RESUMEN
Mesenchymal stem cell (MSC)-mediated immunomodulation has been harnessed for the treatment of human diseases, but its underlying mechanism has not been fully understood. Dead cells, including apoptotic cells have immunomodulatory properties. It has been repeatedly reported that the proportion of nonviable MSCs in a MSC therapeutic preparation varied from 5~50% in the ongoing clinical trials. It is conceivable that the nonviable cells in a MSC therapeutic preparation may play a role in the therapeutic effects of MSCs. We found that the MSC therapeutic preparation in the present study had about 5% dead MSCs (DMSCs), characterized by apoptotic cells. Namely, 1 × 106 MSCs in the preparation contained about 5 × 104 DMSCs. We found that the treatment with even 5 × 104 DMSCs alone had the equal therapeutic effects as with 1 × 106 MSCs. This protective effect of the dead MSCs alone was confirmed in four mouse models, including concanavalin A (ConA)- and carbon tetrachloride (CCl4)-induced acute liver injury, LPS-induced lung injury and spinal cord injury. We also found that the infused MSCs died by apoptosis in vivo. Furthermore, the therapeutic effect was attributed to the elevated level of phosphatidylserine (PS) upon the injection of MSCs or DMSCs. The direct administration of PS liposomes (PSLs) mimic apoptotic cell fragments also exerted the protective effects as MSCs and DMSCs. The Mer tyrosine kinase (MerTK) deficiency or the knockout of chemokine receptor C-C motif chemokine receptor 2 (CCR2) reversed these protective effects of MSCs or DMSCs. These results revealed that DMSCs alone in the therapeutic stem cell preparation or the apoptotic cells induced in vivo may exert the same immunomodulatory property as the "living MSCs preparation" through releasing PS, which was further recognized by MerTK and participated in modulating immune cells.
Asunto(s)
Apoptosis , Intoxicación por Tetracloruro de Carbono , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/metabolismo , Fosfatidilserinas/metabolismo , Animales , Intoxicación por Tetracloruro de Carbono/metabolismo , Intoxicación por Tetracloruro de Carbono/terapia , Femenino , Ratones , Ratones NoqueadosRESUMEN
Bidens pilosa L. is a medicinal plant popularly used for treatment of liver diseases. In this study, the dry extract of aerial parts of Bidens pilosa and Silymarin, a phytocomplex obtained from the Silybum marianum fruits and marketed as hepatoprotective, were tested in dogs experimentally acutely intoxicated with carbon tetrachloride. The liver activity was evaluated by hematological and biochemical profiles, and histological and ultrasound analyzes. It was observed that the lowest serum activities of ALT and serum concentrations of total bilirubin occurred in the groups treated with the dry extract of Bidens pilosa, while only decreased serum concentrations of total bilirubin occurred in the group treated with Silymarin. Best liver recovery was also observed for the dry extract of B. pilosa at a 400mg/Kg dose by ultrasonography. This study showed that the dry extract of Bidens pilosa acted more efficiently in the treatment of acute toxic hepatitis induced in dogs than Silymarin.(AU)
Bidens pilosa L. é uma planta medicinal utilizada popularmente para tratamento de doenças hepáticas. Neste trabalho, o extrato seco das partes aéreas da Bidens pilosa e a silimarina, um fitocomplexo obtido dos frutos da Silybum marianum e comercializado como hepatoprotetor, foram testados em cães intoxicados experimentalmente de forma aguda com tetracloreto de carbono. A atividade hepática foi avaliada por meio dos perfis hematológico e bioquímico, análises histológica e ultrassonográfica. Observou-se que, nos grupos tratados com o extrato seco da Bidens pilosa, ocorreram as menores atividades séricas da ALT e de concentrações séricas de bilirrubina total, enquanto no grupo tratado com silimarina, ocorreu apenas diminuição de concentrações séricas de bilirrubina total. Melhor recuperação hepática também foi verificada para o extrato seco de B. pilosa na dose de 400mg/kg por ultrassonografia. Este estudo evidenciou que o extrato seco da Bidens pilosa atuou de forma mais eficiente no tratamento da hepatite aguda tóxica induzida em cães do que a silimarina.(AU)
Asunto(s)
Animales , Perros , Intoxicación por Tetracloruro de Carbono/terapia , Intoxicación por Tetracloruro de Carbono/veterinaria , Bidens/química , Hepatitis Animal/terapia , Plantas Medicinales , Silimarina/uso terapéuticoRESUMEN
Direct conversion from fibroblasts to generate hepatocyte like-cells (iHeps) bypassing the pluripotent state has been described in previous reports as an attractive method acquiring hepatocytes for cell-based therapy. The limited proliferation of iHeps, however, has hampered it uses in cell-based therapy. Since hepatic stem cells (HepSCs) possess self-renewal and bipotency with the capacity to differentiate into both hepatocytes and cholangiocytes, they have therapeutic potential for treating liver disease. Here, we investigated the therapeutic effects of induced HepSCs (iHepSCs) on a carbon tetrachloride (CCl4)-induced liver fibrosis model. We demonstrate that Oct4 and Hnf4a are sufficient to convert fibroblasts into expandable iHepSCs. Hepatocyte-like cells derived from iHepSCs (iHepSC-HEPs) exhibit the typical morphology of hepatocytes and hepatic functions, including glycogen storage, low-density lipoprotein (LDL) uptake, Indocyanine green (ICG) detoxification, drug metabolism, urea production, and albumin secretion. iHepSCs-derived cholangiocyte-like cells (iHepSC-CLCs) expressed cholangiocyte-specific markers and formed cysts and tubule-like structures with apical-basal polarity and secretory function in three-dimensional culture condition. Furthermore, iHepSCs showed anti-inflammatory and anti-fibrotic effects in CCl4-induced liver fibrosis. This study demonstrates that Oct4 and Hnf4α-induced HepSCs show typical hepatic and biliary functionality in vitro. It also presents the therapeutic effect of iHepSCs in liver fibrosis. Therefore, directly converting iHepSCs from somatic cells may facilitate the development of patient-specific cell-based therapy for chronic liver damage.
Asunto(s)
Intoxicación por Tetracloruro de Carbono , Factor Nuclear 4 del Hepatocito , Células Madre Pluripotentes Inducidas , Cirrosis Hepática , Hígado , Lesión Pulmonar , Factor 3 de Transcripción de Unión a Octámeros , Trasplante de Células Madre , Animales , Intoxicación por Tetracloruro de Carbono/genética , Intoxicación por Tetracloruro de Carbono/metabolismo , Intoxicación por Tetracloruro de Carbono/terapia , Factor Nuclear 4 del Hepatocito/genética , Factor Nuclear 4 del Hepatocito/metabolismo , Células Madre Pluripotentes Inducidas/metabolismo , Células Madre Pluripotentes Inducidas/patología , Células Madre Pluripotentes Inducidas/trasplante , Hígado/metabolismo , Hígado/patología , Cirrosis Hepática/inducido químicamente , Cirrosis Hepática/genética , Cirrosis Hepática/metabolismo , Cirrosis Hepática/terapia , Lesión Pulmonar/inducido químicamente , Lesión Pulmonar/genética , Lesión Pulmonar/metabolismo , Masculino , Ratones , Factor 3 de Transcripción de Unión a Octámeros/genética , Factor 3 de Transcripción de Unión a Octámeros/metabolismoRESUMEN
BACKGROUND: Mesenchymal stem cells (MSCs) are increasingly being applied as a therapy for liver fibrosis. Exosomes possess similar functions to their parent cells; however, they are safe and effective cell-free reagents with controllable and predictable outcomes. In this study, we investigated the therapeutic potential and underlying molecular mechanism for human bone mesenchymal stem cells-derived exosomes (hBM-MSCs-Ex) in the treatment of liver fibrosis. METHODS: We established an 8-week CCl4-induced rat liver fibrosis model, after which, we administered hBM-MSCs-Ex in vivo for 4 weeks. The resulting histopathology, liver function, and inflammatory cytokines were analyzed. In addition, we investigated the anti-fibrotic mechanism of hBM-MSCs-Ex in hepatic stellate cells (HSCs) and liver fibrosis tissue, by western blotting for the expression of Wnt/ß-catenin signaling pathway-related genes. RESULTS: In vivo administration of hBM-MSCs-Ex effectively alleviated liver fibrosis, including a reduction in collagen accumulation, enhanced liver functionality, inhibition of inflammation, and increased hepatocyte regeneration. Moreover, based on measurement of the collagen area, Ishak fibrosis score, MDA levels, IL-1, and IL-6, the therapeutic effect of hBM-MSCs-Ex against liver fibrosis was significantly greater than that of hBM-MSCs. In addition, we found that hBM-MSCs-Ex inhibited the expression of Wnt/ß-catenin pathway components (PPARγ, Wnt3a, Wnt10b, ß-catenin, WISP1, Cyclin D1), α-SMA, and Collagen I, in both HSCs and liver fibrosis tissue. CONCLUSIONS: These results suggest that hBM-MSCs-Ex treatment could ameliorate CCl4-induced liver fibrosis via inhibition of HSC activation through the Wnt/ß-catenin pathway.
Asunto(s)
Células de la Médula Ósea/metabolismo , Intoxicación por Tetracloruro de Carbono , Exosomas , Cirrosis Hepática , Células Madre Mesenquimatosas/metabolismo , Vía de Señalización Wnt , Animales , Células de la Médula Ósea/patología , Intoxicación por Tetracloruro de Carbono/metabolismo , Intoxicación por Tetracloruro de Carbono/patología , Intoxicación por Tetracloruro de Carbono/terapia , Exosomas/metabolismo , Exosomas/patología , Exosomas/trasplante , Femenino , Xenoinjertos , Humanos , Cirrosis Hepática/metabolismo , Cirrosis Hepática/patología , Cirrosis Hepática/terapia , Células Madre Mesenquimatosas/patología , Ratas , Ratas Sprague-Dawley , beta CateninaRESUMEN
Hepatic stellate cells (HSCs) are the primary cell type responsible for liver fibrogenesis. Transforming growth factor beta 1 (TGF-ß1) and platelet-derived growth factor (PDGF) are key profibrotic cytokines that regulate HSC activation and proliferation with functional convergence. Dual RNA interference against their receptors may achieve therapeutic effects. A novel RNAi strategy based on HSC-specific GFAP promoter-driven and lentiviral-expressed artificial microRNAs (amiRNAs) was devised that consists of an microRNA-30a backbone and effective shRNAs against mouse Pdgfrß and Tgfbr2. Then, its antifibrotic efficacy was tested in primary and cultured HSCs and in mice affected with carbon tetrachloride-induced hepatic fibrosis. The study shows that amiRNA-mediated Pdgfrß and Tgfbr2 co-silencing inhibits HSC activation and proliferation. After recombinant lentiviral particles were delivered into the liver via tail-vein injection, therapeutic amiRNAs were preferentially expressed in HSCs and efficiently co-knocked down in situ Tgfbr2 and Pdgfrß expression, which correlates with downregulated expression of target or effector genes of their signaling, which include Pai-1, P70S6K, and D-cyclins. amiRNA-based HSC-specific co-silencing of Tgfbr2 and Pdgfrß significantly suppressed hepatic expression of fibrotic markers α-Sma and Col1a1, extracellular matrix regulators Mmps and Timp1, and phenotypically ameliorated liver fibrosis, as indicated by reductions in serum alanine aminotransferase activity, collagen deposition, and α-Sma-positive staining. The findings provide proof of concept for the use of amiRNA-mediated co-silencing of two profibrogenic pathways in liver fibrosis treatment and highlight the therapeutic potential of concatenated amiRNAs for gene therapy.
Asunto(s)
Intoxicación por Tetracloruro de Carbono , Silenciador del Gen , Cirrosis Hepática , MicroARNs , Receptor beta de Factor de Crecimiento Derivado de Plaquetas , Receptor Tipo II de Factor de Crecimiento Transformador beta , Animales , Intoxicación por Tetracloruro de Carbono/genética , Intoxicación por Tetracloruro de Carbono/metabolismo , Intoxicación por Tetracloruro de Carbono/patología , Intoxicación por Tetracloruro de Carbono/terapia , Línea Celular , Células Estrelladas Hepáticas/metabolismo , Células Estrelladas Hepáticas/patología , Cirrosis Hepática/genética , Cirrosis Hepática/metabolismo , Cirrosis Hepática/patología , Cirrosis Hepática/terapia , Ratones , MicroARNs/biosíntesis , MicroARNs/genética , Ratas , Ratas Sprague-Dawley , Receptor beta de Factor de Crecimiento Derivado de Plaquetas/biosíntesis , Receptor beta de Factor de Crecimiento Derivado de Plaquetas/genética , Receptor Tipo II de Factor de Crecimiento Transformador beta/biosíntesis , Receptor Tipo II de Factor de Crecimiento Transformador beta/genéticaRESUMEN
Advanced liver disease (ALD) is often characterized with overt malnutrition and liver fibrosis. In this study, a dietary supplement (DS) was first developed, including branch chain amino acids, fat soluble vitamins, zinc, medium chain triglycerides, soy lecithin, L-carnitine, and n-3 polyunsaturated fatty acids. Benefits of DS were then tested using an ALD rat model treated with carbon tetrachloride (CCl4) for 6, 8, and 10 weeks, respectively. Our study showed that CCl4-induced drop of serum albumin and ratio of branch chain to aromatic amino acids were significantly prevented at all three time points. DS also mitigated CCl4-induced elevation of classical liver function markers (alanine aminotransferase, aspartate aminotransferase, and bilirubin) at certain time points, depending on specific liver function markers. Moreover, CCl4-induced liver fibrosis was strongly inhibited at all three time points in a transforming growth factor beta (TGF-ß) independent manner. These findings indicated multi-faceted benefits of DS in this animal model, suggesting that it could be a useful adjunctive treatment of ALD in clinic.
Asunto(s)
Intoxicación por Tetracloruro de Carbono/terapia , Enfermedad Hepática Inducida por Sustancias y Drogas/terapia , Suplementos Dietéticos , Cirrosis Hepática/terapia , Alanina Transaminasa/sangre , Aminoácidos Aromáticos/sangre , Aminoácidos de Cadena Ramificada/administración & dosificación , Aminoácidos de Cadena Ramificada/sangre , Animales , Aspartato Aminotransferasas/sangre , Bilirrubina/sangre , Intoxicación por Tetracloruro de Carbono/sangre , Carnitina/administración & dosificación , Enfermedad Hepática Inducida por Sustancias y Drogas/sangre , Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Ácidos Grasos Omega-3/administración & dosificación , Lecitinas/administración & dosificación , Hígado/metabolismo , Cirrosis Hepática/sangre , Cirrosis Hepática/inducido químicamente , Pruebas de Función Hepática , Ratas , Ratas Sprague-Dawley , Albúmina Sérica/análisis , Glycine max/química , Triglicéridos/administración & dosificación , Vitaminas/administración & dosificación , Zinc/administración & dosificaciónRESUMEN
Adipose-derived stem cells (ADSCs) derived from adipose tissue have the capacity to differentiate into endodermal, mesoderm, and ectodermal cell lineages in vitro, which are an ideal engraft in tissue-engineered repair. In this study, human ADSCs were isolated from subcutaneous fat. The markers of ADSCs, CD13, CD71, CD73, CD90, CD105, CD166, CYP3A4, and ALB were detected by immunofluorescence assays. Human ADSCs were cultured in a specific hepatogenesis differentiation medium containing HGF, bFGF, nicotinamide, ITS, and oncostatin M for hepatogenic differentiation. The hepatocyte markers were analyzed using immunofluorescence and real-time PCR after dramatic changes in morphology. Hepatocytes derived from ADSCs or ADSCs were transplanted into the mice of liver injury for observation cells colonization and therapy in liver tissue. The result demonstrated that human ADSCs were positive for the CD13, CD71, CD73, CD90, CD105, and CD166 but negative for hepatocyte markers, ALB and CYP3A4. After hepatogenic differentiation, the hepatocytes were positive for liver special markers, gene expression level showed a time-lapse increase with induction time. Human ADSCs or ADSCs-derived hepatocyte injected into the vein could improve liver function repair and functionally rescue the CCl4-treated mice with liver injury, but the ADSCs transplantation was better than ADSCs-derived hepatocyte transplantation. In conclusion, our research shows that a population of hepatocyte can be specifically generated from human ADSCs and that cells may allow for participation in tissue-repair.
Asunto(s)
Tejido Adiposo/metabolismo , Intoxicación por Tetracloruro de Carbono/terapia , Hepatocitos , Hígado/metabolismo , Células Madre/metabolismo , Enfermedad Aguda , Animales , Intoxicación por Tetracloruro de Carbono/metabolismo , Hepatocitos/metabolismo , Hepatocitos/trasplante , Xenoinjertos , Humanos , RatonesRESUMEN
To study the effects of keratinocyte growth factor (KGF) modified umbilical cord mesenchymal stem cells (MSC) on chronic liver injury in rats, adenovirus carrying human KGF gene (Ad-KGF) was used. Rat liver injury model was established by subcutaneous injection of CCl4 - olive oil solution; 56 male Wistar rats were randomly divided into normal control, model, MSC, KGF, and KGF/MSC groups. Of all three treatments, KGF/MSC had the most obvious therapeutic effects on liver injury, and cell injections did not cause adverse reaction. The experiment provides new data for clinical research of KGF/MSC and possible methods for liver injury treatment.
Asunto(s)
Adenoviridae , Intoxicación por Tetracloruro de Carbono , Factor 7 de Crecimiento de Fibroblastos , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/metabolismo , Transducción Genética , Animales , Intoxicación por Tetracloruro de Carbono/metabolismo , Intoxicación por Tetracloruro de Carbono/terapia , Enfermedad Crónica , Factor 7 de Crecimiento de Fibroblastos/biosíntesis , Factor 7 de Crecimiento de Fibroblastos/genética , Humanos , Masculino , Ratas , Ratas WistarRESUMEN
The cause of organ failure is enigmatic for many degenerative diseases, including end-stage liver disease. Here, using a CCl4-induced rat model of irreversible and fatal hepatic failure, which also exhibits terminal changes in the extracellular matrix, we demonstrated that chronic injury stably reprograms the critical balance of transcription factors and that diseased and dedifferentiated cells can be returned to normal function by re-expression of critical transcription factors, a process similar to the type of reprogramming that induces somatic cells to become pluripotent or to change their cell lineage. Forced re-expression of the transcription factor HNF4α induced expression of the other hepatocyte-expressed transcription factors; restored functionality in terminally diseased hepatocytes isolated from CCl4-treated rats; and rapidly reversed fatal liver failure in CCl4-treated animals by restoring diseased hepatocytes rather than replacing them with new hepatocytes or stem cells. Together, the results of our study indicate that disruption of the transcription factor network and cellular dedifferentiation likely mediate terminal liver failure and suggest reinstatement of this network has therapeutic potential for correcting organ failure without cell replacement.
Asunto(s)
Redes Reguladoras de Genes , Terapia Genética , Vectores Genéticos/uso terapéutico , Cirrosis Hepática Experimental/terapia , Fallo Hepático/terapia , Factores de Transcripción/fisiología , Animales , Proteína alfa Potenciadora de Unión a CCAAT/biosíntesis , Proteína alfa Potenciadora de Unión a CCAAT/genética , Intoxicación por Tetracloruro de Carbono/genética , Intoxicación por Tetracloruro de Carbono/terapia , Desdiferenciación Celular/genética , Células Cultivadas , Dependovirus/genética , Progresión de la Enfermedad , Regulación hacia Abajo , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Factor Nuclear 1-alfa del Hepatocito/biosíntesis , Factor Nuclear 1-alfa del Hepatocito/genética , Factor Nuclear 3-beta del Hepatocito/biosíntesis , Factor Nuclear 3-beta del Hepatocito/genética , Factor Nuclear 4 del Hepatocito/biosíntesis , Factor Nuclear 4 del Hepatocito/genética , Factor Nuclear 4 del Hepatocito/fisiología , Hepatocitos/metabolismo , Hepatocitos/patología , Cirrosis Hepática Experimental/complicaciones , Cirrosis Hepática Experimental/genética , Cirrosis Hepática Experimental/patología , Fallo Hepático/etiología , Fallo Hepático/genética , Fallo Hepático/patología , Masculino , PPAR alfa/biosíntesis , PPAR alfa/genética , Ratas , Ratas Endogámicas Lew , Proteínas Recombinantes de Fusión/metabolismo , Transcriptoma , Transducción GenéticaRESUMEN
Melatonin's effect on hepatic differentiation of stem cells remains unclear. The aim of this study was to investigate the action of melatonin on hepatic differentiation as well as its related signaling pathways of human dental pulp stem cells (hDPSCs) and to examine the therapeutic effects of a combination of melatonin and hDPSC transplantation on carbon tetrachloride (CCl4 )-induced liver fibrosis in mice. In vitro hepatic differentiation was assessed by periodic acid-Schiff (PAS) staining and mRNA expression for hepatocyte markers. Liver fibrosis model was established by injecting 0.5 mL/kg CCl4 followed by treatment with melatonin (5 mg/kg, twice a week) and hDPSCs. In vivo therapeutic effects were evaluated by histopathology and by means of liver function tests including measurement of alanine transaminase (ALT), aspartate transaminase (AST), and ammonia levels. Melatonin promoted hepatic differentiation based on mRNA expression of differentiation markers and PAS-stained glycogen-laden cells. In addition, melatonin increased bone morphogenic protein (BMP)-2 expression and Smad1/5/8 phosphorylation, which was blocked by the BMP antagonist noggin. Furthermore, melatonin activated p38, extracellular signal-regulated kinase (ERK), and nuclear factor-κB (NF-κB) in hDPSCs. Melatonin-induced hepatic differentiation was attenuated by inhibitors of BMP, p38, ERK, and NF-κB. Compared to treatment of CCl4 -injured mice with either melatonin or hDPSC transplantation alone, the combination of melatonin and hDPSC significantly suppressed liver fibrosis and restored ALT, AST, and ammonia levels. For the first time, this study demonstrates that melatonin promotes hepatic differentiation of hDPSCs by modulating the BMP, p38, ERK, and NF-κB pathway. Combined treatment of grafted hDPSCs and melatonin could be a viable approach for the treatment of liver cirrhosis.
Asunto(s)
Antioxidantes/farmacología , Diferenciación Celular/efectos de los fármacos , Pulpa Dental/metabolismo , Hepatocitos/metabolismo , Cirrosis Hepática/terapia , Melatonina/farmacología , Trasplante de Células Madre , Células Madre/metabolismo , Animales , Antígenos de Diferenciación/biosíntesis , Intoxicación por Tetracloruro de Carbono/metabolismo , Intoxicación por Tetracloruro de Carbono/patología , Intoxicación por Tetracloruro de Carbono/terapia , Pulpa Dental/patología , Regulación de la Expresión Génica/efectos de los fármacos , Hepatocitos/patología , Xenoinjertos , Humanos , Cirrosis Hepática/inducido químicamente , Cirrosis Hepática/metabolismo , Cirrosis Hepática/patología , Masculino , Ratones , Ratones Desnudos , Células Madre/patologíaRESUMEN
Chronic psychological stress has been demonstrated to play an important role in several severe diseases, but whether it affects disease therapy or not remains unclear. Mesenchymal stem cells (MSCs) have been demonstrated to have therapeutic potentials in treating tissue injury based on their multidifferentiation potential toward various cell types. We investigated the effect of chronic restraint stress on therapeutic potential of MSCs on carbon tetrachloride (CCl4)-induced liver injury in mice. CCl4-induced mice were injected with enhanced green fluorescent protein-MSCs, which was followed by chronic restraint stress administration. Corticosterone and RU486, a glucocorticoid receptor (GR) antagonist, were employed in vivo and in vitro, too. In the present study, we illustrated that MSCs could repair liver injury by differentiating into myofibroblasts (MFs) which contribute to fibrosis, whereas stress repressed differentiation of MSCs into MFs displayed by reducing α-smooth muscle actin (α-SMA, a solid marker of MFs) expression. Whereas RU486 could maintain the liver injury reduction and liver fibrosis increases induced by MSCs in stressed mice and block the decrease of α-SMA expression induced by stress. Furthermore, chronic stress inhibited MFs differentiation from MSCs by inhibiting transforming growth factor-ß1 (TGF-ß1)/Smads signaling pathway which is essential for MFs differentiation. Chronic stress reduced autocrine TGF-ß1 of MSCs, but not blunted activation of Smads. All these data suggested that corticosterone triggered by chronic stress impaired liver injury repair by MSCs through inhibiting TGF-ß1 expression which results in reduced MFs differentiation of MSCs.
Asunto(s)
Intoxicación por Tetracloruro de Carbono/terapia , Enfermedad Hepática Inducida por Sustancias y Drogas/terapia , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/metabolismo , Estrés Fisiológico , Factor de Crecimiento Transformador beta1/biosíntesis , Aloinjertos , Animales , Antiinflamatorios/farmacología , Intoxicación por Tetracloruro de Carbono/genética , Intoxicación por Tetracloruro de Carbono/metabolismo , Intoxicación por Tetracloruro de Carbono/patología , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/genética , Enfermedad Hepática Inducida por Sustancias y Drogas/genética , Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Enfermedad Crónica , Corticosterona/metabolismo , Corticosterona/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/genética , Antagonistas de Hormonas/farmacología , Cirrosis Hepática/genética , Cirrosis Hepática/metabolismo , Cirrosis Hepática/patología , Cirrosis Hepática/terapia , Masculino , Células Madre Mesenquimatosas/patología , Ratones , Ratones Transgénicos , Mifepristona/farmacología , Restricción Física , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Proteínas Smad/genética , Proteínas Smad/metabolismo , Factor de Crecimiento Transformador beta1/genéticaRESUMEN
The authors described three groups of patients after acute poisonings. In the first group were 60 patients after carbon tetrachioride poisoning, the second group consisted of 81 patients after mushroom poisoning and 20 patients after ethylene glycol poisoning were in the third group. Besides two patients with rare poisonings after potassium dichromate and after paraquat poisoning were analysed. All groups of patients with the kidney damage were presented from the diagnostic, differential diagnostic, conservative, ntra- and extracorporeal elimination treatment point of view. In the group of patients suffering from acute carbon tetrachloride poisoning and with acute renal failure following therapy was used: conservative treatment, exchange blood transfusion--in 4 patients in hepatic coma, renal replacement therapy (peritoneal dialysis, haemodialysis, plasmapheresis). From the total number of 60 patients 58 survived and 2 patients died in liver coma. Survival of patients after mushroom poisoning depended on amount of oral use of mushroom (Amanita phalloides), on early admission in dialysis centre and on early beginning of renal replacement therapy within 24 hr after acute poisoning. Twenty four patients from 81 patients of this group died. Main clinical signs of ethylene glycol poisoning were various neurological symptoms (cramps, hemiparesis, coma), severe metabolic acidosis (pH = 7.06 +/- 0.14), leucocytosis (26.4 +/- 5.5x 10(9)/L) and the signs of acute toxic hepatitis and of acute renal failure. Calcium oxalic crystals in urine were present in 17 patients and leucocytosis was observed in every patient. In the first 4 patients we administered intravenously ethylalcohol as an antidotum and later in other patients we used ethylalcohol in dialysis solution. The concentration of ethylalcohol in dialysis solution was 100 mg%. Severe metabolic acidosis improved in 17 patients using bicarbonate haemodialysis and 3 patients died before the possibility to use bicarbonate haemodialysis. Eighty-four hours after acute potassium dichromate poisoning and 24 hours after exchange blood transfusion during haemodialysis a 41-year old man died in haemorhagic shock, which developed after the extensive chemical burns of mucous membrane of gastrointestinal tract caused by this poison. Our patient after paraquat poisoning was treated by repeated charcoal haemoperfusion and haemodialysis. Despite of that therapy the patient died in severe respiratory insufficiency.
Asunto(s)
Lesión Renal Aguda/terapia , Intoxicación por Tetracloruro de Carbono/terapia , Sobredosis de Droga/terapia , Glicol de Etileno/envenenamiento , Intoxicación por Setas/terapia , Terapia de Reemplazo Renal , Lesión Renal Aguda/etiología , Adulto , Amanita , Transfusión Sanguínea , Quemaduras Químicas/etiología , Intoxicación por Tetracloruro de Carbono/complicaciones , Intoxicación por Tetracloruro de Carbono/mortalidad , Carbón Orgánico/uso terapéutico , Sobredosis de Droga/complicaciones , Sobredosis de Droga/mortalidad , Tracto Gastrointestinal/efectos de los fármacos , Tracto Gastrointestinal/lesiones , Humanos , Masculino , Intoxicación por Setas/complicaciones , Intoxicación por Setas/mortalidad , Paraquat/envenenamiento , Dicromato de Potasio/envenenamiento , Diálisis Renal , Choque Hemorrágico/etiología , Tasa de SupervivenciaRESUMEN
BACKGROUND: Opioids have been shown to attenuate ischaemia-reperfusion injury (IRI) in a number of organs. We evaluated the effect of morphine pretreatment on IRI in both normal and cirrhotic rat liver. METHODS: Morphine was administered either i.v. or intrathecally (i.t.) 10 min before initiating 1 h of ischaemia followed by 6 h reperfusion in normal rat liver. Hepatic injury was assessed histologically using Suzuki's criteria. These manoeuvres were repeated using the optimal dose of morphine after administration of naloxone methiodide and wortmannin. Serum levels of transaminases were measured, and expression of phosphorylated Akt, Jak2, and STAT3 were assessed by immunoblotting. Similar procedures were repeated on rats with carbon tetrachloride-induced liver cirrhosis, and the levels of phosphorylated protein kinase C (PKC), haem oxygenase-1 (HO-1), and inducible nitric oxide synthase (iNOS) were also evaluated, as these proteins have beneficial effects during IRI. RESULTS: Morphine pretreatment at 100 µg kg(-1) (i.v.) or 10 µg (i.t.) reduced necrosis, apoptosis, and serum transaminase levels, and increased phosphorylated Akt and STAT3 but not JAK2 expression in normal liver. These changes were reversed by prior administration of naloxone methiodide and wortmannin. Although morphine preconditioning was also protective in cirrhotic liver, STAT3 and JAK2 phosphorylation status was unchanged. There was, however, increased expression of phosphorylated PKC and HO-1, and a reduction in iNOS. CONCLUSIONS: Morphine preconditioning protects against IRI in both normal and cirrhotic rat liver. This involves opioid receptors, phosphatidylinositol-3-kinase, and Akt. The downstream pathways involved are different for cirrhotic liver, with preliminary evidence suggesting involvement of HO-1.
Asunto(s)
Analgésicos Opioides/uso terapéutico , Cirrosis Hepática Experimental/complicaciones , Hígado/lesiones , Morfina/uso terapéutico , Daño por Reperfusión/prevención & control , Alanina Transaminasa/sangre , Analgésicos Opioides/administración & dosificación , Animales , Apoptosis/efectos de los fármacos , Aspartato Aminotransferasas/sangre , Western Blotting , Intoxicación por Tetracloruro de Carbono/terapia , Cateterismo , Hemo-Oxigenasa 1/biosíntesis , Etiquetado Corte-Fin in Situ , Inyecciones Intravenosas , Inyecciones Espinales , Precondicionamiento Isquémico , Hígado/patología , Cirrosis Hepática Experimental/inducido químicamente , Cirrosis Hepática Experimental/patología , Masculino , Morfina/administración & dosificación , Ratas , Ratas Sprague-Dawley , Daño por Reperfusión/patología , Transducción de Señal/efectos de los fármacosRESUMEN
Niemann-Pick disease (NPD) is a lysosomal storage disease caused by the loss of acid sphingomyelinase (ASMase) that features neurodegeneration and liver disease. Because ASMase-knock-out mice models NPD and our previous findings revealed that ASMase activates cathepsins B/D (CtsB/D), our aim was to investigate the expression and processing of CtsB/D in hepatic stellate cells (HSCs) from ASMase-null mice and their role in liver fibrosis. Surprisingly, HSCs from ASMase-knock-out mice exhibit increased basal level and activity of CtsB as well as its in vitro processing in culture, paralleling the enhanced expression of fibrogenic markers α-smooth muscle actin (α-SMA), TGF-ß, and pro-collagen-α1(I) (Col1A1). Moreover, pharmacological inhibition of CtsB blunted the expression of α-SMA and Col1A1 and proliferation of HSCs from ASMase-knock-out mice. Consistent with the enhanced activation of CtsB in HSCs from ASMase-null mice, the in vivo liver fibrosis induced by chronic treatment with CCl(4) increased in ASMase-null compared with wild-type mice, an effect that was reduced upon CtsB inhibition. In addition to liver, the enhanced proteolytic processing of CtsB was also observed in brain and lung of ASMase-knock-out mice, suggesting that the overexpression of CtsB may underlie the phenotype of NPD. Thus, these findings reveal a functional relationship between ASMase and CtsB and that the ablation of ASMase leads to the enhanced processing and activation of CtsB. Therefore, targeting CtsB may be of relevance in the treatment of liver fibrosis in patients with NPD.
Asunto(s)
Catepsina B/metabolismo , Cirrosis Hepática/enzimología , Enfermedades de Niemann-Pick/enzimología , Esfingomielina Fosfodiesterasa/metabolismo , Animales , Biomarcadores/metabolismo , Tetracloruro de Carbono/toxicidad , Intoxicación por Tetracloruro de Carbono/genética , Intoxicación por Tetracloruro de Carbono/metabolismo , Intoxicación por Tetracloruro de Carbono/patología , Intoxicación por Tetracloruro de Carbono/terapia , Catepsina B/genética , Catepsina D/genética , Catepsina D/metabolismo , Proliferación Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Humanos , Cirrosis Hepática/inducido químicamente , Cirrosis Hepática/genética , Cirrosis Hepática/patología , Cirrosis Hepática/terapia , Ratones , Ratones Noqueados , Enfermedades de Niemann-Pick/inducido químicamente , Enfermedades de Niemann-Pick/genética , Enfermedades de Niemann-Pick/patología , Enfermedades de Niemann-Pick/terapia , Esfingomielina Fosfodiesterasa/genéticaRESUMEN
BACKGROUND: Human amniotic membrane-derived mesenchymal stem cells (hAMCs) have the potential to reduce heart and lung fibrosis, but whether could reduce liver fibrosis remains largely unknown. METHODOLOGY/PRINCIPAL FINDINGS: Hepatic cirrhosis model was established by infusion of CCl4 (1 ml/kg body weight twice a week for 8 weeks) in immunocompetent C57Bl/6J mice. hAMCs, isolated from term delivered placenta, were infused into the spleen at 4 weeks after mice were challenged with CCl4. Control mice received only saline infusion. Animals were sacrificed at 4 weeks post-transplantation. Blood analysis was performed to evaluate alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Histological analysis of the livers for fibrosis, hepatic stellate cells activation, hepatocyte apoptosis, proliferation and senescence were performed. The donor cell engraftment was assessed using immunofluorescence and polymerase chain reaction. The areas of hepatic fibrosis were reduced (6.2%±2.1 vs. control 9.6%±1.7, p<0.05) and liver function parameters (ALT 539.6±545.1 U/dl, AST 589.7±342.8 U/dl,vs. control ALT 139.1±138.3 U/dl, p<0.05 and AST 212.3±110.7 U/dl, p<0.01) were markedly ameliorated in the hAMCs group compared to control group. The transplantation of hAMCs into liver-fibrotic mice suppressed activation of hepatic stellate cells, decreased hepatocyte apoptosis and promoted liver regeneration. More interesting, hepatocyte senescence was depressed significantly in hAMCs group compared to control group. Immunofluorescence and polymerase chain reaction revealed that hAMCs engraftment into host livers and expressed the hepatocyte-specific markers, human albumin and α-fetoproteinran. CONCLUSIONS/SIGNIFICANCE: The transplantation of hAMCs significantly decreased the fibrosis formation and progression of CCl4-induced cirrhosis, providing a new approach for the treatment of fibrotic liver disease.
Asunto(s)
Amnios/citología , Intoxicación por Tetracloruro de Carbono/terapia , Cirrosis Hepática Experimental/terapia , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/fisiología , Animales , Tetracloruro de Carbono , Intoxicación por Tetracloruro de Carbono/patología , Células Cultivadas , Femenino , Hepatocitos/patología , Hepatocitos/fisiología , Humanos , Cirrosis Hepática Experimental/inducido químicamente , Cirrosis Hepática Experimental/patología , Regeneración Hepática/fisiología , Células Madre Mesenquimatosas/citología , Ratones , Ratones Endogámicos C57BLRESUMEN
We have developed an in vivo mouse model [the green fluorescent protein (GFP) / carbon tetrachloride (CCl(4)) model] and reported that infused GFP-positive bone marrow cells administered via a tail vein efficiently repopulated cirrhotic liver tissue under conditions of persistent liver damage induced by CCl(4). Moreover, bone marrow cells infused into the liver improved liver function and ameliorated liver fibrosis with higher expression of matrix metalloproteinase 9 (MMP-9), consistent with improved survival rate. Based on these findings, we started a multicenter clinical trial of autologous bone marrow cell infusion (ABMi) therapy for decompensated liver cirrhosis patients and demonstrated the efficacy of this approach without unexpected complications. However, this therapy involves bone marrow aspiration under general anesthesia and is not indicated for patients for whom general anesthesia is difficult. We therefore aimed to develop a new liver regeneration therapy in which cells having a curative effect on liver cirrhosis are isolated and cultured from a small amount of autologous bone marrow aspirated under local anesthesia and infused back into the same subject. Herein, we present results for the GFP/CCl(4) model and ABMi therapy and future prospects for a new liver regeneration therapy.
Asunto(s)
Trasplante de Médula Ósea , Cirrosis Hepática/terapia , Hepatopatías/terapia , Regeneración Hepática , Hígado/fisiopatología , Hígado/cirugía , Animales , Intoxicación por Tetracloruro de Carbono/metabolismo , Intoxicación por Tetracloruro de Carbono/terapia , Ensayos Clínicos como Asunto , Modelos Animales de Enfermedad , Femenino , Proteínas Fluorescentes Verdes/metabolismo , Proteínas Fluorescentes Verdes/farmacología , Humanos , Cirrosis Hepática/inducido químicamente , Cirrosis Hepática/cirugía , Hepatopatías/cirugía , Masculino , Ratones , Estudios Multicéntricos como AsuntoRESUMEN
This experiment was preformed on an animal model of CCl4 hepatitis using 320 Wistar rats. The animals underwent combined effect of saprogel and a magnetic field during different seasons. It was shown that non-medicamentous treatment of experimental CCl4 hepatitis had the most pronounced beneficial effect on the synthetic liver function in the winter time. An appreciable suppression of the hepatic function was documented in the spring.
Asunto(s)
Balneología , Intoxicación por Tetracloruro de Carbono , Hepatitis Animal , Hígado/metabolismo , Magnetoterapia , Ácidos Nucleicos/biosíntesis , Estaciones del Año , Animales , Intoxicación por Tetracloruro de Carbono/metabolismo , Intoxicación por Tetracloruro de Carbono/terapia , Modelos Animales de Enfermedad , Hepatitis Animal/metabolismo , Hepatitis Animal/terapia , Masculino , Ratas , Ratas WistarRESUMEN
Bone marrow stem cells nullify acquired and non-acquired diseases of liver through multiple strategies including antiinflammation. However, little is known about the in vivo mechanism of immunomodulation by stem cells in mediating liver cirrhosis. Mesenchymal stem cells (MSC) or hematopoietic stem cells (HSC) isolated from bone marrow of male mice were transplanted into female mice with acute liver inflammation. Serum levels of liver proteins and aminotransferase as well as hepatic antioxidant enzymes were estimated. Immunostaining for the expression of tumor necrosis factor alpha (TNF-alpha), interleukin 6 (IL-6), alpha smooth muscle actin (alpha-SMA) and type I collagen proteins was carried out and the expression of these mRNAs was also studied. After post-transplantation, the levels of serum albumin and aminotransferases became normal and the levels of antioxidants were significantly high in the MSC treated mice compared to HSC and control mice. Necrotic cells and invasion of neutrophils were not observed in histological sections of liver of MSC treated mice. Immunostaining showed that IL-6 and TNF-alpha were not expressed in the MSC treated mice when compared to the control and HSC treated mice. alpha-SMA representing activated myofibroblasts and type I collagen were not expressed in MSC treated group. These inflammatory and fibrogenic results were further confirmed by reverse transcription-polymerase chain reaction (RT-PCR). The acute inflammation ended with the formation of fibrosis in the HSC and control groups by the uncontrolled immunoreactions. Protection mechanism of MSC therapy against injury and fibrosis in the liver occurs by the suppression of inflammation. Our findings suggest that bone marrow MSC are capable of alleviating the immunoreactions leading to the fibrosis in the liver.
Asunto(s)
Intoxicación por Tetracloruro de Carbono/terapia , Enfermedad Hepática Inducida por Sustancias y Drogas/terapia , Fibrosis/terapia , Trasplante de Células Madre Hematopoyéticas , Inflamación/terapia , Trasplante de Células Madre Mesenquimatosas , Adipocitos/fisiología , Animales , Antioxidantes/metabolismo , Diferenciación Celular , Separación Celular , Fibrosis/inducido químicamente , Inflamación/inducido químicamente , Peroxidación de Lípido , Hígado/enzimología , Pruebas de Función Hepática , Ratones , Ratones Endogámicos BALB C , Osteocitos/fisiología , Peroxidasa/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Transplantation of hepatocytes or bone marrow-derived cells has been shown to ameliorate liver fibrosis in animal models, but no direct comparison of relative efficiency has been made. The aim of this study was to compare the efficiency of a bone marrow-derived clonal mesenchymal stem cell line established by us (rBM25/S3) with that of its adipogenic or hepatogenic differentiation derivative for suppression of rat liver fibrosis. After induction of differentiation of rBM25/S3 cells into adipogenic or hepatogenic cells in culture, we intrasplenically transplanted the three types of cells into rats (3 x 10(7) cells/rat) before and 4 weeks after initiation of carbon tetrachloride treatment (1 ml/kg body weight twice a week for 8 weeks) to induce liver fibrosis. Undifferentiated rBM25/S3 cells were the most effective for suppression of liver fibrosis, followed by the adipogenic cells and hepatogenic cells. Expression levels of MMP-2 and MMP-9 were also highest in undifferentiated rBM25/S3 cells. These results indicate that bone marrow-derived clonal mesenchymal stem cell lines are useful for further mechanistic studies on cell-mediated suppression of liver fibrosis and that such cell lines will provide information on an appropriate cell source for transplantation therapy for cirrhosis.
Asunto(s)
Células de la Médula Ósea/citología , Cirrosis Hepática Experimental/terapia , Trasplante de Células Madre Mesenquimatosas , Animales , Tetracloruro de Carbono/toxicidad , Intoxicación por Tetracloruro de Carbono/terapia , Línea Celular Tumoral , Células Cultivadas , Modelos Animales de Enfermedad , Cirrosis Hepática Experimental/inducido químicamente , Cirrosis Hepática Experimental/prevención & control , Masculino , Ratas , Ratas Sprague-Dawley , Células Tumorales CultivadasRESUMEN
Free radicals cause cell injury, when they are generated in excess or when the antioxidant defense is impaired. Carbon tetrachloride (CCl4) is used as a model for liver injury. In this study antioxidant activity of ethanol extract of A. fertilisima (EEA) was investigated using CCl4 intoxicated rat liver as the experimental model. Oral administration of EEA at a dose of 100 mg/kg body weight, for 14 consecutive days, the rate of the production of antioxidant enzymes like super oxide dismutase, catalase, glutathione peroxidase and glutathione transferase in rats compared to the CCl4 treated group without any supporting treatment. Liver damage is detected by the measurement of the activities of serum enzymes like aspartate aminotransferase, alanine aminotransferase, gamma glutamyl transpeptidase and alkaline phosphatase which were released in to the blood from damaged cells. The normalization of these enzymes levels was observed in rats treated with EEA (100 mg/kg body weight) by reducing the leakage of the above enzymes in to the blood. The findings provide a rationale for further studies on isolation of active principles and its pharmacological evaluation. Protection offered by silymarin (standard reference drug) seemed relatively greater.