RESUMEN
The root of Codonopis bulleynana Forest ex Diels (cbFeD), a tonic food widely used in Yunnan Province of China, was found to have a wide range of pharmacological effects. The present study was designed to investigate the anti-fibrotic effect of water extracts of cbFeD in chronic liver injury mice model induced by carbon tetrachloride (CCl4) and to explore its underlying mechanisms. Phytochemical analysis revealed multiple components were present in the water extract of cbFeD and the compounds were mostly enriched in organic acid and its derivatives, flavone, amino acid derivatives, nucleotide and its derivatives, carbohydrates etc. Treatment with cbFeD significantly attenuated liver injury and fibrosis in CCl4-administered mice evidenced by improved liver histology, ameliorated apoptosis of hepatocytes, and decreased transaminase levels in the serum. Decreased activities of superoxide dismutase (SOD) and catalase (CAT) were markedly reversed upon treatment with cbFeD while levels of malondialdehyde (MDA) and glutathione (GSH) were significantly restored towards normal values. cbFeD also suppressed intrahepatic inflammatory cell infiltration and Kupffer cell activation. Furthermore, our study revealed an inhibitory effect of cbFeD on hepatic stellate cells (HSCs) activation both in vitro and in vivo. In conclusion, cbFeD could exert a protective role against liver fibrosis in mice model induced by CCl4 that is comparable to the positive control silymarin and might be developed into a promising anti-fibrotic drug.
Asunto(s)
Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , Codonopsis , Células Estrelladas Hepáticas/efectos de los fármacos , Cirrosis Hepática Experimental/prevención & control , Hígado/efectos de los fármacos , Extractos Vegetales/farmacología , Animales , Apoptosis/efectos de los fármacos , Tetracloruro de Carbono , Línea Celular , Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Quimiotaxis de Leucocito/efectos de los fármacos , Codonopsis/química , Células Estrelladas Hepáticas/metabolismo , Células Estrelladas Hepáticas/patología , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , Hepatocitos/patología , Humanos , Mediadores de Inflamación/metabolismo , Macrófagos del Hígado/efectos de los fármacos , Macrófagos del Hígado/metabolismo , Macrófagos del Hígado/patología , Hígado/metabolismo , Hígado/patología , Cirrosis Hepática Experimental/inducido químicamente , Cirrosis Hepática Experimental/metabolismo , Cirrosis Hepática Experimental/patología , Activación de Macrófagos/efectos de los fármacos , Masculino , Ratones , Estrés Oxidativo/efectos de los fármacos , Extractos Vegetales/aislamiento & purificación , Raíces de PlantasRESUMEN
BACKGROUND Kupffer cells and natural killer (NK) cells has been identified as contributing factors in the pathogenesis of hepatitis, but the detailed mechanism of these cell types in the pathogenesis of primary biliary cholangitis (PBC) is poorly understood. MATERIAL AND METHODS In this study, polyinosinic: polycytidylic acid (poly I: C), 2-octynoic acid-bovine serum albumin (2OA-BSA) and Freund's adjuvant (FA) were injected to establish a murine PBC model, from which NK cells and Kupffer cells were extracted and isolated. The cells were then co-cultivated in a designed culture system, and then NK group 2, member D (NKG2D), retinoic acid early inducible-1 (RAE-1), F4/80, and cytokine expression levels were detected. RESULTS The results showed close crosstalk between Kupffer cells and NK cells. PBC mice showed increased surface RAE-1 protein expression and Kupffer cell cytokine secretion, which subsequently activated NK cell-mediated target cell killing via NKG2D/RAE-1 recognition, and increased inflammation. NK cell-derived interferon-γ (IFN-γ) and Kupffer cell-derived tumor necrosis factor alpha (TNF-alpha) were found to synergistically regulate inflammation. Moreover, interleukin (IL)-12 and IL-10 improved the crosstalk between NK cells and Kupffer cells. CONCLUSIONS Our ï¬ndings in mice are the first to suggest the involvement of the NKG2D/RAE-1 interaction and cytokines in the synergistic effects of NK and Kupffer cells in PBC.
Asunto(s)
Células Asesinas Naturales/metabolismo , Macrófagos del Hígado/metabolismo , Cirrosis Hepática Biliar/metabolismo , Animales , Citocinas/metabolismo , Modelos Animales de Enfermedad , Femenino , Interferón gamma/metabolismo , Interleucina-12/metabolismo , Células Asesinas Naturales/patología , Macrófagos del Hígado/patología , Cirrosis Hepática Biliar/fisiopatología , Ratones , Ratones Endogámicos C57BL , Subfamilia K de Receptores Similares a Lectina de Células NK/metabolismo , Proteínas Asociadas a Matriz Nuclear/metabolismo , Proteínas de Transporte Nucleocitoplasmático/metabolismoRESUMEN
Selenizing astragalus polysaccharides-3 (sAPS3) was prepared by nitric acid-sodium selenite method. The effects of sAPS3 on carbon tetrachloride (CCl4) induced hepatocellular necrosis, and its underlying mechanisms were studied in male Wistar rats. Hepatic damage was induced by intraperitoneal injection of CCl4 twice a week, for 3 weeks. Meanwhile, the rats in addition to CCl4 were also exposed to sodium selenite (SS), astragalus polysaccharides (APS), SS + APS or sAPS3, in parallel by oral gavage once a day for 3 weeks. At the end of 3 weeks, blood and liver tissue were taken. Serum was collected to test the levels of alanine aminotransferase, aspartate aminotransferase and antioxidant status parameters. Liver tissue was collected for histopathological examination and determination of messenger RNA (mRNA) expression levels of CD68, TNF-α, IL-1ß and ATG7 followed by the measurements of CD68, IL-1ß and LC3II by immunohistochemistry assay (IHC), or TNF-α by immunofluorescence assay (IFA). The results showed that sAPS3 effectively ameliorated CCl4 induced hepatocellular necrosis and inflammation and significantly decreased the levels of aspartate aminotransferase, alanine aminotransferase, malondialdehyde and the expression levels of Kupffer cells (KCs)-specific biomarker CD68 and proinflammatory cytokines produced by activated KCs such as IL-1ß and TNF-α (P < 0.01). While increasing the levels of total antioxidant capacity, glutathione, glutathione peroxidase and superoxide dismutase (P < 0.05) and reduced the expression levels of a key regulator of autophagy in KCs ATG7 or LC3II (P < 0.05). These findings indicate that sAPS3 could ameliorate CCl4-induced hepatocellular necrosis by inactivation of Kupffer cells and its activity may be superior to the application of selenium, APS or combination of selenium with APS.
Asunto(s)
Astragalus propinquus/química , Macrófagos del Hígado/efectos de los fármacos , Hígado/patología , Polisacáridos/farmacología , Selenio/farmacología , Animales , Antígenos CD/metabolismo , Antígenos de Diferenciación Mielomonocítica/metabolismo , Tetracloruro de Carbono/toxicidad , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , Citocinas/metabolismo , Relación Dosis-Respuesta a Droga , Enzimas/sangre , Glutatión/metabolismo , Macrófagos del Hígado/patología , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Malondialdehído/metabolismo , Necrosis/inducido químicamente , Necrosis/prevención & control , Polisacáridos/administración & dosificación , Polisacáridos/química , Ratas Wistar , Selenio/administración & dosificaciónRESUMEN
Agastache rugosa (A. rugosa, Labiatae), a perennial herb spread throughout Korean fields, is widely consumed as a wild edible vegetable and is used in folk medicine. This study examined the hepatoprotective mechanisms of ß-caryophyllene (BCP), a major bicyclic sesquiterpene of A. rugosa, against D-galactosamine (GalN) and lipopolysaccharide (LPS)-induced hepatic failure. Mice were given an intraperitoneal injection of BCP (50, 100 and 200 mg/kg) 1 h before GalN (800 mg/kg)/LPS (40 µg/kg) injection and were killed 1 h or 6 h after GalN/LPS injection. GalN/LPS markedly increased mortality and serum aminotransferase activity, both of which were attenuated by BCP. BCP also attenuated increases in serum tumor necrosis factor-α, interleukin 6, and high-mobility group protein B1 levels by GalN/LPS. GalN/LPS significantly increased toll-like receptor (TLR) 4 and receptor for advanced glycation end products (RAGE) protein expression, extracellular signal-related kinase, p38 and c-Jun N-terminal kinase phosphorylation, nuclear factor κB (NF-κB), early growth response protein-1, and macrophage inflammatory protein-2 protein expression. These increases were attenuated by BCP. Furthermore, BCP suppressed increased TLR4 and RAGE protein expression and proinflammatory cytokines production in LPS-treated isolated Kupffer cells. Our findings suggest that BCP protects against GalN/LPS-induced liver injury through down-regulation of the TLR4 and RAGE signaling.
Asunto(s)
Antiinflamatorios/farmacología , Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , Galactosamina , Lipopolisacáridos , Fallo Hepático/prevención & control , Hígado/efectos de los fármacos , Receptor para Productos Finales de Glicación Avanzada/efectos de los fármacos , Sesquiterpenos/farmacología , Receptor Toll-Like 4/efectos de los fármacos , Animales , Células Cultivadas , Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Citocinas/metabolismo , Citoprotección , Modelos Animales de Enfermedad , Mediadores de Inflamación/metabolismo , Macrófagos del Hígado/efectos de los fármacos , Macrófagos del Hígado/metabolismo , Macrófagos del Hígado/patología , Hígado/metabolismo , Hígado/patología , Fallo Hepático/metabolismo , Fallo Hepático/patología , Masculino , Ratones Endogámicos ICR , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Sesquiterpenos Policíclicos , Receptor para Productos Finales de Glicación Avanzada/metabolismo , Transducción de Señal/efectos de los fármacos , Factores de Tiempo , Receptor Toll-Like 4/metabolismoRESUMEN
This study investigated the effects of low-level laser therapy (LLLT) in the liver function, structure and inflammation in a experimental model of carbon tetrachloride (CCl(4))-induced liver cirrhosis. Wistar rats were divided into Control, LLLT, CCl(4) and CCl(4) +LLLT groups. CCl(4) groups received CCl(4) (0.4 g kg(-1); i.p.), three times a week, for 12 weeks. A 830 nm LLLT was performed with a continuous wave, 35 mW, 2.5 J cm(-2) per point, applied to four points of the liver (right and left upper and lower extremities, in the four lobes of the liver) for 2 weeks. Liver structure and inflammation (cirrhotic areas, collagen deposition, inflammation, density of Kupffer and hepatic stellate cells) and function (aspartate aminotransferase, alkaline phosphatase, gamma glutamyltransferase, lactate dehydrogenase, total proteins and globulins) were evaluated. LLLT significantly reduced CCl(4)-increased aspartate aminotransferase (P < 0.001), alkaline phosphatase (P < 0.001), gamma-glutamyl transferase (P < 0.001) and lactate dehydrogenase (P < 0.01) activity, as well as total proteins (P < 0.05) and globulins (P < 0.01). LLLT also reduced the number of cirrhotic areas, the collagen accumulation and the hepatic inflammatory infiltrate. Of note, LLLT reduced CCl(4)-increased number of Kupffer cells (P < 0.05) and hepatic stellate cells (P < 0.05). We conclude that LLLT presents beneficial effects on liver function and structure in an experimental model of CCl(4)-induced cirrhosis.
Asunto(s)
Células Estrelladas Hepáticas/efectos de la radiación , Macrófagos del Hígado/efectos de la radiación , Cirrosis Hepática/radioterapia , Terapia por Luz de Baja Intensidad , Alanina Transaminasa/metabolismo , Fosfatasa Alcalina/metabolismo , Animales , Tetracloruro de Carbono , Recuento de Células , Colágeno/metabolismo , Regulación hacia Abajo , Células Estrelladas Hepáticas/metabolismo , Células Estrelladas Hepáticas/patología , Macrófagos del Hígado/metabolismo , Macrófagos del Hígado/patología , L-Lactato Deshidrogenasa/metabolismo , Cirrosis Hepática/inducido químicamente , Cirrosis Hepática/metabolismo , Cirrosis Hepática/patología , Pruebas de Función Hepática , Masculino , Ratas , Ratas Wistar , gamma-Glutamiltransferasa/metabolismoRESUMEN
BACKGROUND & AIMS: Progression from steatosis to steatohepatitic lesions is hypothesized to require a second hit. These lesions have been associated with increased oxidative stress, often ascribed to high levels of leptin and other proinflammatory mediators. Here we have examined the role of leptin in inducing oxidative stress and Kupffer cell activation in CCl4-mediated steatohepatitic lesions of obese mice. METHODS: Male C57BL/6 mice fed with a high-fat diet (60%kcal) at 16 weeks were administered CCl4 to induce steatohepatitic lesions. Approaches included use of immuno-spin trapping for measuring free radical stress, gene-deficient mice for leptin, p47 phox, iNOS and adoptive transfer of leptin primed macrophages in vivo. RESULTS: Diet-induced obese (DIO) mice, treated with CCl4 increased serum leptin levels. Oxidative stress was significantly elevated in the DIO mouse liver, but not in ob/ob mice, or in DIO mice treated with leptin antibody. In ob/ob mice, leptin supplementation restored markers of free radical generation. Markers of free radical formation were significantly decreased by the peroxynitrite decomposition catalyst FeTPPS, the iNOS inhibitor 1400W, the NADPH oxidase inhibitor apocynin, or in iNOS or p47 phox-deficient mice. These results correlated with the decreased expression of TNF-alpha and MCP-1. Kupffer cell depletion eliminated oxidative stress and inflammation, whereas in macrophage-depleted mice, the adoptive transfer of leptin-primed macrophages significantly restored inflammation. CONCLUSIONS: These results, for the first time, suggest that leptin action in macrophages of the steatotic liver, through induction of iNOS and NADPH oxidase, causes peroxynitrite-mediated oxidative stress thus activating Kupffer cells.
Asunto(s)
Hígado Graso/metabolismo , Macrófagos del Hígado/metabolismo , Leptina/metabolismo , Estrés Oxidativo , Animales , Citocinas/metabolismo , Modelos Animales de Enfermedad , Hígado Graso/etiología , Mediadores de Inflamación/metabolismo , Macrófagos del Hígado/patología , Masculino , Ratones , Ratones Endogámicos C57BL , NADPH Oxidasas/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Enfermedad del Hígado Graso no Alcohólico , Obesidad/complicaciones , Ácido Peroxinitroso/metabolismoRESUMEN
Alcoholic liver disease (ALD) is characterized by steatosis and upregulation of proinflammatory cytokines, including IL-1ß. IL-1ß, type I IL-1 receptor (IL-1R1), and IL-1 receptor antagonist (IL-1Ra) are all important regulators of the IL-1 signaling complex, which plays a role in inflammation. Furthermore, IL-1ß maturation is dependent on caspase-1 (Casp-1). Using IL-1Ra-treated mice as well as 3 mouse models deficient in regulators of IL-1ß activation (Casp-1 and ASC) or signaling (IL-1R1), we found that IL-1ß signaling is required for the development of alcohol-induced liver steatosis, inflammation, and injury. Increased IL-1ß was due to upregulation of Casp-1 activity and inflammasome activation. The pathogenic role of IL-1 signaling in ALD was attributable to the activation of the inflammasome in BM-derived Kupffer cells. Importantly, in vivo intervention with a recombinant IL-1Ra blocked IL-1 signaling and markedly attenuated alcohol-induced liver inflammation, steatosis, and damage. Furthermore, physiological doses of IL-1ß induced steatosis, increased the inflammatory and prosteatotic chemokine MCP-1 in hepatocytes, and augmented TLR4-dependent upregulation of inflammatory signaling in macrophages. In conclusion, we demonstrated that Casp-1-dependent upregulation of IL-1ß and signaling mediated by IL-1R1 are crucial in ALD pathogenesis. Our findings suggest a potential role of IL-1R1 inhibition in the treatment of ALD.
Asunto(s)
Hígado Graso Alcohólico/tratamiento farmacológico , Inflamasomas/fisiología , Proteína Antagonista del Receptor de Interleucina 1/uso terapéutico , Animales , Proteínas Portadoras/biosíntesis , Proteínas Portadoras/genética , Caspasa 1/fisiología , Quimiocina CCL2/biosíntesis , Quimiocina CCL2/genética , Evaluación Preclínica de Medicamentos , Etanol/toxicidad , Hígado Graso Alcohólico/etiología , Femenino , Hepatocitos/metabolismo , Interleucina-1alfa/biosíntesis , Interleucina-1alfa/sangre , Interleucina-1beta/biosíntesis , Interleucina-1beta/sangre , Interleucina-1beta/fisiología , Interleucina-1beta/toxicidad , Macrófagos del Hígado/patología , Cirrosis Hepática/etiología , Cirrosis Hepática/prevención & control , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Proteína con Dominio Pirina 3 de la Familia NLR , Receptores Tipo I de Interleucina-1/deficiencia , Receptores Tipo I de Interleucina-1/fisiología , Proteínas Recombinantes/uso terapéutico , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 4/fisiología , Regulación hacia Arriba/efectos de los fármacosRESUMEN
OBJECTIVE: To observe the effects of different therapeutic methods and the recipes of Chinese medicine (CM) on the activation of c-Jun N-terminal kinase (JNK) in Kupffer cells of rats with fatty liver disease and to explore the mechanisms of these therapeutic methods. METHODS: By using a random number table, 98 rats were randomly divided into 7 groups: control group, model group, and 5 treatment groups, including soothing Liver (Gan) recipe group, invigorating Spleen (Pi) recipe group, dispelling dampness recipe group, promoting blood recipe group, and complex recipe group. Rats in the control group were fed with normal food and distilled water by gastric perfusion, while rats in the model group were fed with high-fat food and distilled spirits by gastric perfusion. Rats in the 5 treatment groups were fed with high-fat food and corresponding recipes by gastric perfusion. Twelve weeks later, all rats were sacrificed and liver tissues were stained for pathohistological observation. Kupffer cells were isolated from livers of rats to evaluate JNK and phospho-JNK expressions by Western blotting. RESULTS: The grade of hepatic steatosis was higher in the model group than the control group (P<0.05). Compared with the model group, the grade of fatty degeneration in soothing Liver recipe group and invigorating Spleen recipe group were significantly ameliorated (P<0.05). Expressions of JNK and phospho-JNK in Kupffer cells were significantly higher in the model group than those in the control group (P<0.05, P<0.01). Compared with the model group, expressions of JNK in all treatment groups decreased, especially in invigorating Spleen recipe group and promoting blood recipe group (P<0.05). Compared with the model group, expressions of phospho-JNK in all treatment groups declined significantly (P<0.01), especially in soothing Live recipe group and invigorating Spleen recipe group. CONCLUSIONS: The high expressions of JNK and phospho-JNK in Kupffer cells might play an important role in the pathogenesis of fatty liver disease in rats. The recipes of CM, especially invigorating Spleen recipe and soothing Liver recipe, might protect liver against injury by reducing the total JNK protein content and inhibiting the activation of JNK protein in Kupffer cells of fatty liver model rats, which showed beneficial effects on fatty liver disease.
Asunto(s)
Medicamentos Herbarios Chinos/farmacología , Hígado Graso/enzimología , Hígado Graso/terapia , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Macrófagos del Hígado/enzimología , Macrófagos del Hígado/patología , Animales , Supervivencia Celular/efectos de los fármacos , Activación Enzimática/efectos de los fármacos , Hígado Graso/patología , Hepatocitos/efectos de los fármacos , Hepatocitos/enzimología , Hepatocitos/patología , Macrófagos del Hígado/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/enzimología , Hígado/patología , Fosforilación/efectos de los fármacos , Ratas , Ratas Sprague-DawleyRESUMEN
Activation of myofibroblasts (MF) and extracellular matrix (ECM) deposition predispose the expansion and differentiation of liver progenitor cells (LPC) during chronic liver injury. Because Kupffer cells (KC) are active modulators of tissue response and fibrosis, we analyzed their role in a model of LPC proliferation. A choline-deficient diet, supplemented by ethionine (CDE) was administrated to C57Bl/6J mice that were depleted of KC by repeated injections of clodronate (CLO) and compared to PBS-injected mice. On CDE, massive KC activation was observed in the PBS group, but this was blunted in CLO-treated mice. The depletion of KC did not influence LPC proliferation but reduced their invasive behavior. Instead of being found far into the parenchyma, as was found in the PBS group (mean distance from portal vein: 209 µm), LPC of CLO mice remained closer to the portal area (138 µm), forming aggregates and phenotypically resembling cells of biliary lineage. Notably, removal of KC was also associated with a significant decrease in amount of MF and ECM and in the expression of profibrotic factors. Thus, besides ECM and MF, KC are also a significant component of the microenvironmental changes preceding LPC expansion. Depletion of KC may limit the LPC parenchymal invasion through a deficiency in chemoattracting factors, reduced activation of MF, and/or a paucity of the ECM framework necessary for cell motility.
Asunto(s)
Movimiento Celular , Macrófagos del Hígado/patología , Hepatopatías/patología , Hígado/patología , Células Madre/patología , Animales , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Colina/farmacología , Citocinas/metabolismo , Dieta , Modelos Animales de Enfermedad , Etionina/farmacología , Matriz Extracelular/efectos de los fármacos , Matriz Extracelular/metabolismo , Células Estrelladas Hepáticas/efectos de los fármacos , Células Estrelladas Hepáticas/metabolismo , Células Estrelladas Hepáticas/patología , Macrófagos del Hígado/efectos de los fármacos , Macrófagos del Hígado/metabolismo , Hepatopatías/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Miofibroblastos/patología , Fenotipo , Células Madre/efectos de los fármacos , Células Madre/metabolismoRESUMEN
SCOPE: Polyphenolic constituents of green tea (Camellia sinensis) have been shown to be potent scavengers of reactive oxygen species (ROS). Thus, this study was designed to assess its effects after liver ischemia-reperfusion. METHODS AND RESULTS: Fasted Sprague-Dawley rats were gavaged with different concentrations of green tea extract (GTE) 2 h before 90 min of warm ischemia of the left lateral liver lobe (30% of liver). Controls were given the same volume of Ringer's solution. A preparation of pentobarbital sodium (intraperitoneal) and ketamine (intramuscular) was used for anesthesia. After reperfusion, transaminases, liver histology, hepatic microcirculation, and both phagocytosis of latex bead particles as well as the expression of tumor necrosis alpha (TNF-α) to index cellular activation were investigated. Furthermore, the expression of superoxide dismutase (Mn-SOD) was assessed. After 90 min of warm ischemia aminotransferase (AST), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH) increased dramatically to 1946 ± 272/3244 ± 757 U/L, 1680 ± 134/2080 ± 379 U/L, and 7857 ± 1851/2036 ± 1193 U/L at 2/6 h, respectively. GTE (200 mg/kgbody weight) significantly prevented this increase in a dose-dependent manner by 21-51% at 2 h and 29-34% at 6 h, respectively. Histology confirmed the protective effects while both TNF-α expression and phagocytosis of latex beads by Kupffer cells (KCs) were significantly reduced. GTE intake significantly increased the expression of manganese superoxide dismutase. In vivo microscopy revealed improved acinar and sinusoidal perfusion after GTE. CONCLUSION: Preconditioning with a single oral dose of GTE ameliorates ischemia-reperfusion injury in liver. Decreased cellular activation and improved microcirculation are the proposed mechanisms.
Asunto(s)
Depuradores de Radicales Libres/administración & dosificación , Precondicionamiento Isquémico , Hígado/efectos de los fármacos , Extractos Vegetales/administración & dosificación , Daño por Reperfusión/prevención & control , Té/química , Isquemia Tibia/efectos adversos , Animales , Camellia sinensis/química , Relación Dosis-Respuesta a Droga , Femenino , Depuradores de Radicales Libres/uso terapéutico , Macrófagos del Hígado/efectos de los fármacos , Macrófagos del Hígado/patología , Hígado/irrigación sanguínea , Hígado/metabolismo , Hígado/patología , Microcirculación/efectos de los fármacos , Fagocitosis/efectos de los fármacos , Fitoterapia , Extractos Vegetales/uso terapéutico , Hojas de la Planta/química , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Flujo Sanguíneo Regional/efectos de los fármacos , Daño por Reperfusión/metabolismo , Daño por Reperfusión/patología , Superóxido Dismutasa/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Hepatotoxic side effects of neoadjuvant chemotherapy for colorectal liver metastases increase perioperative morbidity and mortality. Glycine protects liver from injury in various animal models. Thus, this study was designed to assess its effect on liver after chemotherapy. Sprague-Dawley rats (200-220 g) were fed a synthetic diet containing 5% glycine for 5 days. Subsequently, chemotherapy (FOLFIRI: irinotecan, folinic acid and fluorouracil, or FOLFOX: oxaliplatin, folinic acid and fluorouracil) was administered at standard doses. Transaminases, histology, immunohistochemistry and in vivo microscopy were used to index liver injury, to monitor intrahepatic microperfusion and activation of Kupffer cells. Glycine significantly decreased transaminases after chemotherapy to 25-50% of control values (p < 0.05). Microvesicular steatosis was significantly reduced from 18.5 ± 3.4 and 57.1 ± 8.6% in controls to 9.5 ± 1.8 and 37.7 ± 4.4% after FOLFIRI and FOLFOX, respectively. Furthermore, phagocytosis of latex beads was reduced by about 50%, while leukocyte adherence in central and midzonal subacinar zones decreased to 60-80% after glycine (p < 0.05). Glycine significantly reduced expression of inducible nitric oxide synthase after chemotherapy, while hepatic microcirculation was increased (p < 0.05). This study shows for the first time that glycine reduces chemotherapy-induced liver injury. The underlying mechanisms most likely include Kupffer cells and an improved intrahepatic microperfusion.
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Antineoplásicos/efectos adversos , Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , Suplementos Dietéticos , Hígado Graso/prevención & control , Glicina/administración & dosificación , Animales , Antineoplásicos/administración & dosificación , Camptotecina/administración & dosificación , Camptotecina/efectos adversos , Camptotecina/análogos & derivados , Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/mortalidad , Neoplasias Colorrectales/patología , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/prevención & control , Hígado Graso/etiología , Hígado Graso/metabolismo , Hígado Graso/patología , Femenino , Fluorouracilo/administración & dosificación , Fluorouracilo/efectos adversos , Inmunohistoquímica , Irinotecán , Macrófagos del Hígado/patología , Leucovorina/administración & dosificación , Leucovorina/efectos adversos , Hígado/efectos de los fármacos , Hígado/metabolismo , Hígado/patología , Microcirculación , Terapia Neoadyuvante/efectos adversos , Óxido Nítrico Sintasa de Tipo II/análisis , Compuestos Organoplatinos/administración & dosificación , Compuestos Organoplatinos/efectos adversos , Oxaliplatino , Fagocitosis , Ratas , Ratas Sprague-Dawley , Transaminasas/análisisRESUMEN
AIM OF STUDY: To investigate action mechanism of Yi Guan Jian Decoction on cirrhosis induced by CCl(4) in rats. MATERIAL AND METHODS: CCl(4) (3 mL/kg) for the first time and then olive oil CCl(4) solution 50% (2 mL/kg) was administered hypodermically to rats twice each week for 12 weeks. At the end of 8th week, rats were randomly divided into CCl(4) control group (n=10), Yi Guan Jian Decoction group (n=9) and Xiao Chai Hu Decoction group (n=9). Yi Guan Jian Decoction and Xiao Chai Hu Decoction were oral administrated per day respectively for 4 weeks, concomitantly continued CCl(4) administration. At 12th weekend, the rats were sacrificed for sampling and detection of liver function, histological changes of liver tissue, liver tissue hydroxyproline content and expression of alpha-SMA, CD68, MMP-13, TIMP-1, TIMP-2, Caspase-12, HGFalpha, MMP-2, MMP-9 and hepatocyte apoptotic index. RESULTS AND CONCLUSIONS: (1) Compared with that of normal rats, expression of alpha-SMA, CD68 and TIMP-1 in liver tissue of 8 week model group rats increases significantly (P<0.01), moreover further increased in the 12 week of model group. However, MMP-13, HGFalpha, TIMP-2 content decreases gradually and the statistical difference is seen between each time point (P<0.01). Activity of MMP-2, MMP-9, content of Caspase-12 and hepatocyte apoptotic index increased gradually at 4th, 8th, 12th week. (2) Compared to that of the same time point model group, activity of MMP-9 and contents of MMP-13, TIMP-2 and HGFalpha in Yi Guan Jian Decoction group improves significantly (P<0.01), and activity of MMP-2 and contents of alpha-SMA, TIMP-1, Caspase-12 and hepatocyte apoptotic index decreases significantly (P<0.01). This work suggests that Yi Guan Jian Decoction exerts significant therapeutic effect on CCl(4)-induced cirrhosis in rats, through mechanism of inhibiting hepatocytes apoptosis and hepatic stellate cells activation, and regulating the function of Kupffer cell. ETHNOPHARMACOLOGICAL RELEVANCE: This study investigates the mechanism of Yi Guan Jian against cirrhosis from aspect of heptocytes apoptosis and hepatic stellate cells activation. It suggest that although of unknown bioactive ingredients, mechanism of traditional Chinese medicine recipe against cirrhosis can be disclosed and of profound significance.
Asunto(s)
Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico , Cirrosis Hepática Experimental/tratamiento farmacológico , Hígado/efectos de los fármacos , Fitoterapia , Administración Oral , Animales , Apoptosis/efectos de los fármacos , Tetracloruro de Carbono , Células Estrelladas Hepáticas/efectos de los fármacos , Células Estrelladas Hepáticas/patología , Hepatocitos/efectos de los fármacos , Hepatocitos/patología , Hidroxiprolina/metabolismo , Macrófagos del Hígado/efectos de los fármacos , Macrófagos del Hígado/patología , Hígado/metabolismo , Hígado/patología , Cirrosis Hepática Experimental/patología , Masculino , Ratas , Ratas WistarRESUMEN
BACKGROUND: Propolis is a natural product collected by honey bees from various plant sources. We aimed to determine the possible effects of propolis on oxidative stress and hepatocyte apoptosis in experimental obstructive jaundice. METHODS: Thirty rats were divided into three groups: group I, sham-operated; group II, ligation and division of the common bile duct (BDL); group III, BDL followed by oral supplementation of propolis in a daily dose of 100 mg/kg. Liver samples were examined under the light microscope and transmission electron microscope. Hepatocyte apoptosis was quantitated using the transferase-mediated uridine nick end labeling (TUNEL) assay. Plasma and liver malondialdehyde (MDA) levels and glutathione peroxidase (GSH-Px) activities were measured. RESULTS: The plasma and liver levels of MDA were significantly lower in the propolis group than in the BDL group (p < 0.05 and 0.014, respectively). Although liver GSH-Px activities were significantly higher in the propolis group than in the BDL group (p < 0.001), there was no significant difference between the plasma GSH-Px activities of these groups (p > 0.05). In the propolis group, the enlargement of hepatocytes, dilatation of canaliculi and the edema regressed. The regenerating and normal hepatocytes were demonstrated. In the TUNEL assay, propolis administration reduced hepatocyte apoptosis. CONCLUSION: Propolis showed a significant hepatoprotective effect in this experimental obstructive jaundice model.
Asunto(s)
Antiinfecciosos/farmacología , Ictericia Obstructiva , Estrés Oxidativo/efectos de los fármacos , Própolis/farmacología , Animales , Apoptosis/efectos de los fármacos , Modelos Animales de Enfermedad , Glutatión Peroxidasa/metabolismo , Hepatocitos/efectos de los fármacos , Hepatocitos/patología , Hepatocitos/ultraestructura , Etiquetado Corte-Fin in Situ , Ictericia Obstructiva/tratamiento farmacológico , Ictericia Obstructiva/metabolismo , Ictericia Obstructiva/patología , Macrófagos del Hígado/efectos de los fármacos , Macrófagos del Hígado/patología , Macrófagos del Hígado/ultraestructura , Masculino , Malondialdehído/metabolismo , Microscopía Electrónica , Ratas , Ratas WistarRESUMEN
We tested the hypothesis that laminarin (LAM), a beta (1-3) polysaccharide extracted from brown algae, can modulate the response to a systemic inflammation. Male Wistar rats (n=7 per group) were fed a standard diet (control) or a diet supplemented with LAM for 25 days (5% during 4 days followed by 10% during 21 days). Thereafter, Escherichia coli lipopolysaccharides (LPS; 10 mg/kg i.p.) were injected and the animals were sacrificed 24 h after LPS challenge. The hypothermia, hyperglycemia and hypertriglyceridemia occurring early after LPS administration were less pronounced in LAM-treated rats than in controls. The increase in serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) activities - reflecting hepatic alterations - was lessened after LPS injection in LAM-treated rats compared to control rats. LAM treatment decreased serum monocytes number, nitrite (NO2) and tumor necrosis factor-alpha (TNF-alpha). LAM also modulated intra-hepatic immune cells: it lowered the occurrence of peroxidase-positive cells (corresponding to monocytes/neutrophils) and, in contrast, it increased the number of ED2-positive cells, corresponding to resident hepatic macrophages, i.e. Kupffer cells. In conclusion, the hepatoprotective effect of marine beta (1-3) glucan during endotoxic shock may be linked to its immunomodulatory properties. We propose that both lower recruitment of inflammatory cells inside the liver tissue and lower secretion of inflammatory mediators play a role in the tissue protective effect of LAM. These effects could be due to a direct effect of beta-glucan on immune cells, or to an indirect effect through their dietary fibre properties (fermentation in the gut).
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Suplementos Dietéticos , Endotoxemia/prevención & control , Factores Inmunológicos/farmacología , Lipopolisacáridos/farmacología , Hígado/efectos de los fármacos , Polisacáridos/farmacología , Alanina Transaminasa/sangre , Animales , Aspartato Aminotransferasas/sangre , Glucemia/efectos de los fármacos , Temperatura Corporal/efectos de los fármacos , Dinoprostona/sangre , Endotoxemia/inducido químicamente , Endotoxemia/patología , Glucanos , Factores Inmunológicos/uso terapéutico , Macrófagos del Hígado/efectos de los fármacos , Macrófagos del Hígado/patología , L-Lactato Deshidrogenasa/sangre , Recuento de Leucocitos , Hígado/metabolismo , Hígado/patología , Masculino , Dióxido de Nitrógeno/sangre , Tamaño de los Órganos/efectos de los fármacos , Peroxidasa/metabolismo , Polisacáridos/uso terapéutico , Ratas , Ratas Wistar , Bazo/efectos de los fármacos , Bazo/patología , Triglicéridos/sangre , Factor de Necrosis Tumoral alfa/sangreRESUMEN
OBJECTIVE: To observe the effects of Jianpi Huoxue Decoction, a compound Chinese herbal medicine, on Kupffer cell signal pathway activation in rats with liver injury induced by Lieber-Decarli liquid diet and lipopolysaccharide (LPS). METHODS: SD rats were divided into normal, control liquid diet, ethanol liquid diet and ethanol liquid diet plus Jianpi Huoxue Decoction group. Rats were administrated with Jianpi Huoxue Decoction or distilled water via gastrogavage for 4 weeks after administration with ethanol or control liquid diet for 2 weeks respectively. After that, rats in each group were stimulated with LPS via gastrogavage for 3.5 h and harvested. Alanine aminotransferase (ALT) in serum and triglyceride (TG) in liver were analyzed. Pathological changes in liver tissues were observed in HE staining section. Tumor necrosis factor-alpha(TNF-alpha) in portal vein plasma was assayed by ELISA. The protein expressions of CD68, Toll-like receptor 4 (TLR4), phosphorylation-I kappa B (P-I kappa B) and TNF-alpha in liver were evaluated with Western-blotting. RESULTS: After the treatment with Jianpi Huoxue Decoction, the pathologic changes in liver tissue were lightened, levels of ALT in serum, TG in liver and TNF-alpha in portal vein plasma were decreased, and the protein expressions of CD68, TLR4, P-IkappaB and TNF-alpha in liver were reduced. CONCLUSION: Jianpi Huoxue Decoction can inhibit Kupffer cell signal pathway activation in rats with liver injury induced by Lieber-Decarli liquid diet and LPS.
Asunto(s)
Medicamentos Herbarios Chinos/uso terapéutico , Macrófagos del Hígado/efectos de los fármacos , Hepatopatías Alcohólicas/tratamiento farmacológico , Fitoterapia , Alanina Transaminasa/sangre , Animales , Antígenos CD/metabolismo , Antígenos de Diferenciación Mielomonocítica/metabolismo , Western Blotting , Dieta , Medicamentos Herbarios Chinos/farmacología , Ensayo de Inmunoadsorción Enzimática , Macrófagos del Hígado/metabolismo , Macrófagos del Hígado/patología , Lipopolisacáridos , Hepatopatías Alcohólicas/etiología , Hepatopatías Alcohólicas/patología , Masculino , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 4/metabolismo , Triglicéridos/metabolismo , Factor de Necrosis Tumoral alfa/sangre , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Acute pancreatitis induces liver injury by upregulating Kupffer cell-derived Fas/FasL; on the other hand, acute pancreatitis induces apoptosis of Kupffer cells via NF-kappaB-dependent pathways. The balance between upregulation of Fas/FasL and Fas/FasL-induced apoptosis of its originator cell may determine the severity of pancreatitis-related liver injury. The aim of our study was to determine the role of p65 NF-kappaB/RelA in pancreatitis-induced Kupffer cell apoptosis. Acute pancreatitis was induced in NIH Swiss mice by a choline-deficient ethionine-supplement (CDE) diet. In vitro mouse Kupffer cell line was transfected with p65 siRNA and treated with pancreatic elastase to mimic pancreatitis. CDE pancreatitis upregulated nuclear translocation of p65 NF-kappaB/RelA, Fas/FasL, caspase-3, and DNA fragmentation in mice livers (all P < 0.001). In vitro, pancreatic elastase mimicked CDE-pancreatitis by upregulating nuclear translocation of p65 NF-kappaB/RelA, Fas/FasL, caspase-3, DNA fragmentation, and apoptosis in Kupffer cells (all P < 0.001). Transfection with p65 siRNA attenuated the elastase-induced nuclear translocation of p65 NF-kappaB/RelA, upregulation of Fas/FasL, caspase-3, DNA fragmentation, and apoptosis in Kupffer cells (all P < 0.001). Acute pancreatitis activates p65 NF-kappaB/RelA and induces apoptosis of Kupffer cells. Inhibition of p65NF-kappaB/RelA attenuates elastase-induced upregulation of proapoptotic pathways and apoptosis in Kupffer cells. The ability of Kupffer cells to autoregulate their stress response by inducing self-apoptosis warrants further investigation.
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Apoptosis/fisiología , Macrófagos del Hígado/fisiología , Pancreatitis/fisiopatología , Factor de Transcripción ReIA/fisiología , Enfermedad Aguda , Animales , Caspasa 3 , Caspasas/metabolismo , Fragmentación del ADN , Ensayo de Inmunoadsorción Enzimática , Proteína Ligando Fas , Femenino , Citometría de Flujo , Homeostasis/fisiología , Immunoblotting , Macrófagos del Hígado/patología , Glicoproteínas de Membrana/metabolismo , Ratones , Pancreatitis/patología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transfección , Translocación Genética , Factores de Necrosis Tumoral/metabolismo , Regulación hacia Arriba/fisiología , Receptor fas/metabolismoRESUMEN
Heat shock preconditioning (HPc) of fatty donor livers significantly increases recipient survival in rats. We investigated to what extent the blockade of Kupffer cells by gadolinium chloride (GdCl3) can mimic the effect of HPc and the involvement of liver CD4+ T lymphocytes in HPc. Fatty liver was experimentally induced in Lewis rats by a choline- and methionine-deficient diet. Fatty liver donors were pretreated with HPc (42.5 degrees C for 10 min), the Kupffer cell inhibitor GdCl3, or placebo (sham group). Donors were then harvested, stored in University of Wisconsin preservation solution for 12 h at 4 degrees C, and transplanted into normal syngeneic rats. Hepatic injury (alanine aminotransferase) and serum cytokines (interleukin-12p70, tumor necrosis factor-alpha, and interleukin-10) of recipients increased at 3 h, then decreased, and increased again at 24 h after transplantation. HPc treatment diminished both the early and later phases of this biphasic response and improved recipient survival. GdCl3 reduced these cytokines in the early but not the later phase and did not reduce neutrophil accumulation or improve the recipient survival. HPc, but not GdCl3 treatment, also reduced the number of liver CD4+ T lymphocytes and their interferon-gamma production. We conclude that HPc, but not GdCl3 treatment, prevents biphasic liver injury and the activation of liver CD4+ T lymphocytes in transplanted fatty donor livers.
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Linfocitos T CD4-Positivos/inmunología , Hígado Graso/cirugía , Supervivencia de Injerto/inmunología , Respuesta al Choque Térmico , Trasplante de Hígado , Acondicionamiento Pretrasplante/métodos , Enfermedad Aguda , Animales , Antiinflamatorios/farmacología , Linfocitos T CD4-Positivos/metabolismo , Citocinas/metabolismo , Modelos Animales de Enfermedad , Gadolinio/farmacología , Hipertermia Inducida , Macrófagos del Hígado/efectos de los fármacos , Macrófagos del Hígado/patología , Hígado/inmunología , Hígado/patología , Hígado/cirugía , Activación de Linfocitos , Masculino , Ratas , Ratas Endogámicas Lew , Daño por Reperfusión/inmunología , Daño por Reperfusión/prevención & controlRESUMEN
OBJECTIVE: To study the effects of Liangge San to the expression of CD14 and scaverger receptor(SR) in the kupffer cells of liver and the pathological changes of liver tissue of endotoxemia-mice. METHOD: The model was established with intravenous injection of lipopolysaccharide (LPS) and at the same time different dose Liangge San were given. The expression of CD14 and scaverger receptor were detected with immunohigtochemistry at the 2nd, 4th, 8th hour ofter injury and analyzed with computer image system, and the pathological changes of liver tissue were also observed. RESULT: At the three different hours, the expression of CD14 and scaverger receptor in macrophages of liver of LPS-injury group showed significant increase and significant decrease respectively, compared with that of the blank-control group (P < 0.01). The expression in dexamethasone group and Liangge San different dose groups were intermediate between those in injury group and those in control group. Compared with expression of LPS-injury group, those of dexamethasone group and Liangge San different dose groups showed significant differences (P < 0.01), especially that of Liangge San high dose group. Liver cells showed vacuole change. Changes of CD14 and SR expression were paralleled with the severity of liver damages of the mice. CONCLUSION: Liangge San can inhibite the up-regulation of CD14 expression and down-regulation of scaverger receptor expression in a dosage-dependent manner and also alleviate the damages of liver induced by LPS.
Asunto(s)
Medicamentos Herbarios Chinos/farmacología , Endotoxemia/metabolismo , Receptores de Lipopolisacáridos/metabolismo , Hígado/metabolismo , Receptores Depuradores/metabolismo , Animales , Antiinflamatorios/farmacología , Dexametasona/farmacología , Relación Dosis-Respuesta a Droga , Combinación de Medicamentos , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/aislamiento & purificación , Endotoxemia/inducido químicamente , Endotoxemia/patología , Femenino , Macrófagos del Hígado/metabolismo , Macrófagos del Hígado/patología , Lipopolisacáridos , Hígado/patología , Ratones , Plantas Medicinales/químicaRESUMEN
OBJETIVO: Avaliação morfológica do fígado e baço de ratos submetidos à oxigenoterapia hiperbárica após a ligadura das veias hepáticas. MÉTODO: Foram utilizados 30 animais machos adultos da espécie Holtzman, distribuídos aleatoriamente em dois grupos de 15 animais cada, assim designados: grupo 1 - ligadura das veias hepáticas; grupo 2 - ligadura das veias hepáticas associada à oxigenoterapia hiperbárica. Todos os animais foram submetidos à anestesia geral por meio de solução contendo cloridrato de cetamina (40 mg/ml) e cloridrato de meperidina (10 mg/ml) na dose de 50 mg/kg/peso, laparotomia mediana e ligadura das veias hepáticas. A oxigenoterapia hiperbárica foi aplicada nos animais do grupo 2, a partir da oitava hora do pós-operatório, por 120 minutos, sendo 90 minutos sob pressão de 2,5 atmosferas e 15 minutos no início e final da terapêutica, para promover a compressão e descompressão gradativa no período de 20 dias consecutivos. No 21° dia de pós-operatório, os animais foram mortos por inalação de éter e submetidos à laparotomia e extirpação dos fígados e baços para exame histológico. Foram comparados os resultados da histologia hepática e esplênica aplicando-se o teste exato de Fisher, considerando-se a diferença significante de P < 0,05. RESULTADOS: Os exames histológicos dos fígados e baços dos animais dos grupos 1 e 2 mostraram as seguintes alterações: presença de trombose nas veias hepática, porta e centro-lobular em cinco (33,3 por cento) animais do grupo 1 e ausência no grupo 2; presença de necrose dos hepatócitos caracterizada como acentuada em sete animais (46,7 por cento) e leve em oito (53,3 por cento) animais do grupo 1, enquanto que, em todos os animais do grupo 2, esta alteração foi caracterizada como leve; presença de células de Kupffer muito proeminentes e hipertrofiadas em 14 (93,3 por cento) animais do grupo 1 e pouco proeminentes e hipertrofiadas em todos os animais do grupo 2; congestão da polpa vermelha considerada acentuada em seis (40 por cento) e moderada em nove (60 por cento) animais do grupo 1 e em todos os animais do grupo 2; hemossiderose moderada ou acentuada em 14 (93,3 por cento) animais do grupo 1 e leve em todos os animais do grupo 2. As análises estatísticas realizadas entre os dois grupos mostraram diferenças significativas em todas a variáveis estudadas (P < 0,05)...
Asunto(s)
Animales , Masculino , Ratas , Bazo/anatomía & histología , Oxigenoterapia Hiperbárica , Hígado/anatomía & histología , Trombosis , Ratas Sprague-Dawley , Hepatocitos/patología , Hemosiderosis , Venas Hepáticas , Macrófagos del Hígado/patología , LigaduraRESUMEN
The literature evidencing the role of iron in promoting a range of neoplasms in humans and animals prompted us to search for a possible association between chemically induced hemosiderosis and hemangiosarcomas in the liver of mice in selected studies conducted by the National Toxicology Program (NTP). Its historical control database was examined for studies in which treatment-related liver hemangiosarcoma was noted; 130 consecutive NTP studies in B6C3F1 mice from Technical Report (TR)-340 to TR-493 were evaluated. Three compounds (2-butoxyethanol, p-nitroaniline, and para-chloroaniline) were associated with a relatively high incidence of Kupffer cell pigmentation consisting of hemosiderin in both sexes; only the male mice developed a relatively low incidence of treatment-related hemangiosarcoma. With a fourth compound (o-nitroanisole), a relatively low incidence (16/50, high-dose males) of chemical-related hemosiderosis was noted, with no associated increase of hemangiosarcoma. Two chemicals (pentachlorophenol and tetrafluoroethylene) increased the incidence of liver hemangiosarcoma in male and female mice, with no increase in Kupffer cell pigmentation. The overall association between liver hemangiosarcoma and Kupffer cell pigmentation was highly significant (p < 0.001). The cause for hemosiderosis in all cases was the erythrocytic hemolytic effect of the compounds. The reason for the sex-increased susceptibility for development of hemangiosarcoma is unknown but may be due to a hormone-related, reduced antioxidative defensive capacity through modulation of the activities of antioxidative enzymes.