ABSTRACT
Abstract Fluorescent nanodiamond (FND) has been used for long-term cell labeling and in vivo cell tracking because they have good at photostability and biocompatibility. In this study, we evaluate the effect of fluorescent nanodiamond labeling on in vitro culture and differentiation of human umbilical cord mesenchymal stem cells (hUCMSCs) into hepatocyte-like cells (HLCs). For hepatic differentiation of hUCMSCs, cells were induced with human hepatocyte growth factor, nicotinamide and Dexamethasone. FND was supplied in two experimental groups with 20 μg/mL and 100 μg/mL in 2 hours. The cell was assessed for FND uptake by laser scan microscopy and flow cytometry methods. The effect of FND on hUCMSCs was evaluated by the cell viability and growth assays as well as the differentiation throughout of morphology alterations or gene expression of anfa-fetoprotein, albumin, and hepatocyte nuclear factor 4α. The results showed that the labeling of hUCMSCs is efficient and easy and there was significant cellular uptake of FND. We did not observe any negative impacts of FND to the cell viability and growth. FND can be utilized for the long-term labeling and tracking of hUCSCs and HLCs in vivo studies.
Subject(s)
Humans , Umbilical Cord/cytology , Cell Differentiation , Hepatocytes/cytology , Mesenchymal Stem Cells/cytology , Cell Survival , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Abstract Diplopods are considered important macroarthropods the soil as part of its maintenance and balance. These animals usually do not occur in high densities, but population explosions caused by environmental disturbances, climate changes, and use of pesticides that eliminate possible competitors, have been reported. The millipede Urostreptus atrobrunneus Pierozzi and Fontanetti, 2006 have become a nuisance to humans in infestation sites in urban centers of the state of Sao Paulo, Brazil. As a contribution to the understanding of this potential pest, this study describes the histology, histochemistry, and ultrastructure of the U. atrobrunneus midgut, and presents the redefinition of hepatic cells somewhat controversial in the literature. The region of the midgut is characterized by the absence of a cuticular intima, and composed of a pseudostratified epithelium on a thick basal membrane, followed by a muscle layer, a layer of hepatic cells, lined by an external membrane. The morphology observed in U. atrobrunneus is similar to that reported for other species of diplopods. The hepatic cells have been previously described as randomly without forming a layer, however, the present results clearly demonstrate that these cells form a continuous layer over the whole midgut.
Resumo Diplópodos são considerados importantes macro-artrópodes do solo, uma vez que participam de sua manutenção e equilíbrio. Comumente estes animais não apresentam população numerosa, porém há relatos de explosões populacionais ocasionadas por desequilíbrios ambientais, mudanças climáticas e utilização de pesticidas que eliminam possíveis competidores. O milípede Urostreptus atrobrunneus Pierozzi e Fontanetti, 2006, têm apresentado pontos de infestação em centros urbanos, do estado de São Paulo, Brasil, causando muitos transtornos à população humana. Com objetivo de contribuir para o conhecimento desta potencial praga, este trabalho apresenta a descrição histológica, histoquímica e ultra-estrutural do intestino médio do milípede U. atrobrunneus, bem como apresenta a redefinição das células hepáticas, um tanto controversa na literatura pertinente. A região do intestino médio é caracterizada pela ausência da íntima cuticular, sendo formado por um epitélio pseudoestratificado, apoiado por uma membrana basal espessa, seguido de uma camada muscular, uma camada de células hepáticas, revestido por uma membrana externa. A morfologia observada neste trabalho assemelha-se bastante com as descrições de outras espécies de diplópodos estudadas até o momento. As células hepáticas foram previamente descritas como dispostas aleatoriamente sem a formação de uma camada, contudo, os presentes resultados demonstram claramente que estas células formam uma camada contínua ao longo de todo o intestino médio.
Subject(s)
Animals , Arthropods/anatomy & histology , Hepatocytes/cytology , Gastrointestinal Tract/anatomy & histologyABSTRACT
We describe and compare the histology of liver and spleen ofGeophagus brasiliensis (Perciformes), Hypostomus francisci (Siluriformes) and Hoplias aff. malabaricus (Characiformes), tropical freshwater fishes. InG. brasiliensisandH. aff. malabaricusthe hepatocytes were arranged in tubular form whereas in H. franciscithey cord-like. In all species, hepatocytes presented glycogen, but in G. brasiliensis and H. aff. malabaricus they showed strong stained for hemossiderin in the cytoplasm. InG. brasiliensis and H. aff. malabaricus, melanomacrophage centres (MMCs) were associated to hepatic structures and only in G. brasiliensis was observed intrahepatic exocrine pancreas. The spleen, in all species, was characterized by red and white pulp without boundary between the two regions, but only in H. francisci was recorded nodular organization in splenic parenchyma. The G. brasiliensisandH. aff. malabaricuspresented in the white pulp MMCs linked mainly to ellipsoids. Besides, we observed large MMCs in the spleen in relation to liver of G. brasiliensis and H. aff. malabaricus. In liver, highest values of reticular fibers and collagen were observed inG. brasiliensis. In spleen, highest values of reticular fibers and collagen were recorded inH. aff. malabaricusandH. francisci, respectively. Histological differences confirm the hypothesis that the phylogenetic distance is reflected in liver and spleen.(AU)
Nós descrevemos e comparamos a histologia do fígado e do baço de Geophagus brasiliensis (Perciformes), Hypostomus francisci (Siluriformes) e Hoplias aff. malabaricus (Characiformes), peixes neotropicais de água doce. Em G. brasiliensis e H. aff. malabaricus os hepatócitos organizaram-se na forma tubular enquanto que em H. francisci eles apresentaram-se como cordões celulares. Em todas as espécies, os hepatócitos apresentaram glicogênio, mas em G. brasiliensis e H. aff. malabaricus, eles mostraram forte marcação para hemossiderina no citoplasma. Em G. brasiliensis e H. aff. malabaricus, centros melanomacrofágicos (CMMs) foram associados a estruturas hepáticas e somente em G. brasiliensis foi observado pâncreas exócrino intrahepático. O baço, em todas as espécies, foi caracterizado pela polpa vermelha e branca sem limites entre as duas regiões, mas somente em H. francisci foi registrado uma organização nodular no parênquima esplênico. G. brasiliensiseH. aff. malabaricusapresentaram na polpa branca CMMs associados principalmente a elipsoides. Além disso, nós observamos CMMs grandes no baço em relação ao fígado de G. brasiliensis e de H. aff. malabaricus. No fígado, valores altos de fibras reticulares e colágeno foram observado em G. brasiliensis. No baço, valores altos de fibras reticulares e colágeno foram registrados em H. aff. malabaricuseH. francisci, respectivamente. Diferenças histológicas confirmam a hipótese que a distância filogenética está refletida no fígado e no baço.(AU)
Subject(s)
Animals , Fishes/abnormalities , Fishes/anatomy & histology , Hepatocytes/cytology , Liver/anatomy & histology , Spleen/anatomy & histologyABSTRACT
BACKGROUND/AIMS: Chronic liver disease leads to liver fibrosis, and although the liver does have a certain regenerative capacity, this disease is associated with dysfunction of the liver vessels. C-reactive protein (CRP) is produced in the liver and circulated from there for metabolism. CRP was recently shown to inhibit angiogenesis by inducing endothelial cell dysfunction. The objective of this study was to determine the effect of CRP levels on angiogenesis in a rat model of liver dysfunction induced by bile duct ligation (BDL). METHODS: The diameter of the hepatic vein was analyzed in rat liver tissues using hematoxylin and eosin (H&E) staining. The expression levels of angiogenic factors, albumin, and CRP were analyzed by real-time PCR and Western blotting. A tube formation assay was performed to confirm the effect of CRP on angiogenesis in human umbilical vein endothelial cells (HUVECs) treated with lithocholic acid (LCA) and siRNA-CRP. RESULTS: The diameter of the hepatic portal vein increased significantly with the progression of cirrhosis. The expression levels of angiogenic factors were increased in the cirrhotic liver. In contrast, the expression levels of albumin and CRP were significantly lower in the liver tissue obtained from the BDL rat model than in the normal liver. The CRP level was correlated with the expression of albumin in hepatocytes treated with LCA and siRNA-CRP. Tube formation was significantly decreased in HUVECs when they were treated with LCA or a combination of LCA and siRNA-CRP. CONCLUSION: CRP seems to be involved in the abnormal formation of vessels in hepatic disease, and so it could be a useful diagnostic marker for hepatic disease.
Subject(s)
Animals , Humans , Male , Rats , Angiogenic Proteins/genetics , Bile Ducts/surgery , C-Reactive Protein/analysis , Cells, Cultured , Disease Models, Animal , Hepatic Veins/abnormalities , Hepatocytes/cytology , Human Umbilical Vein Endothelial Cells , Lithocholic Acid/pharmacology , Liver/metabolism , Liver Cirrhosis/etiology , Liver Diseases/metabolism , Microscopy, Fluorescence , Mitochondria/drug effects , RNA Interference , RNA, Small Interfering/metabolism , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Serum Albumin/geneticsABSTRACT
Gap junctions mediate direct communication between cells; however, toxicological cascade triggered by nonessential metals can abrogate cellular signaling mediated by gap junctions. Although cadmium (Cd) is known to induce apoptosis in organs and tissues, the mechanisms that underlie gap junction activity in Cd-induced apoptosis in BRL 3A rat liver cells has yet to be established. In this study, we showed that Cd treatment decreased the cell index (a measure of cellular electrical impedance) in BRL 3A cells. Mechanistically, we found that Cd exposure decreased expression of connexin 43 (Cx43), increased expression of p-Cx43 and elevated intracellular free Ca2+ concentration, corresponding to a decrease in gap junctional intercellular communication. Gap junction blockage pretreatment with 18β-glycyrrhizic acid (GA) promoted Cd-induced apoptosis, involving changes in expression of Bax, Bcl-2, caspase-3 and the mitochondrial transmembrane electrical potential (Δψm). Additionally, GA was found to enhance ERK and p38 activation during Cd-induced activation of mitogen-activated protein kinases, but had no significant effect on JNK activation. Our results indicated the apoptosis-related proteins and the ERK and p38 signaling pathways may participate in gap junction blockage promoting Cd-induced apoptosis in BRL 3A cells.
Subject(s)
Animals , Rats , Apoptosis/drug effects , Cadmium/toxicity , Calcium/metabolism , Cell Communication/drug effects , Connexin 43/genetics , Enzyme Activation/drug effects , Gap Junctions/drug effects , Gene Expression Regulation/drug effects , Hepatocytes/cytology , Signal Transduction/drug effectsABSTRACT
Purpose To determine the safety of continued administration of antithrombotic agents during transperineal (TP) prostate biopsy. Patients and Methods A total of 811 men who underwent transrectal ultrasound (TRUS)-guided TP biopsy from January 2008 to June 2012 at our two institutions were retrospectively analyzed. Among these 811 men, 672 received no antithrombotic agents (group I), 103 received and continued administration of antithrombotic agents (group II), and 36 interrupted administration of antithrombotic agents (group III). Overall complications were graded and hemorrhagic complications were compared (group I with group II) using propensity score matching (PSM) analysis. Results An overall complication rate of 4.6% was recorded. Hemorrhagic complications occurred in 1.8% and they were virtually identical in all the three groups, and no severe hemorrhagic complications occurred. One patient in group III required intensive care unit admission for cerebral infarction. PSM analysis revealed no statistical difference between groups I and II with regard to the incidence of gross hematuria, perineal hematoma, and rectal bleeding. Multiple regression analysis revealed that hemorrhagic complications were associated with lower body mass index (<21 kg/m2, P=0.0058), but not with administration of antithrombotic agents. Conclusions Continued administration of antithrombotic agents does not increase the risk of hemorrhagic complications; these agents are well tolerated during TP biopsy. .
Subject(s)
Adolescent , Adult , Animals , Child , Child, Preschool , Female , Humans , Infant , Male , Mice , Young Adult , Hepatocytes/transplantation , Liver Diseases/pathology , Liver Transplantation/methods , Liver/pathology , Hepatocytes/cytology , Hepatocytes/metabolism , Liver Diseases/metabolism , Liver Diseases/surgery , Liver/metabolism , Liver/surgeryABSTRACT
Based on their ability to differentiate into multiple cell types including hepatocytes, the transplantation of mesenchymal stem cells (MSCs) has been suggested as an effective therapy for chronic liver diseases. The aim of this study was to evaluate the safety, efficacy and therapeutic effects of MSCs in patients with chronic liver disease through a literature-based examination. We performed a systematic review (SR) and meta-analysis (MA) of the literature using the Ovid-MEDLINE, EMBASE and Cochrane Library databases (up to November 2014) to identify clinical studies in which patients with liver diseases were treated with MSC therapy. Of the 568 studies identified by the initial literature search, we analyzed 14 studies and 448 patients based on our selection criteria. None of the studies reported the occurrence of statistically significant adverse events, side effects or complications. The majority of the analyzed studies showed improvements in liver function, ascites and encephalopathy. In particular, an MA showed that MSC therapy improved the total bilirubin level, the serum albumin level and the Model for End-stage Liver Disease (MELD) score after MSC treatment. Based on these results, MSC transplantation is considered to be safe for the treatment of chronic liver disease. However, although MSCs are potential therapeutic agents that may improve liver function, in order to obtain meaningful insights into their clinical efficacy, further robust clinical studies must be conducted to evaluate the clinical outcomes, such as histological improvement, increased survival and reduced liver-related complications, in patients with chronic liver disease.
Subject(s)
Humans , Cell Differentiation/physiology , Cell- and Tissue-Based Therapy/adverse effects , Hepatocytes/cytology , Liver/physiopathology , Liver Diseases/therapy , Liver Function Tests , Mesenchymal Stem Cell Transplantation/adverse effects , Mesenchymal Stem Cells/cytologyABSTRACT
The direct differentiation of hepatocytes from bone marrow cells remains controversial. Several mechanisms, including transdifferentiation and cell fusion, have been proposed for this phenomenon, although direct visualization of the process and the underlying mechanisms have not been reported. In this study, we established an efficient in vitro culture method for differentiation of functioning hepatocytes from murine lineage-negative bone marrow cells. These cells reduced liver damage and incorporated into hepatic parenchyma in two independent hepatic injury models. Our simple and efficient in vitro protocol for endodermal precursor cell survival and expansion enabled us to identify these cells as existing in Sca1+ subpopulations of lineage-negative bone marrow cells. The endodermal precursor cells followed a sequential developmental pathway that included endodermal cells and hepatocyte precursor cells, which indicates that lineage-negative bone marrow cells contain more diverse multipotent stem cells than considered previously. The presence of equivalent endodermal precursor populations in human bone marrow would facilitate the development of these cells into an effective treatment modality for chronic liver diseases.
Subject(s)
Animals , Female , Mice , Ataxin-1/analysis , Bone Marrow Cells/cytology , Cell Differentiation , Cell Proliferation , Cells, Cultured , Hepatocytes/cytology , Mice, Inbred BALB CABSTRACT
Embora o trabalho seja essencial à vida humana, o contexto no qual é realizado pode desencadear tensão, desajuste e consequente adoecimento. Estudos apontam a Enfermagem como uma ocupação com alto risco para adoecimento, em especial no ambiente hospitalar. O objetivo deste estudo foi estimar a ocorrência de agravos à saúde referidos pelos trabalhadores de enfermagem em um hospital na Bahia. Estudo de corte transversal envolvendo 309 profissionais. As queixas de saúde mais frequentes relacionaram-se a sintomas osteomusculares como dor nas pernas 66,4% (192) e dor nas costas 61,8% (178); e à saúde mental com predomínio de cansaço mental 47% (131) e nervosismo 33,7% (93). Os resultados apontam que enfermeiros apresentaram maior frequência de agravos posturais e queixas de saúde mental enquanto as queixas de agravos respiratórios foram mais prevalentes entre técnicos e auxiliares. Estes resultados evidenciam necessidade de maior consciência do processo de trabalho e realização de atividades preventivas.
Although the work is essential to human life, the context in which it is done can induct tension, imbalance and consequent illness. Studies indicate nursing as an occupation with high risk for disease, particularly in the hospital environment. The aim of this study was to estimate the occurrence of health problems reported by nursing staff in a hospital in Bahia, Brazil. This is a cross-sectional study, involving 309 nursing professionals. The most frequent health complaints were related to musculoskeletal symptoms such as leg pain 66.4% (192) and back pain 61.8% (178); and to mental health with a predominance of mental fatigue 47% (131) 33 and nervousness 7% (93). The results show that nurses had a higher frequency of postural problems and mental health complaints while respiratory diseases complaints were more prevalent among technicians and assistants. These results highlight the need for greater awareness of the work process and implementation of preventive activities.
Aunque el trabajo es esencial para la vida humana, el contexto en el que se realiza puede desencadenar tensión, desequilibrio y enfermedad consecuente. Los estudios muestran la enfermería como una profesión de alto riesgo para la enfermedad, sobre todo en el medio hospitalario. El objetivo de este estudio fue estimar la incidencia de problemas de salud reportados por el personal de enfermería en un hospital de Bahia, Brasil. Trata-se de un estudio transversal con 309 profesionales. Los problemas de salud más frecuentes relacionadas con los síntomas musculo esqueléticos, como el dolor de pierna 66.4% (192) y el dolor de espalda 61,8% (178) y la salud mental, con un predominio de la fatiga mental 47% (131) 33 y el nerviosismo 7% (93). Los resultados muestran que las enfermeras tenían una mayor frecuencia de problemas de postura y quejas de salud mental mientras que las quejas de las enfermedades respiratorias son más frecuentes entre los técnicos y auxiliares. Estos resultados ponen de relieve la necesidad de una mayor conciencia sobre el proceso de trabajo y la ejecución de las actividades preventivas.
Subject(s)
Animals , Rats , Drugs, Chinese Herbal/pharmacology , Hepatocytes/cytology , Hepatocytes/drug effects , Apoptosis/drug effects , Cell Line , Cell Division/drug effects , Liver Cirrhosis/drug therapy , Liver Cirrhosis/pathology , PhytotherapyABSTRACT
FUNDAMENTO: la regeneración hepática es la respuesta fundamental del hígado frente a los diversos estímulos internos y externos. Es multifactorial inducida, que origina cambios secuenciales en la expresión génica, la estructura y organización de las células hepáticas. OBJETIVOS: analizar el comportamiento de la densidad superficial de los núcleos de los hepatocitos según la posición que ocupan en las zonas del lóbulo hepático medio de ratones sometidos a citosol regenerativo. Método: se estudió la densidad superficial de los núcleos de hepatocitos con el uso de la morfometría en 14 lóbulos medios de hígados de ratones, que fueron sometidos a dosis de citosol de hígado regenerativo al inicio del experimento. Las muestras fueron tomadas cada 12 horas durante ocho días. El corte se dividió en zonas proximal, medial y distal al hilio donde se realizaron las determinaciones. RESULTADOS: los valores promedios y su desviación estándar en la zonas fueron: proximal 701,23 ± 135,83 (μm2.103), medial 707,48 ± 46,87 (μm2.103) y distal 682,48 ± 125,21 (μm2.103). Al comparar los resultados mediante el análisis de varianza se aprecia que no existen diferencias en el comportamiento de los promedios en las tres zonas. Al comparar la fase inicial con la fase final del experimento se aprecia diferencias significativas (p ≤ 0,05) en las mismas zonas, con un predominio de la zona proximal y distal. CONCLUSIONES: en las tres zonas del lóbulo el comportamiento de la densidad superficial de núcleos de hepatocitos ocurre de manera similar, con un sentido al incremento al final del experimento, lo que se corroboró con la aplicación de la prueba de la t de Student. Los incrementos más significativos ocurrieron en la zona proximal y distal, aunque en todas las zonas existen diferencias significativas entre los valores obtenidos en la primera mitad de tiempo del experimento respecto a la parte final.
BACKGROUND: hepatic regeneration is the basic answer of the liver to various internal and external stimuli. It is multifactorial and induced and originates sequential changes in the gene expression, the structure and organization of hepatic cells. OBJECTIVE: to analyze the behaviour of the superficial density of the nuclei of the hepatocytes according to the position they have in the areas of the middle hepatic lobes of mice subjected to regenerative cytosol. METHOD: the superficial density of the nuclei of the hepatocytes was studied with the use of morphometry in 14 middle hepatic lobes of mice subjected to a dose of regenerative liver cytosol at the beginning of the experiment. The samples were taken every 12 hours during eight days. The cut was divided into proximal, medial and distal areas to the hilum, where the determinations were made. Results: the average values and their standard deviations in the areas were: proximal 701,23 ± 135,83 (μm2.103), medial 707,48 ± 46,87 (μm2.103) y distal 682,48 ± 125,21 (μm2.103). When comparing the results through the analysis of variance, it could be seen that there are not differences among the averages in the three areas. When comparing the initial stage with the final stage of the experiment major differences (p ≤ 0,05) could be seen in the same areas, predominating the proximal and distal area. CONCLUSIONS: in the three areas of the lobe the behaviour of the superficial density of the nuclei of the hepatocytes is similar, with a tendency to increase at the end of the experiment. This could be corroborated with the application of the Student’s t- test. In spite of the fact that there were major differences between the values obtained in the first part and the final part of the experiment in all the areas, the most significant increases occurred in the proximal and distal areas.
Subject(s)
Animals , Rats , Hepatocytes/cytology , Liver Regeneration/physiology , Animals, Laboratory , Cell CountABSTRACT
Estudo prospectivo, tipo coorte que visou identificar a influência da sobrecarga de trabalho da Enfermagem na ocorrência de incidentes sem lesão e eventos adversos em 399 pacientes internados em Unidades de Terapia Intensiva (UTI). Para coleta de dados, foi aplicado um questionário estruturado e feita análise de prontuários. Nessas admissões, aproximadamente 78% dos incidentes sem lesão e eventos adversos em pacientes foram relacionados à esfera da Enfermagem. Essas ocorrências foram atribuídas à sobrecarga de trabalho, aumentaram o número de dias de internação e o risco de óbito dos pacientes estudados. É fundamental que os gerentes de enfermagem atuem no processo de gestão de pessoas no âmbito hospitalar, evitando a sobrecarga de trabalho e proporcionando, consequentemente, aumento da segurança do paciente.
This prospective cohort study aimed to identify the influence of nursing work overload on the occurrence of incidents without injury and adverse events in 399 patients hospitalized in Intensive Care Units (ICU). For data collection, a structured questionnaire was administered and an analysis of medical records was performed. In these admissions, approximately 78% of incidents without injury and adverse events in patients were related to the sphere of Nursing. These occurrences were attributed to overwork, increased the number of days of hospitalization and the risk of death of patients. It is essential that nursing managers work on the staff hospital management avoiding work overload to contribute for patient safety.
Estudio de cohorte prospectivo que tuvo como objetivo identificar la influencia de la carga de trabajo de enfermería en la ocurrencia de incidentes sin lesión y los eventos adversos en 399 pacientes ingresados en Unidades de Cuidados Intensivos (UCI). Para la recolección de datos, se administró un cuestionario estructurado y realizó el análisis de los registros en prontuarios. En estos ingresos, aproximadamente el 78% de los incidentes sin lesión y los eventos adversos en los pacientes estaban relacionados con el ámbito de la Enfermería. Estos hechos fueron atribuidos al exceso de trabajo, aumentaran el número de días de hospitalización y el riesgo de muerte de los pacientes. Es esencial que los responsables de enfermería que actúan en el proceso de gestión de las personas en los hospitales, eviten el exceso de trabajo y por lo tanto proporcionar una mayor seguridad de los pacientes.
Subject(s)
Animals , Rats , Drugs, Chinese Herbal/pharmacology , Hepatocytes/drug effects , Apoptosis/drug effects , Caspases/metabolism , Cell Division/drug effects , Cell Line , Enzyme Activation/drug effects , Hepatocytes/cytology , Liver Cirrhosis/drug therapy , Phytotherapy , Salvia miltiorrhizaABSTRACT
The technique of stem cells or hepatocytes transplantation has recently improved in order to bridge the time before whole-organ liver transplantation. In the present study, unfractionated bone marrow stem cells (BMSCs) were harvested from the tibial and femoral marrow compartments of male mice, which were cultured in Dulbecco's modified Eagle's medium (DMEM) with and without hepatocyte growth factor (HGF), and then transplanted into Schistosoma mansoni-infected female mice on their 8th week post-infection. Mice were sacrificed monthly until the third month of bone marrow transplantation, serum was collected, and albumin concentration, ALT, AST, and alkaline phosphatase (ALP) activities were assayed. On the other hand, immunohistopathological and immunohistochemical changes of granuloma size and number, collagen content, and cells expressing OV-6 were detected for identification of liver fibrosis. BMSCs were shown to differentiate into hepatocyte-like cells. Serum ALT, AST, and ALP were markedly reduced in the group of mice treated with BMSCs than in the untreated control group. Also, granuloma showed a marked decrease in size and number as compared to the BMSCs untreated group. Collagen content showed marked decrease after the third month of treatment with BMSCs. On the other hand, the expression of OV-6 increased detecting the presence of newly formed hepatocytes after BMSCs treatment. BMSCs with or without HGF infusion significantly enhanced hepatic regeneration in S. mansoni-induced fibrotic liver model and have pathologic and immunohistopathologic therapeutic effects. Also, this new therapeutic trend could generate new hepatocytes to improve the overall liver functions.
Subject(s)
Animals , Female , Male , Mice , Alanine Transaminase/blood , Alkaline Phosphatase/blood , Antigens, Differentiation/biosynthesis , Aspartate Aminotransferases/blood , Bone Marrow Cells/cytology , Bone Marrow Transplantation , Cell Differentiation , Cell- and Tissue-Based Therapy , Cells, Cultured , Collagen/metabolism , Granuloma/parasitology , Hepatocyte Growth Factor/pharmacology , Hepatocytes/cytology , Liver/parasitology , Liver Cirrhosis/parasitology , Mice, Inbred BALB C , Schistosoma mansoni/pathogenicity , Schistosomiasis mansoni/mortality , Stem Cell Transplantation , Stem Cells/cytologyABSTRACT
Hepatic oval cells (HOCs) are recognized as facultative liver progenitor cells that play a role in liver regeneration after acute liver injury. Here, we investigated the in vitro proliferation and differentiation characteristics of HOCs in order to explore their potential capacity for intrahepatic transplantation. Clusters or scattered HOCs were detected in the portal area and interlobular bile duct in the liver of rats subjected to the modified 2-acetylaminofluorene and partial hepatectomy method. Isolated HOCs were positive for c-kit and CD90 staining (99.8% and 88.8%, respectively), and negative for CD34 staining (3.6%) as shown by immunostaining and flow cytometric analysis. In addition, HOCs could be differentiated into hepatocytes and bile duct epithelial cells after leukemia inhibitory factor deprivation. A two-cuff technique was used for orthotopic liver transplantation, and HOCs were subsequently transplanted into recipients. Biochemical indicators of liver function were assessed 4 weeks after transplantation. HOC transplantation significantly prolonged the median survival time and improved the liver function of rats receiving HOCs compared to controls (P=0.003, Student t-test). Administration of HOCs to rats also receiving liver transplantation significantly reduced acute allograft rejection compared to control liver transplant rats 3 weeks following transplantation (rejection activity index score: control=6.3±0.9; HOC=3.5±1.5; P=0.005). These results indicate that HOCs may be useful in therapeutic liver regeneration after orthotopic liver transplantation.
Subject(s)
Animals , Female , Male , Rats , Cell Proliferation , Cell Differentiation/physiology , Cell Transplantation/methods , Hepatocytes/cytology , Liver Transplantation/methods , Flow Cytometry , Graft Rejection/diagnosis , Hepatectomy , Immunohistochemistry , Liver/anatomy & histology , Liver/surgery , Primary Cell Culture , Rats, Inbred Lew , Real-Time Polymerase Chain Reaction/methods , Survival RateABSTRACT
Embora sejam as forrageiras mais importantes para a pecuária de corte (bovinocultura de corte) no Brasil, em certas épocas ou condições, Brachiaria spp. podem ser tóxicas e causar surtos de fotossensibilização hepatógena que determinam significativas perdas econômicas. Animais que se alimentam em pastos de Brachiaria spp. comumente apresentam macrófagos espumosos isolados ou agrupados, além de cristais no interior de ductos biliares. Saponinas esteroidais têm sido identificadas nestes cristais e são responsabilizadas por lesar o fígado levando ao acúmulo de filoeritrina. Por vezes, imagens negativas desses cristais podem estar presentes no citoplasma de macrófagos espumosos. A patogênese de formação e o tipo de material armazenado nas células espumosas ainda são desconhecidos. A técnica de lectino-histoquímica visa auxiliar na detecção desses macrófagos e, consequentemente, no diagnóstico, além de identificar quais os resíduos de açúcares específicos que estão presentes no citoplasma das células espumosas. Assim, este trabalho teve por objetivo identificar quais lectinas são mais indicadas na detecção de saponinas esteroidais no fígado e rim de ovinos com fotossensibilização causada por Brachiaria decumbens. Fragmentos de fígado e rim de quinze ovinos, de ambos os sexos e idade variável, oriundos de Conceição do Mato Dentro, Minas Gerais, e um ovino mantido em pastagem livre de Brachiaria spp. foram avaliados pela técnica de lectino-histoquímica. Quatorze lectinas foram utilizadas (Con-A, DBA, SBA, PNA, SJA, RCA-I, UEA-I, WGA, SWGA, GSL, PSA, PHA-L, PHA-E e LCA). Verificou-se que, no fígado de ovinos com fotossensibilização provocada pela ingestão de Brachiaria decumbens, a lectina PNA apresentou especificidade e acentuada reatividade aos macrófagos espumosos, bem como especificidade e leve reatividade aos hepatócitos; a lectina WGA teve especificidade e moderada reatividade aos macrófagos espumosos do fígado e especificidade e leve reatividade aos...
Inspite that Brachiaria spp. are most important forage grasses for beef cattle raising in Brazil, they can under certain conditions be toxic, cause outbreaks of hepatogenic photosensibilization and determine severe economic losses. Animals grazing toxic Brachiaria spp. commonly show in their liver foamy macrophages, isolated or grouped together, and crystals inside the biliary ducts. Steroidal saponins have been identified in these crystals and are liable for damage the liverleading to accumulation of phylloerytrin. Occasionally negative images of the crystals may be seen in the cytoplasm of foamy macrophages. The pathogenesis of formation and the type of material stored in the foamy cells is still unknown. The lectin histochemistry aims to assist in the detection of foamy macrophages and consequently in the diagnosis of the poisoning. Lectin histochemistry detects what kind of specific sugar residues exists in the cytoplasm of the foamy cells. The purpose of the present study was to identify the most suitable lectins the detection of steroidal saponins in the liver and kidney of sheep with photosensitization caused by Brachiara decumbens. Fragments of liver and kidney of fifteen sheep of both sexes and variable age, from Conceição do Mato Dentro, Minas Gerais, southeastern Brazil, and one sheep kept free grazing Brachiaria spp. were evaluated by lectin histochemistry. Fourteen lectins were used (Con-A, DBA, SBA, PNA, SJA, RCA-I, UEA-I, WGA, SWGA, GSL, PSA, PHA-L, PHA-E and LCA). It was found that the PNA lectin detected in the liver of sheep with photosensitization, caused by ingestion of Brachiaria decumbens, showed specificity and great binding reactivity for foamy macrophages, and also specificity and slight binding reactivity for hepatocytes. The WGA lectin showed specificity and moderate binding reactivity for foamy macrophages of the liver, and also specificity and slight binding reactivity for hepatocytes. The SBA, GSL and LCA lectins showed...
Subject(s)
Animals , Brachiaria , Liver/anatomy & histology , Hepatocytes/cytology , Kidney/anatomy & histology , Sheep/metabolism , Photosensitivity Disorders/veterinaryABSTRACT
Hepatic progenitor cells (HPCs) are a potential cell source for liver cell transplantation but do not function like mature liver cells. We sought an effective and reliable method to induce HPC maturation. An immortalized HP14.5 albumin promoter-driven Gaussian luciferase (ALB-GLuc) cell line was established from HPCs isolated from fetal mouse liver of post coitus day 14.5 mice to investigate the effect of induction factors on ALB promoter. HP14.5 parental cells were cultured in DMEM with different combinations of 2% horse serum (HS), 0.1 µM dexamethasone (DEX), 10 ng/mL hepatic growth factor (HGF), and/or 20 ng/mL fibroblast growth factor 4 (FGF4). Trypan blue and crystal violet staining were used to assess cell proliferation with different induction conditions. Expression of hepatic markers was measured by semi-quantitative RT-PCR, Western blot, and immunofluorescence. Glycogen storage and metabolism were detected by periodic acid-Schiff and indocyanine green (ICG) staining. GLuc activity indicated ALB expression. The combination of 2% HS+0.1 µM Dex+10 ng/mL HGF+20 ng/mL FGF4 induced the highest ALB-GLuc activity. Cell proliferation decreased in 2% HS but increased by adding FGF4. Upon induction, and consistent with hepatocyte development, DLK, AFP, and CK19 expression decreased, while ALB, CK18, and UGT1A expression increased. The maturity markers tyrosine aminotransferase and apolipoprotein B were detected at days 3 and 6 post-induction, respectively. ICG uptake and glycogen synthesis were detectable at day 6 and increased over time. Therefore, we demonstrated that HPCs were induced to differentiate into functional mature hepatocytes in vitro, suggesting that factor-treated HPCs may be further explored as a means of liver cell transplantation.
Subject(s)
Animals , Mice , Cell Differentiation/drug effects , Embryo, Mammalian/drug effects , Hepatocytes/cytology , Liver/cytology , Stem Cells/drug effects , Antigens, Differentiation/analysis , Apolipoproteins B/isolation & purification , Cell Proliferation , Dexamethasone/administration & dosage , Fibroblast Growth Factors/administration & dosage , Gentian Violet , Glycogen/metabolism , Hepatocyte Growth Factor/administration & dosage , Indocyanine Green/pharmacokinetics , Primary Cell Culture/methods , Reverse Transcriptase Polymerase Chain Reaction , Stem Cells/cytology , Trypan Blue , Tyrosine Transaminase/isolation & purificationABSTRACT
Vitamin D3 up-regulated protein 1 (VDUP1) is a potent growth suppressor that inhibits tumor cell proliferation and cell cycle progression when overexpressed. In a previous study, we showed that VDUP1 knockout (KO) mice exhibited accelerated liver regeneration because such animals could effectively control the expression of cell cycle regulators that drive the G1-to-S phase progression. In the present study, we further investigated the role played by VDUP1 in initial priming of liver regeneration. To accomplish this, VDUP1 KO and wild-type (WT) mice were subjected to 70% partial hepatectomy (PH) and sacrificed at different times after surgery. The hepatic levels of TNF-alpha and IL-6 increased after PH, but there were no significant differences between VDUP1 KO and WT mice. Nuclear factor-kappaB (NF-kappaB), c-Jun-N-terminal kinase (JNK), and signal transducer and activator of transcription 3 (STAT-3) were activated much earlier and to a greater extent in VDUP1 KO mice after PH. A single injection of TNF-alpha or IL-6 caused rapid activation of JNK and STAT-3 expression in both mice, but the responses were stronger and more sustained in VDUP1 KO mice. In conclusion, our findings provide evidence that VDUP1 plays a role in initiation of liver regeneration.
Subject(s)
Animals , Male , Blotting, Western , Carrier Proteins/genetics , Cell Proliferation , Gene Expression Regulation , Hepatectomy , Hepatocytes/cytology , JNK Mitogen-Activated Protein Kinases/genetics , Liver/physiology , Mice, Knockout , NF-kappa B/genetics , Polymerase Chain Reaction , Regeneration , STAT3 Transcription Factor/genetics , Thioredoxins/geneticsABSTRACT
Este trabalho teve como objetivo descrever macro e microscopicamente o fígado do Tambaqui Colossoma macropomum, Teleósteo de água doce da Família Characidae, de grande interesse econômico da bacia Amazônica. Foram utilizados seis (6) exemplares jovens com idade entre seis meses e um ano, oriundos da Chácara Esteio, Alta Floresta, MT, que desenvolve principalmente a prática da piscicultura. O órgão foi fotodocumentado in situ e descrito macroscopicamente, em seguida procedeu-se a retirada de fragmentos deste, que foram processados pelas técnicas histológicas rotineiras para inclusão em parafina e coloração de HE. O fígado localizou-se ventral à bexiga natatória e craniodorsalmente ao estômago, apresentou coloração amarronzada a vermelho, constituído por três lobos hepáticos, o lobo lateral direito, o lobo lateral esquerdo e o lobo ventral. Microscopicamente, o parênquima era constituído por hepatócitos com formato que variou do arredondado irregular hexagonal ao redondo com núcleo grande e central, arranjados em cordões lineares limitados por sinusóides que irradiam para veias centrais, e com ausência de lóbulos hepáticos. As veias centrais estavam distribuídas pelo parênquima, enquanto que o espaço porta, na maioria das vezes, era constituído apenas por uma veia hepática e o ducto biliar, em outros locais foi observado, uma artéria e um ducto. Não foi observada a formação de tríades portais. Foram frequentemente observados melano macrófagos centrais dispersos pelo parênquima. O estudo morfofuncional do Aparelho Digestório de peixes da bacia Amazônica, se faz pertinente com vistas ao conhecimento do aproveitamento de ganho de peso e produção em alta escala para consumo humano e preservação da espécie, além da importância de estarem sendo utilizados como bioindicadores atualmente.
This research aimed to describe the macroscopic and microscopic liver of tambaqui, Colossoma macropomum, Teleost freshwater Family Characidae, of great economic interest for the Amazon basin. We used six juveniles aged between six month and one year, from the small holding Esteio, Alta Floresta/MT, that develops mainly fish farming. The body was photographed in situ, described macroscopically, and fragments were removed and processed by routine histological techniques through paraffin embedding and HE staining. The liver, located ventrally to the swim bladder and craniodorsally to the stomach, is brownish red and consisted of three lobes, the right lateral, the left lateral and the ventral lobe. Microscopically, the parenchyma consists of hepatocytes varying from irregular rounded hexagonal to round forms with a large and central nucleus, and arranged in linear strings limited by sinusoids and radiating to central veins, but with absence of liver lobules. The central veins are distributed throughout the parenchyma, while the portal space consists in most cases only of a hepatic vein and bile duct; elsewhere exist artery and duct. Formation of portal triads was not founde. Melano macrophages were frequently seen dispersed throughout the central parenchyma. The morphofunctional study of the digestive system of fishes of the Amazon basin is important to obtain knowledge about their weight gain, large scale production for human consumption and preservation of the species, and has also its importance for being used as bioindicators today.
Subject(s)
Animals , Characidae/physiology , Liver/anatomy & histology , Hepatocytes/cytology , Fishes/physiology , Portal Vein/physiologyABSTRACT
The flower of Butea monosperma (Lam.) (Fabaceae) has been used in traditional Indian medicine in the treatment of many ailments including liver disorders. To understand the pharmacological basis of its beneficial effects, the extracts of dried flowers in water, methanol, butanol, ethyl acetate and acetone were evaluated for free radical scavenging and pro-apoptotic activities in cell cultures (human hepatoma Huh-7 cell line and immortalized AML-12 mouse hepatocytes). Butrin and butein -the active constituents of flower extracts- were used as reference molecules. The levels of cell injury markers like lactate dehydrogenase, glutathione and lipid peroxidation and primary antioxidant enzymes glutathione S-transferase and catalase were also measured. The aqueous and butanolic extracts exhibited better 2,2-diphenyl-1-picrylhydrazyl scavenging and cytotoxic activities in hepatoma cells than in immortalized hepatocytes. Interestingly, butein inhibited 2,2-diphenyl-1-picrylhydrazyl radical better than butrin. The aqueous and butanolic extracts were further investigated for hepatoprotection against carbon tertrachloride-induced biochemical changes and cell death. Both extracts, just as butrin and butein, significantly reversed the cellular glutathione levels and lipid peroxidation, and glutathione-S-transferase activity. Lactate dehydrogenase leakage and cell death were also prevented. However, only butein revived the catalase activity. Thus, the butein content of Butea monosperma flower extracts is important for free radical scavenging activity, apoptotic cell death and protection against oxidative injury in hepatic cells.
Subject(s)
Animals , Humans , Mice , Antioxidants/pharmacology , Apoptosis/drug effects , Butea/chemistry , Chalcones/pharmacology , Flowers/chemistry , Free Radical Scavengers/pharmacology , Free Radicals/metabolism , Plant Extracts/pharmacology , Antioxidants/isolation & purification , Cells, Cultured , Carcinoma, Hepatocellular/drug therapy , Carcinoma, Hepatocellular/metabolism , Carcinoma, Hepatocellular/pathology , Chalcones/isolation & purification , Flavonoids/isolation & purification , Flavonoids/pharmacology , Free Radical Scavengers/isolation & purification , Glutathione Transferase/metabolism , Glutathione/metabolism , Hepatocytes/cytology , Hepatocytes/drug effects , Hepatocytes/metabolism , Lipid Peroxidation/drug effects , Liver Neoplasms/drug therapy , Liver Neoplasms/metabolism , Liver Neoplasms/pathology , Oxidation-ReductionABSTRACT
Generally, extra-cellular-signal-regulated kinase 5 (ERK5) signaling pathway regulates many physiological activities, such as cell proliferation and cell differentiation. However, little is known about how ERK5 signaling pathway composed of 15 paths participates in regulating hepatocyte proliferation during liver regeneration (LR). In this study, to explore the influence ERK5 signaling pathway upon hepatocytes at gene transcription level, rat genome 230 2.0 array was used to detect expression changes of 75 related genes in isolated hepatocytes from rat regenerating liver. Bioinformatics and systems biology methods were applied to analyze the precise role of ERK5 signaling pathway in regulating hepatocyte proliferation during LR. Results showed that 62 genes were contained in the array and 22 genes were significantly changed. It was found that 6 paths were related to hepatocyte proliferation during rat LR. Among them, paths 3, 6 and 13 of ERK5 signaling pathway modulated cell cycle progression by decreasing the negative influence on ERK5 and paths 3, 4, 8 and 9 by reinforcing the positive influence on ERK5. In summary, the study shows that 22 genes and 6 paths of ERK5 signaling pathway participate in regulating proliferation of hepatocytes in rat LR.
Subject(s)
Animals , Cell Growth Processes/genetics , Cell Growth Processes/physiology , Gene Expression Profiling/methods , Hepatectomy , Hepatocytes/cytology , Hepatocytes/enzymology , Hepatocytes/physiology , Liver Regeneration/genetics , Liver Regeneration/physiology , MAP Kinase Signaling System/genetics , MAP Kinase Signaling System/physiology , Mitogen-Activated Protein Kinase 7/genetics , Mitogen-Activated Protein Kinase 7/metabolism , Oligonucleotide Array Sequence Analysis/methods , Random Allocation , Rats , Rats, Sprague-DawleyABSTRACT
The role of ion channels and particularly cationic channels in the pathogenesis of various diseases are being considered carefully. The diabetes mellitus is a common disease which is initiated by ion channel disturbances. This study was done to determine the characteristics of hepatocyte rough endoplasmic reticulum single cationic channel in Streptozocin-induced diabetic rats. This experimental study was done on 10 male adult Wistar rats and animals were randomly allocaied into diabetic and control groups. Diabetes induced by STZ [65 mg/kg/bw] intraperitounally. Rough endoplasmic reticulum vesicles were extracted following rat liver excision, homogenization and ultracentrifuging. The bilayer membrane formation was prepared by painting phosphatidylcholine on 250 micro M aperture in between Cis and Trans sides. The RER vesicles incorporation was performed through gentle and delicate touch of membrane using a dentistry needle. The Pclamp9 software was used for ion channel activity characteristic analysis. The cationic channel current amplitude did not change significantly in voltages more than+3o mV but their open probability [Po] decreased in diabetic group [P<0.05]. More severe changes in channel activity were seen in potentials less than the reverse potential. In addition to significant increase of channel Po [P<0.05], also, the channel unitary currents were significantly decreased [P<0.05]. The mean current amplitude and channel open probability in voltage+40 mV were 17 +/- 2.14 pA and 0.68 +/- 0.01 in control group respectively, whereas, the values of these parameters reached to 18.5 +/- 2.5 and 0.26 +/- 0.03, respectively. In voltage-10 mV, the values of mean current amplitude and Po were-22.3 +/- 2.14 pA and<0.1 in control group, respectively but the values changed to-13.1 +/- 0.08 and 0.62 +/- 0.03 in diabetic group. It seems that RER cationic channel is involved in metabolic changes which cause by diabetes mellitus and this disease can cause probably a channel gating kinetic and behavior change by inducing metabolic stresses