneal nerve for sensory recovery. Postoperative course was uneventful and the flap completely survived. The patient was able to ambulate independently at 7 months after surgery. To the best of our knowledge, this is the youngest case of a SCIP flap transfer in literature. This case showed that young age is not a contraindication for SCIP flap transfer. It is believed that the SCIP flap procedure may be a useful option for free flap reconstruction in children.
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Tobillo/cirugía , Colgajos Tisulares Libres/trasplante , Arteria Ilíaca/cirugía , Microcirugia/métodos , Colgajo Perforante/trasplante , Procedimientos de Cirugía Plástica/métodos , Tobillo/irrigación sanguínea , Colgajos Tisulares Libres/irrigación sanguínea , Humanos , Lactante , Masculino , Colgajo Perforante/irrigación sanguíneaRESUMEN
AIM: The specific purpose of this study was to investigate the role of macrophage colony-stimulating factor (M-CSF) in initiation and progression of hepatocellular carcinoma using M-CSF-deficient mice. METHODS: M-CSF-deficient (osteopetrotic: op/op) and their littermate (LM) mice were i.p. injected with diethylnitrosamine (DEN) to induce hepatocellular carcinoma. Twenty-eight weeks after DEN administration, the tumor incidence rate and serum M-CSF levels were assessed. Furthermore, distribution of the activated macrophages and the mRNA expression of CD163 and CD204 were evaluated. Moreover, angiogenesis was analyzed in tumors. In another set of experiments, apoptosis and proliferation of the hepatocytes were examined in the acute phase after DEN administration. Isolated hepatic macrophages were cultured with or without M-CSF, and vascular endothelial growth factor (VEGF) production was assessed by enzyme-linked immunoassay. RESULTS: Tumor incidence was significantly reduced in the op/op compared with the LM mice. Serum M-CSF levels were increased in the carcinogenesis models of the LM mice. Hepatic macrophages were found only in tumors in the op/op but in both normal liver tissue and tumors in the LM mice. In the op/op group, the mRNA expression of inflammatory cytokines was significantly lower compared with the LM mice. Furthermore, apoptosis was significantly increased in the op/op than the LM mice. Angiogenesis increased in liver tumors from the LM compared with the op/op mice. Production of VEGF was greater in the hepatic macrophages incubated with M-CSF compared with those without M-CSF. CONCLUSION: Thus, M-CSF is involved in the progression of chemically induced hepatocarcinogenesis.
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BACKGROUND: It was recently reported that serum interleukin (IL)-17 levels increased in liver fibrosis associated with human alcoholic liver disease. However, the role of IL-17 in liver fibrosis has not yet been elucidated. Therefore, the aim of this study was to evaluate the role of IL-17 on cholestatic liver fibrosis. MATERIALS AND METHODS: IL-17A knockout (KO) and wild-type (WT) mice were subjected to bile duct ligation. Animals were sacrificed at designated times, and serum and liver tissues were collected. The mRNA expression of hepatic fibrotic markers was assessed, and distribution of activated hepatic stellate cells (HSCs) was determined by immunohistochemical staining. In an in vitro study, Kupffer cells (KCs) and HSCs were isolated from WT mice. KCs were cultured with IL-17A or IL-17F, and production of tumor necrosis factor α (TNF-α) and transforming growth factor ß1 (TGF-ß1) was measured. HSCs were cultured with IL-17A or IL-17F, and morphologic changes were assessed by immunohistochemical staining. RESULTS: Liver damage observed in the WT mice was significantly improved in the KO mice. Serum TNF-α and TGF-ß1 levels were significantly decreased in the KO compared with the WT mice. The hepatic mRNA expression of TNF-α, TGF-ß1, and collagen 1α1, which increased in the WT mice, also significantly decreased in the KO mice. Increased hepatic fibrosis in the WT mice was significantly improved in the KO mice. Cytokine production was increased in IL-17A-treated KCs. The most remarkable myofibroblast-like changes were observed in isolated HSCs in the presence of IL-17A. CONCLUSIONS: IL-17A was involved in the pathogenesis of cholestatic liver fibrosis by activation of both the KCs and HSCs.
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Células Estrelladas Hepáticas/patología , Interleucina-17/fisiología , Macrófagos del Hígado/patología , Cirrosis Hepática/patología , Cirrosis Hepática/fisiopatología , Actinas/metabolismo , Animales , Células Cultivadas , Colágeno Tipo I/metabolismo , Modelos Animales de Enfermedad , Células Estrelladas Hepáticas/efectos de los fármacos , Células Estrelladas Hepáticas/metabolismo , Hidroxiprolina/metabolismo , Técnicas In Vitro , Interleucina-17/genética , Interleucina-17/farmacología , Macrófagos del Hígado/efectos de los fármacos , Macrófagos del Hígado/metabolismo , Cirrosis Hepática/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Factor de Crecimiento Transformador beta1/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
BACKGROUND: Liver regeneration after partial hepatectomy (PH) is regulated by tumor necrosis factor (TNF)-α derived from the Kupffer cell. Furthermore, it was reported from our laboratory that interleukin (IL)-17A enhances the production of TNF-α by the Kupffer cell, suggesting that IL-17A may play a role in liver regeneration. OBJECTIVE: The purpose was to determine the role of IL-17A and the spleen in liver regeneration after PH. METHODS: Two mouse models including the wild-type (WT) mice or the IL-17A knockout (KO) mice underwent PH. Animals were killed at the designated time points; liver tissues were harvested for further investigation. Proliferation of hepatocytes was evaluated. Furthermore, the messenger RNA and protein expression of TNF-α and IL-6 were measured in the liver. In another set of experiments, the two animal models underwent splenectomy before PH. In an in vitro study, CD4-positive lymphocytes in the spleen were isolated from mice, and the number of IL-17A-positive cells was investigated. RESULTS: Liver regeneration was significantly impaired in the KO mice compared with the WT mice. This was associated with suppression of cell proliferation assessed by cell proliferation markers in the KO mice. In the WT mice that underwent splenectomy, liver regeneration was significantly delayed compared with animals without splenectomy. In contrast, splenectomy did not affect liver regeneration in the KO mice. IL-17A-positive lymphocytes increased significantly in the spleen in the WT mice after PH. CONCLUSIONS: These results indicate that IL-17A derived from CD4-positive lymphocytes in the spleen is a key regulator in liver regeneration after PH.
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Hepatectomía/métodos , Interleucina-17/fisiología , Regeneración Hepática/fisiología , Hígado/fisiología , Hígado/cirugía , Animales , Linfocitos T CD4-Positivos/patología , Proliferación Celular , Hepatocitos/patología , Técnicas In Vitro , Interleucina-17/deficiencia , Interleucina-17/genética , Interleucina-6/fisiología , Hígado/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Modelos Animales , Bazo/patología , Bazo/fisiología , Bazo/cirugía , Esplenectomía , Factor de Necrosis Tumoral alfa/fisiologíaRESUMEN
UNLABELLED: Alcoholic liver injury is a major public health issue worldwide. Even though the major mechanisms of this disease have been established over the past decades, little is known about genetic susceptibility factors that may predispose individuals who abuse alcoholic beverages to liver damage and subsequent pathological conditions. We hypothesized that a panel of genetically diverse mouse strains may be used to examine the role of endoplasmic reticulum (ER) stress and one-carbon metabolism in the mechanism of interindividual variability in alcoholic liver injury. We administered alcohol (up to 27 mg/kg/d) in a high-fat diet using an intragastric intubation model for 28 days to male mice from 14 inbred strains (129S1/SvImJ, AKR/J, BALB/cJ, BALB/cByJ, BTBR T+tf/J, C3H/HeJ, C57BL/10J, DBA/2J, FVB/NJ, KK/HIJ, MOLF/EiJ, NZW/LacJ, PWD/PhJ, and WSB/EiJ). Profound interstrain differences (more than 3-fold) in alcohol-induced steatohepatitis were observed among the strains in spite of consistently high levels of urine alcohol that were monitored throughout the study. We found that ER stress genes were induced only in strains with the most liver injury. Liver glutathione and methyl donor levels were affected in all strains, albeit to a different degree. The most pronounced effects that were closely associated with the degree of liver injury were hyperhomocysteinemia and strain-dependent differences in expression patterns of one-carbon metabolism-related genes. CONCLUSION: Our data demonstrate that strain differences in alcohol-induced liver injury and steatosis are striking and independent of alcohol exposure and the most severely affected strains exhibit major differences in the expression of ER stress markers and genes of one-carbon metabolism.
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Alcoholes/administración & dosificación , Hígado Graso Alcohólico/metabolismo , Hígado Graso Alcohólico/patología , Ratones Endogámicos/metabolismo , Alcoholes/efectos adversos , Animales , Biopsia con Aguja , Western Blotting , Modelos Animales de Enfermedad , Inmunohistoquímica , Peroxidación de Lípido/fisiología , Masculino , Metionina/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Transferasas del Grupo 1-Carbono/metabolismo , Estrés Oxidativo/fisiología , Distribución Aleatoria , Índice de Severidad de la Enfermedad , Especificidad de la EspecieRESUMEN
OBJECTIVE: The purpose of this study was to investigate whether inhibition of Kupffer cells (KCs) affects the expression of high mobility group box 1 (HMGB1) and mortality in septic peritonitis. The role of the spleen in septic peritonitis was also investigated. METHODS: Rats were given liposome-entrapped dichloromethylene diphosphonate (lipo-MDP) to eliminate KCs or non-entrapped liposome (lipo) before cecal ligation and puncture (CLP), and serum HMGB1 levels and mortality were assessed after CLP. Furthermore, KCs and tissue macrophages were isolated, and production of HMGB1 was investigated. Effects of splenectomy on serum HMGB1 levels and mortality were also investigated after CLP. RESULTS: Elimination of the Kupffer cells by lipo-MDP increased serum HMGB1 concentrations and mortality significantly. Furthermore, HMGB1 expression in both the periportal area of the liver and the spleen was greater in the lipo-MDP group than the lipo group. On the other hand, splenectomy blunted serum HMGB1 levels and improved mortality after CLP. The HMGB1 expression was greater in the spleen compared with the liver after CLP. Furthermore, production of HMGB1 was greatest in splenic macrophages in vitro. The number of ED3-positive cells increased significantly in non-splenectomized animals but not in splenectomized animals after CLP. In the lipo-MDP treated groups, the number of ED3-positive macrophages also increased in the liver from non-splenectomized animals but not in the splenectomized animals after CLP. CONCLUSIONS: The liver and the spleen play key roles in host defense during septic peritonitis. Migrating macrophages into the liver are, in part, derived from the spleen after CLP.
Asunto(s)
Proteína HMGB1/biosíntesis , Macrófagos del Hígado/metabolismo , Peritonitis/mortalidad , Sepsis/fisiopatología , Esplenectomía , Animales , Modelos Animales de Enfermedad , Masculino , Peritonitis/metabolismo , Ratas , Ratas Sprague-Dawley , Sepsis/mortalidadRESUMEN
BACKGROUND: Interleukin (IL)-17A is a proinflammatory cytokine and plays an important role in neutrophil recruitment. We investigate the role of IL-17A in a mouse polymicrobial sepsis model. MATERIALS AND METHODS: IL-17A knockout mice (KO) and wild-type (WT) mice were subjected the cecal ligation and puncture (CLP). Survival was assessed for the following 7 d after the CLP operation, and histopathologic findings were evaluated 12 h after CLP. Bacterial outgrowth in blood was assessed by blood culture 12 h after CLP. After CLP, expression of inflammatory mediators in serum was assessed by enzyme-linked immunosorbent assay (ELISA). Furthermore, expression of FOXP3 and IL-17A in the spleen was assessed by immunohistochemical staining and flow cytometry. RESULTS: Mortality was increased in KO mice compared with WT mice after CLP. Furthermore, bacterial outgrowth in blood and serum high mobility group box 1 (HMGB1) levels were also significantly greater in KO mice than WT mice. The expression of FOXP3 in the spleen was significantly greater in KO mice than WT mice. CONCLUSION: IL-17A play pivotal role in host defense during septic peritonitis.
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Interleucina-17/fisiología , Sepsis/etiología , Alanina Transaminasa/sangre , Animales , Traslocación Bacteriana , Citocinas/sangre , Femenino , Factores de Transcripción Forkhead/análisis , Proteína HMGB1/análisis , Interleucina-17/análisis , Pulmón/enzimología , Pulmón/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Peroxidasa/metabolismo , Sepsis/mortalidad , Bazo/químicaRESUMEN
Recent evidence suggests that IL-17A regulates neutrophil-dependent organ injury. Accordingly, the purpose of this study was to determine the role of IL-17A in neutrophil recruitment after ischemia-reperfusion (I/R) and in subsequent liver injury. Two mouse models including wild-type and IL-17A knockout mice were evaluated for I/R injury. The medial largest lobe of the liver was clamped for 90 min. In another set of experiments, recombinant mouse (rm)IL-17A homodimer or rmIL-17A/F heterodimer were administered to knockout mice before I/R, and liver injury was investigated. Isolated Kupffer cells were incubated with rmIL-17A or rmIL-17F, and production of TNF-α was measured. Studies evaluating the extent of liver injury as measured by serum transaminase levels demonstrated similar levels in the acute phase (6 h) in these two models. In contrast, in the subacute phase (20 h) after I/R, both serum transaminase levels and percent of hepatic necrosis were significantly reduced in the knockout mice compared with the wild-type mice. This reduction in liver injury seen in the knockout mice was associated with suppression of chemokine and adhesion molecule expression and reduction in infiltration of neutrophils into the liver. Administration of rmIL-17A homodimer, but not IL-17A/F heterodimer, increased liver injury in the subacute phase of I/R in KO mice. TNF-α production by isolated Kupffer cells increased significantly in the cells incubated with rmIL-17A compared with rmIL-17F. These results indicate that IL-17A is a key regulator in initiating neutrophil-induced inflammatory responses and hepatic injury in the subacute phase after reperfusion.
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Interleucina-17/fisiología , Hígado/irrigación sanguínea , Infiltración Neutrófila/inmunología , Neutrófilos/inmunología , Daño por Reperfusión/inmunología , Daño por Reperfusión/metabolismo , Animales , Temperatura Corporal/genética , Temperatura Corporal/inmunología , Modelos Animales de Enfermedad , Interleucina-17/deficiencia , Interleucina-17/genética , Macrófagos del Hígado/inmunología , Macrófagos del Hígado/patología , Hígado/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Necrosis , Infiltración Neutrófila/genética , Neutrófilos/patología , Daño por Reperfusión/patología , Células Th17/inmunología , Células Th17/metabolismo , Células Th17/patologíaRESUMEN
High-mobility group box 1 (HMGB1) acts as an early mediator of inflammation and organ damage in hepatic ischemia-reperfusion (I/R) injury. Glycyrrhizin is a natural anti-inflammatory and antiviral triterpene in clinical use. The purpose of this study was to investigate the effect of glycyrrhizin on liver injury caused by I/R and production of HMGB1 by Kupffer cells in rats. In the first test period, rats were given saline or glycyrrhizin 20 min before segmental hepatic warm I/R. Serum alanine aminotransferase and HMGB1 levels and hepatic histopathological findings were evaluated after I/R. Furthermore, expression of HMGB1 in the liver was assessed by immunohistochemical staining after I/R. Kupffer cells were isolated by collagenase digestion and differential centrifugation, and production of HMGB1 was assessed. In another set of experiments, the effect of inhibition of Kupffer cells by injection of liposome-entrapped dichloromethylene diphosphonate (lipo-MDP) on liver injury and expression of HMGB1 were investigated after I/R. Liver injury was prevented in the glycyrrhizin group compared with the control group. Furthermore, serum HMGB1 levels were also significantly blunted in the glycyrrhizin group compared with the control group. Cells expressing HMGB1 were detected in the hepatic sinusoid by immunohistochemistry and recognized morphologically as Kupffer cells. Furthermore, the expression of HMGB1 was reduced in the glycyrrhizin group compared with the control group. Production of HMGB1 was reduced in Kupffer cells isolated from the glycyrrhizin group compared with the control group. It is noteworthy that treatment with lipo-MDP significantly blunted serum HMGB1 levels and prevented liver injury after I/R. These results suggest that glycyrrhizin has the therapeutic potential to prevent warm I/R-induced injury during hepato-biliary surgery.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Ácido Glicirrínico/farmacología , Proteína HMGB1/antagonistas & inhibidores , Proteína HMGB1/biosíntesis , Macrófagos del Hígado/metabolismo , Daño por Reperfusión/patología , Alanina Transaminasa/metabolismo , Animales , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , Ácido Clodrónico/química , Ácido Clodrónico/farmacología , Portadores de Fármacos , Inmunohistoquímica , Macrófagos del Hígado/efectos de los fármacos , Liposomas , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
PURPOSE: It was reported from this laboratory that Kupffer cells (KCs) were activated in patients infected with HCV. Since dendritic cells, monocytes, and macrophages were activated by stimulation with HCV-related proteins, the specific aim of this study was to investigate the role of HCV-related proteins in activation of KCs, the signal pathway of activation of KCs mediated by Toll-like receptor (TLR) 4, and the influence of HCV infection on function of KCs. METHODS: Kupffer cells isolated from non-cancerous surgical specimen were co-cultured with HCV-related proteins (Core, NS3, NS4, and NS5), and production of cytokines (TNF-α, IL-1ß, and IL-10) and hydrogen peroxide were assessed. Furthermore, effects of neutralization antibodies against the TLR2, TLR3, or TLR4, and cytochalasin B on the production TNF-α by KCs were investigated. RESULTS: Kupffer cells produced markedly a proinflammatory cytokine TNF-α by stimulation with all HCV-related proteins studied, and values were as same as production by KCs stimulated with LPS. Importantly, this production in the case of NS3 was significantly blunted by about 60% by neutralization antibodies against the TLR4, but not cytochalasin B. Production of TNF-α by isolated KCs stimulated with LPS was significantly greater in the HCV-infected livers than the HCV/HBV-negative livers. CONCLUSIONS: These results indicated that HCV-related proteins may cause prolonged activation of KCs in the HCV-infected liver, leading to accumulation of inflammatory cytokines that contribute to DNA damage and carcinogenesis. Furthermore, function of KCs was difference between patients infected with and without HCV infection.
Asunto(s)
Hepacivirus/metabolismo , Hepatitis C/metabolismo , Macrófagos del Hígado/virología , Hígado/virología , Proteínas Virales/metabolismo , Anciano , Anticuerpos Neutralizantes/metabolismo , Células Cultivadas , Técnicas de Cocultivo , Citocalasina B/metabolismo , Femenino , Humanos , Peróxido de Hidrógeno/metabolismo , Interleucina-10/biosíntesis , Interleucina-1beta/biosíntesis , Macrófagos del Hígado/efectos de los fármacos , Macrófagos del Hígado/metabolismo , Lipopolisacáridos/farmacología , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Persona de Mediana Edad , Receptores Toll-Like/metabolismo , Factor de Necrosis Tumoral alfa/biosíntesis , Proteínas Virales/farmacologíaRESUMEN
The specific purpose of this study was to evaluate the significant effects of medium-chain triglycerides (MCTs) and N-3 fatty acids on chemically induced experimental colitis induced by 2,4,6-trinitrobenzene sulphonic acid (TNBS) in rats. Male Wistar rats were fed liquid diets enriched with N-6 fatty acid (control diets), N-3 fatty acid (MCT- diets), and N-3 fatty acid and MCT (MCT+ diets) for 2 weeks and then were given an intracolonic injection of TNBS. Serum and tissue samples were collected 5 days after ethanol or TNBS enema. The severity of colitis was evaluated pathologically, and tissue myeloperoxidase activity was measured in colonic tissues. Furthermore, protein levels for inflammatory cytokines and a chemokine were assessed by an enzyme-linked immunosorbent assay in colonic tissues. Induction of proinflammatory cytokines tumor necrosis factor-α and interleukin-1ß in the colon by TNBS enema was markedly attenuated by the MCT+ diet among the 3 diets studied. Furthermore, the induction of chemokines macrophage inflammatory protein-2 and monocyte chemotactic protein-1 also was blunted significantly in animals fed the MCT+ diets. As a result, MPO activities in the colonic tissue also were blunted significantly in animals fed the MCT+ diets compared with those fed the control diets or the MCT- diets. Furthermore, the MCT+ diet improved chemically induced colitis significantly among the 3 diets studied. Diets enriched with both MCTs and N-3 fatty acids may be effective for the therapy of inflammatory bowel disease as antiinflammatory immunomodulating nutrients.
Asunto(s)
Colitis/prevención & control , Nutrición Enteral/métodos , Ácidos Grasos Omega-3/administración & dosificación , Triglicéridos/administración & dosificación , Alimentación Animal , Animales , Peso Corporal/efectos de los fármacos , Quimiocinas/metabolismo , Colitis/inducido químicamente , Colitis/patología , Colon/efectos de los fármacos , Colon/enzimología , Colon/patología , Modelos Animales de Enfermedad , Endotoxemia/sangre , Endotoxemia/tratamiento farmacológico , Endotoxemia/etiología , Endotoxinas/sangre , Enema , Masculino , Peroxidasa/metabolismo , Ratas , Ratas Wistar , Ácido Trinitrobencenosulfónico/toxicidadRESUMEN
Transcriptional silencing of tumor suppressor genes and other cancer-related genes induced by promoter CpG island hypermethylation is an important epigenetic mechanism of hepatocarcinogenesis. Previous studies have established methylation profiles of hepatocellular carcinomas (HCCs) and demonstrated that methylation of several candidate genes in resected tissues may be associated with time to recurrence. The goals of our study were to test whether specific promoter methylation and mRNA levels of candidate genes, as well as global changes in DNA methylation, can be linked with time to recurrence and clinicopathological variables in a homogenous study group of HCC patients. Forty-three tumorous and 45 non-tumorous liver tissue samples from the surgical margin were obtained from HCV-positive, HBV-negative HCC patients who underwent tumor resection surgery and who were monitored for tumor recurrence thereafter (median follow-up time: 16 months (range, 0-79 months)). Methylation-specific PCR was used to assess the promoter methylation status of P16(INK4a), SOCS-1, RASSF1A, APC, GSTP1, RIZ1, and MGMT genes, while the level of LINE-1 methylation was used as marker of global DNA methylation levels. Methylation frequencies in P16(INK4a), RASSF1A, APC, GSTP1, and RIZ1 genes were significantly greater in tumorous versus non-tumorous tissues. Methylation of RIZ1 in non-tumorous tissues was significantly associated with time to recurrence. Additionally, genomic DNA was significantly more hypomethylated in tumorous tissues, and this change was associated with shorter recurrence, but not with clinicopathological features. In conclusion, this study supports the role of aberrant methylation in the pathobiology of HCV-positive HCCs. The finding that RIZ1 methylation and increased levels of LINE-1 hypomethylation in non-tumorous tissues are associated with time to recurrence underscores the importance of assessing the epigenetic state of the liver remnant.