RESUMEN
BACKGROUND AND PURPOSE: GPR119 activation has been suggested to improve hyperglycemia, dyslipidemia and hepatic steatosis. But its therapeutic potential for metabolic dysfunction-associated steatohepatitis (MASH) are underexplored. Here, we investigated the effects of DA-1241, a novel GPR119 agonist, on MASH and explored its underlying mechanism of anti-inflammatory effects. EXPERIMENTAL APPROACH: The in vivo anti-MASH effect was assessed by examining the preventive effect in MS-MASH and Ob-MASH mice and the therapeutic effect in MASH with severe hyperglycemia and diet-induced obese (DIO)-MASH mice. Histological and biochemical changes in liver tissue were assessed. Both plasma and hepatic biomarkers related to inflammation and fibrosis were comprehensively analyzed. To understand its mode of action, changes in NFκB signaling were determined in HepG2 and THP-1 cells. KEY RESULTS: DA-1241 attenuated MASH progression and alleviated the MASH phenotypes in MASH mouse models with different etiologies, regardless of glucose-lowering activity. In DIO-MASH mice, DA-1241 significantly reduced biochemical parameters related to steatosis, inflammation and fibrosis in the liver with reduced plasma liver enzymes. When used in combination with a dipeptidyl peptidase 4 (DPP4) inhibitor, DA-1241 further improved the MASH phenotype by increasing endogenous glucagon-like peptide-1 effect. Notably, DA-1241 alone and in combination reduced liver inflammation and restored inflammation-related hepatic gene expression, leading to remission of systemic inflammation as assessed by plasma inflammatory cytokines and chemokines. We demonstrated that DA-1241 reduces macrophage differentiation through downregulation of NFκB signaling by activating GPR119. CONCLUSION: Our data suggest the therapeutic potential of DA-1241, alone and in combination with a DPP4 inhibitor, for MASH.
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Inhibidores de la Dipeptidil-Peptidasa IV , Hígado Graso , Animales , Ratones , Antivirales , Fibrosis , Inflamación/tratamiento farmacológico , FN-kappa B/efectos de los fármacosRESUMEN
AIM: The aim was to study whether oral glucosamine hydrochloride (GlcN.HCl) or mucopolysaccharide protein (MucoP) has a structure-modifying effect on an anterior cruciate ligament transection (ACLT) rabbit model of osteoarthritis (OA). METHODS: OA was surgically induced in the right knees of rabbits by transection of the ACLT. The left knees served as a sham-operated control. The animals were divided into four groups (n = 6 each): negative control (phosphate buffered saline, orally), positive control (oral celecoxib 10 mg/kg body weight/day), GlcN.HCl (oral 100 mg/kg/day) and MucoP (oral 100 mg/kg/day). Experimental animals were sacrificed after 8 weeks of treatment and the distal femur was removed for macroscopic examination, histological assessment, and terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) assay of the OA rabbits. RESULTS: On gross morphology, severe lesions were observed in articular cartilage in the negative control group. In the GlcN.HCl and MucoP treatment groups, fibrillations and cartilaginous lesions were significantly (P < 0.05) decreased compared to the negative control group. In particular, degenerative changes in cartilage and chondrocyte cellularity were significantly reduced (P < 0.05) in the positive control (celecoxib) group, GlcN.HCl treatment group and MucoP treatment group compared with the negative control group. TUNEL assay showed that apoptotic chondrocytes were significantly suppressed in the celecoxib group. Similar significant (P < 0.05) results were seen in the GlcN.HCl group and MucoP group but apoptosis of chondrocytes were high in the negative control group. CONCLUSION: These data suggest that the protective effects of GlcN.HCl and MucoP may play a useful role in the clinical treatment of OA.
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Cartílago Articular/efectos de los fármacos , Condrocitos/efectos de los fármacos , Glucosamina/administración & dosificación , Glicosaminoglicanos/administración & dosificación , Articulaciones/efectos de los fármacos , Osteoartritis/tratamiento farmacológico , Administración Oral , Animales , Ligamento Cruzado Anterior/cirugía , Apoptosis/efectos de los fármacos , Cartílago Articular/diagnóstico por imagen , Cartílago Articular/patología , Celecoxib/farmacología , Condrocitos/patología , Inhibidores de la Ciclooxigenasa 2/farmacología , Modelos Animales de Enfermedad , Miembro Posterior , Articulaciones/diagnóstico por imagen , Articulaciones/patología , Osteoartritis/patología , Conejos , Factores de TiempoRESUMEN
Smad3 is a key mediator of the transforming growth factor (TGF)-ß1 signaling pathway that plays central role in inflammation and fibrosis. In present study, we evaluated the effect of Smad3 deficiency in Smad3-/- mice with carbon tetrachloride (CCl4)-induced liver fibrosis. The animals were received CCl4 or olive oil three times a week for 4 weeks. Histopathological analyses were performed to evaluate the fibrosis development in the mice. Alteration of protein expression controlled by Smad3 was examined using a proteomic analysis. CCl4-induced liver fibrosis was rarely detected in Smad3-/- mice compared to Smad3+/+. Proteomic analysis revealed that proteins related to antioxidant activities such as senescence marker protein-30 (SMP30), selenium-binding proteins (SP56) and glutathione S-transferases (GSTs) were up-regulated in Smad3-/- mice. Western blot analysis confirmed that SMP30 protein expression was increased in Smad3-/- mice. And SMP30 levels were decreased in CCl4-treated Smad3+/+ and Smad3-/- mice. These results indicate that Smad3 deficiency influences the proteins level related to antioxidant activities during early liver fibrosis. Thus, we suggest that Smad3 deteriorate hepatic injury by inhibitor of antioxidant proteins as well as mediator of TGF-ß1 signaling.
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Proteínas de Unión al Calcio/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Proteína smad3/genética , Animales , Tetracloruro de Carbono/toxicidad , Electroforesis en Gel Bidimensional , Glutatión Transferasa/metabolismo , Cirrosis Hepática/inducido químicamente , Cirrosis Hepática/metabolismo , Cirrosis Hepática/patología , Masculino , Ratones , Ratones Noqueados , Proteómica , Receptores de Superficie Celular/metabolismo , Índice de Severidad de la Enfermedad , Transducción de Señal , Proteína smad3/deficiencia , Proteína smad3/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Factor de Crecimiento Transformador beta1/metabolismo , Regulación hacia ArribaRESUMEN
Following liver injuries, hepatic stellate cells (HSCs) express α-SMA. Mitogen activated protein kinase (MAPK) signaling pathways mediate α-SMA expression in distinct cell types. However, the regulation of α-SMA expression by MAPKs in HSCs has been rarely studied. We aimed to study the role of MAPKs in the activation of HSCs during liver fibrosis. Liver fibrosis of rats was induced by carbon tetrachloride. HSC-T6 cells, murine embryonic fibroblasts, JNK1(-/-) and JNK2(-/-) cells were used for in vitro studies. Immunohistochemistry and immunoblot analysis were used. We have found that the expression of JNK and α-SMA co-localized in HSCs during liver fibrosis, but ERK and p38 expressed in macrophages. The expression of α-SMA was up-regulated by JNK1 and JNK2 in non-stress condition. Under TGF-ß stimulation, however, the level α-SMA expression was increased by only JNK1, but not significantly changed by JNK2. We suggest that JNKs are responsible for α-SMA regulation, and especially JNK1 has a major role in up-regulation of α-SMA expression in HSCs under stress condition induced by TGF-ß during liver fibrosis.
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Actinas/biosíntesis , Células Estrelladas Hepáticas/metabolismo , Cirrosis Hepática/metabolismo , Proteína Quinasa 8 Activada por Mitógenos/metabolismo , Proteína Quinasa 9 Activada por Mitógenos/metabolismo , Animales , Western Blotting , Tetracloruro de Carbono/toxicidad , Inmunohistoquímica , Inmunoprecipitación , Cirrosis Hepática/inducido químicamente , Cirrosis Hepática/patología , Masculino , Ratas , Ratas Sprague-Dawley , Transducción de Señal/fisiología , Transfección , Regulación hacia ArribaRESUMEN
Recently, senescence marker protein-30 (SMP30) knockout (KO) mice have been reported to be susceptible to apoptosis, however, the role of SMP30 has not been characterized in the small intestine. The aim of the present study is to investigate the role of SMP30 in the process of spontaneous and γ-radiation-induced apoptosis in mouse small intestine. Eight-week-old male wild-type (WT) mice and SMP30 KO mice were examined after exposure to 0, 1, 3, 5, and 9 Gy of γ-radiation. Apoptosis in the crypts of the small intestine increased in the 0 to 5 Gy radiated SMP30 KO and WT mice. Radiation-induced apoptosis and the BAX/Bcl-2 ratio in the SMP30 KO mice were significantly increased in comparison to each identically treated group of WT mice (p < 0.05). The levels of spontaneous apoptosis in both WT and KO mice were similar (p > 0.05), indicating that increased apoptosis of crypt cells of SMP30 KO by irradiation can be associated with SMP30 depletion. These results suggested that SMP30 might be involved in overriding the apoptotic homeostatic mechanism in response to DNA damage.
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Apoptosis , Proteínas de Unión al Calcio/deficiencia , Intestinos/patología , Péptidos y Proteínas de Señalización Intracelular/deficiencia , Traumatismos por Radiación/patología , Animales , Apoptosis/efectos de la radiación , Proteínas de Unión al Calcio/metabolismo , Susceptibilidad a Enfermedades , Rayos gamma , Inmunohistoquímica , Etiquetado Corte-Fin in Situ , Mucosa Intestinal/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados , Traumatismos por Radiación/metabolismo , Proteína X Asociada a bcl-2/metabolismoRESUMEN
This study investigated the effects of acetyl-L-carnitine (ALC) in secondarily-induced cerebral chronic ischemia models using rats with permanent ligation of bilateral common carotid arteries (BCCL) and spontaneously hypertensive rats (SHR). Additionally, we used normal aged rats as a primary dementia model. Chronic ALC administration at 100 mg/kg (p.o.) for 4 weeks significantly attenuated neurodegenerative changes. In groups receiving 50 mg/kg or 100 mg/kg, ALC inhibited the active astrocyte increase in cerebral tissues of both BCCL and SHR models. In BCCL rats, ALC administration (50 mg/kg or 100 mg/kg, p.o.) resulted in significant promotion of glutathione levels in brain tissues. We also confirmed behavioral improvement after ALC treatment (100 mg/kg for 8 weeks, p.o.) on learning-memory function using aged rats (18 months old) in a passive avoidance task and preservation of CA1 pyramidal neurons was coincided on histopathological observation. In conclusion, chronic ALC administration may ameliorate cerebral ischemia progress after a cerebrovascular disorder as well as spontaneous ageing-related cerebral dysfunction via hippocampal protection.
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Acetilcarnitina/uso terapéutico , Envejecimiento/efectos de los fármacos , Reacción de Prevención/efectos de los fármacos , Isquemia Encefálica/prevención & control , Trastornos de la Memoria/prevención & control , Fármacos Neuroprotectores/uso terapéutico , Acetilcarnitina/farmacología , Envejecimiento/metabolismo , Envejecimiento/patología , Animales , Isquemia Encefálica/metabolismo , Isquemia Encefálica/patología , Masculino , Trastornos de la Memoria/metabolismo , Trastornos de la Memoria/patología , Fármacos Neuroprotectores/farmacología , Distribución Aleatoria , Ratas , Ratas Endogámicas SHRRESUMEN
Senescent cells have been observed in certain aged or damaged tissues. However, the information about the effects of aging on liver cells is limited. In the present study, we have examined age-related histological changes in the livers of senescence marker protein knockout (SMP30-/-) mice, which are considered as a murine aging model due to the more sensitive response to apoptotic reagents and due to their shorter life span. In livers of old SMP30-/- mice, numerous hepatic stellate cells (HSCs) were hypertrophic and contained abundant microvesicular lipid droplets in cytoplasm. We have found that the expression of peroxisome proliferators-activated receptor γ (PPARγ), which is a protein related to lipid metabolism and HSC quiescence, was increased in hypertrophic HSCs by aging and vitamin C (VC) deficiency, whereas these phenomena were dramatically reduced by antioxidant treatment. Therefore, these prominent phenotypic changes can be considered as aging markers in the livers of animals which are subjected to antioxidant property evaluation.
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Deficiencia de Ácido Ascórbico/metabolismo , Deficiencia de Ácido Ascórbico/patología , Células Estrelladas Hepáticas/metabolismo , Células Estrelladas Hepáticas/patología , PPAR gamma/biosíntesis , Animales , Western Blotting , Senescencia Celular , Femenino , Inmunohistoquímica , Ratones , Ratones Noqueados , Regulación hacia ArribaRESUMEN
INTRODUCTION: Preservation of the cavernous nerves (CNs) during radical prostatectomy is crucial for the patient's erectile function. Despite advances in operative technique, the majority of men report compromised erectile function postprostatectomy or complete loss of potency due to CN trauma even with nerve-sparing modifications. AIM: This study was designed to investigate whether repeated dosing of udenafil, a phosphodiesterase type 5 inhibitor, helps to improve erectile function after CN injury. METHODS: Using the CN crush injury model, 8-week-old male Sprague Dawley rats were divided into the following groups; sham-operated group, bilateral CN crush injury exposed to either no udenafil (vehicle) or udenafil (5, 20 mg/kg) daily for two different durations (4 and 8 weeks, p.o.). MAIN OUTCOME MEASURES: At both time points, CN electrical stimulation was used to assess erectile function by measuring the intracavernous pressure. The expressions of hypoxia-inducible factor 1-alpha (HIF-1α), transforming growth factor-beta (TGF-ß1), nerve growth factor (NGF), endothelin B receptor (ET(B) ), endothelial nitric oxide synthase (eNOS), neuronal nitric oxide synthase (nNOS), and sonic hedgehog homolog (SHH) in penile tissue were examined. Immunohistochemical antibody staining was performed for NGF, eNOS, nNOS, CD31, and alpha-smooth muscle actin (α-SMA). Additionally, terminal deoxynucleotidyl transferase-mediated nick-end labeling assay was performed to quantify apoptosis and the tissue slides were stained for Masson's trichrome to assess the smooth muscle/collagen ratio. RESULTS: Udenafil improved erectile function in a dose- and time-dependent manner with the maximum erectile function recovery achieved by 20 mg/kg udenafil at an 8-week time point. CN injury increased the expression of HIF-1α, TGF-ß1, NGF, and ET(B) , however, decreased the expression of eNOS, nNOS, and SHH. Udenafil significantly suppressed these alterations. The results from the histological analyses show that udenafil markedly reduces apoptosis induced by CN injury and augments the smooth muscle/collagen ratio. CONCLUSIONS: CN injury induces significantly impaired erectile function and altered gene/protein expression. Chronic administration of udenafil preserves erectile function and has a beneficial role against the pathophysiological consequences of CN injury.
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Disfunción Eréctil/tratamiento farmacológico , Inhibidores de Fosfodiesterasa 5/uso terapéutico , Pirimidinas/uso terapéutico , Sulfonamidas/uso terapéutico , Animales , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Disfunción Eréctil/etiología , Proteínas Hedgehog/análisis , Subunidad alfa del Factor 1 Inducible por Hipoxia/análisis , Masculino , Factor de Crecimiento Nervioso/análisis , Óxido Nítrico Sintasa de Tipo I/análisis , Óxido Nítrico Sintasa de Tipo III/análisis , Erección Peniana/efectos de los fármacos , Pene/química , Pene/efectos de los fármacos , Pene/inervación , Inhibidores de Fosfodiesterasa 5/administración & dosificación , Pirimidinas/administración & dosificación , Ratas , Ratas Sprague-Dawley , Receptor de Endotelina B/análisis , Sulfonamidas/administración & dosificación , Factor de Crecimiento Transformador beta1/análisisRESUMEN
ENA Actimineral Resource A (ENA-A) is alkaline water that is composed of refined edible cuttlefish bone and two different species of seaweed, Phymatolithon calcareum and Lithothamnion corallioides. In the present study, ENA-A was investigated as an antioxidant to protect against CCl(4)-induced oxidative stress and hepatotoxicity in rats. Liver injury was induced by either subacute or chronic CCl(4) administration, and the rats had free access to tap water mixed with 0% (control group) or 10% (v/v) ENA-A for 5 or 8 weeks. The results of histological examination and measurement of antioxidant activity showed that the reactive oxygen species production, lipid peroxidation, induction of CYP2E1 were decreased and the antioxidant activity, including glutathione and catalase production, was increased in the ENA-A groups as compared with the control group. On 2-DE gel analysis of the proteomes, 13 differentially expressed proteins were obtained in the ENA-A groups as compared with the control group. Antioxidant proteins, including glutathione S-transferase, kelch-like ECH-associated protein 1, and peroxiredoxin 1, were increased with hepatocyte nuclear factor 3-beta and serum albumin precursor, and kininogen precursor decreased more in the ENA-A groups than compared to the control group. In conclusion, our results suggest that ENA-A does indeed have some protective capabilities against CCl(4)-induced liver injury through its antioxidant function.
Asunto(s)
Antioxidantes/farmacología , Tetracloruro de Carbono/toxicidad , Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , Minerales/farmacología , Preparaciones de Plantas/farmacología , Animales , Catalasa/metabolismo , Citocromo P-450 CYP2E1/metabolismo , Electroforesis en Gel Bidimensional , Inducción Enzimática/efectos de los fármacos , Glutatión/metabolismo , Glutatión Transferasa/metabolismo , Inmunohistoquímica , Péptidos y Proteínas de Señalización Intracelular , Proteína 1 Asociada A ECH Tipo Kelch , Peroxidación de Lípido/efectos de los fármacos , Espectrometría de Masas , Estrés Oxidativo/efectos de los fármacos , Peroxirredoxinas/metabolismo , Proteínas/metabolismo , Ratas , Especies Reactivas de Oxígeno/metabolismoRESUMEN
BACKGROUND AND AIMS: The binucleation of hepatocytes, which was known as an important feature of liver growth and physiology, has been reported to be increased during the chronic oxidative injury stage and has been regarded as an age-related change of hepatic structures. Therefore, we investigated the binuclearity pattern in the livers of senescence marker proteins-30 (SMP30) knock-out (KO) mice compared with wild-type (WT) mice and vitamin C-treated KO (KO + VC) mice. METHODS: The WT, KO and KO + VC mice were fed a vitamin C free diet and VC(+) group mice were given vitamin C water containing 1.5 g/L of vitamin C, whereas VC(-) group was given normal drinking water without vitamin C, for 16 weeks. RESULTS: In microscopic examination, the livers of KO mice showed a significantly increased number of binuclear hepatocytes compared with that of WT mice and KO + VC mice. KO mice also showed the most increased expression level of CYP2E1 and PCNA determined by immunohistochemistry and immunoblot analysis. Moreover, KO mice indicated the highest level of serum alanine aminotransferase and aspartate aminotransferase level in serum biochemical analysis. Accordingly, significantly decreased levels of reactive oxygen species, MDA (malondialdehyde) and HAE (4-hydroxyalkenals) were detected in KO + VC mice compared with KO mice. CONCLUSION: Therefore, it is concluded that vitamin C deficiency induces an increase of CYP2E1 expression and elevated ROS production, which causes oxidative liver injury and the elevation of hepatocyte binucleation in SMP30 KO mice.
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Deficiencia de Ácido Ascórbico/metabolismo , Núcleo Celular/patología , Citocromo P-450 CYP2E1/metabolismo , Hepatocitos/metabolismo , Hepatocitos/patología , Análisis de Varianza , Animales , Peso Corporal , Proteínas de Unión al Calcio/genética , Hepatocitos/ultraestructura , Péptidos y Proteínas de Señalización Intracelular/genética , Peroxidación de Lípido/efectos de los fármacos , Masculino , Ratones , Ratones Noqueados , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Regulación hacia ArribaRESUMEN
Our earlier report has shown that Helicobacter pylori promoted hepatic fibrosis in a murine model. Herein, in order to elucidate the mechanism by which H. pylori accelerate liver fibrosis, the authors investigated the changes in expression levels of mitogen-activated protein kinases (MAPKs), p53-related proteins, antioxidants, and proinflammatory cytokines in liver samples. H. pylori infection enhanced CCl4-induced MAP kinase activation and p53 signaling pathway as well as Bax- and proliferating-cell nuclear antigen expressions, whereas H. pylori alone induced neither of these expressions nor hepatic fibrosis. Moreover, mRNA expressions of inflammatory cytokines, glutathione peroxidase expression, and the proliferative index were strongly augmented in livers of the H. pylori with CCl4 treatment group compared with those of the CCl4-alone treatment group, whereas there was no difference in apoptotic index between the two groups. Interestingly, H. pylori treatment increased the number of α-fetoprotein-expressing hepatocytes independently of CCl4 intoxication. In vitro analyses, using an immortalized rat hepatic stellate cell (HSC) line, revealed that H. pylori lysates increased the proliferation of HSCs, which was boosted by the addition of transforming growth factor-beta1 (TGF-ß1). Furthermore, the treatment of H. pylori lysates promoted the translocation of nuclear factor kappa-light-chain enhancer of activated B cells (NF-κB) into the nucleus based on an increase in the degradation of NF-κB inhibitor alpha, in the presence of TGF-ß1, as did H2O2 treatment. In conclusion, H. pylori infection along with an elevated TGF-ß1 may accelerate hepatic fibrosis through increased TGF-ß1-induced pro-inflammatory signaling pathways in HSCs. Moreover, H. pylori infection might increase the risk of TGF-ß1-mediated tumorigenesis by disturbing the balance between apoptosis and proliferation of hepatocytes.
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Infecciones por Helicobacter/patología , Helicobacter pylori , Cirrosis Hepática/microbiología , Hígado/patología , Transducción de Señal , Animales , Apoptosis , Línea Celular , Proliferación Celular , Femenino , Infecciones por Helicobacter/metabolismo , Proteínas I-kappa B/metabolismo , Inflamación/microbiología , Hígado/microbiología , Cirrosis Hepática/metabolismo , Cirrosis Hepática/patología , Ratones , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Inhibidor NF-kappaB alfa , FN-kappa B/metabolismo , Ratas , Factor de Crecimiento Transformador beta1/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , alfa-Fetoproteínas/metabolismoRESUMEN
The present study reports a case of a 5-month-old female adrenomedullin (AM) heterozygous (+/-) mouse that presented a mass of leiomyosarcoma found in the right shoulder girdle region. The neoplastic mass extended to the sternal region and showed hemorrhages, congestion and necrotic foci. The excised tumor with a diameter of 2.5cm was firm, ill-demarcated and had focally infiltrated the surrounding muscles. The cut surface was homogeneously whitish with multi-focal reddish lesions. Microscopically, the tumor composed of variable fascicles of spindle-shaped cells with pleomorphic and cigar-shaped nuclei. The nuclei were round and elongated. Metastasis of tumor cell to skeletal muscle was frequently observed. Immunohistochemically, desmin, vimentin and alpha-smooth muscle actin (alpha-SMA) were demonstrated in neoplastic cells but tumor cells were negative for cytokeratin (CK) and S-100. Based on gross finding, microscopical examination and immunohistochemistry, the present case was diagnosed as a subcutaneous leiomyosarcoma.
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Adrenomedulina/genética , Leiomiosarcoma/veterinaria , Neoplasias de los Tejidos Blandos/veterinaria , Animales , Femenino , Heterocigoto , Inmunohistoquímica , Leiomiosarcoma/genética , Leiomiosarcoma/patología , Ratones , Ratones Endogámicos C57BL , Neoplasias de los Tejidos Blandos/genética , Neoplasias de los Tejidos Blandos/patología , Tejido Subcutáneo/patologíaRESUMEN
Helicobacter pylori infection has been reported to be very common in patients with chronic liver diseases, including cirrhosis. To elucidate the pathological effect of H. pylori infection on the progression of hepatic fibrosis, C57BL/6 mice and Sprague-Dawley rats were orally inoculated with H. pylori, and hepatic fibrosis was induced with carbon tetrachloride (CCl(4)) administration. We observed the histopathological changes and the presence of H. pylori genes by PCR in the liver. Significant increase in the fibrotic score as well as in serum alanine aminotransferase and aspartate aminotransferase levels was shown in the CCl(4)+H. pylori group compared with that in the CCl(4)-treated group. Compared with the CCl(4)-treated group, alpha-smooth muscle actin and transforming growth factor-beta1 were enhanced; however, senescence marker protein-30, a multifunctional protein protecting hepatocytes against oxidative stress and apoptosis, was suppressed in the CCl(4)+H. pylori group. The 16S rRNA (400 bp) was demonstrated by PCR for H. pylori genes from genomic DNA extracted from the liver, and H. pylori-infected mice showed 93.8% (15 of 16) seropositivity by contrast with seronegativity in all H. pylori-noninfected mice. In addition, immunohistochemical study against H. pylori showed positive antigen fragments in the liver of the infected groups. Consequently, our data suggest that H. pylori infection could be an important contributing infectious factor to the development of liver cirrhosis.
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Infecciones por Helicobacter/complicaciones , Helicobacter pylori/patogenicidad , Cirrosis Hepática Experimental/microbiología , Alanina Transaminasa/sangre , Animales , Aspartato Aminotransferasas/sangre , Biomarcadores/metabolismo , Proteínas de Unión al Calcio/genética , Proteínas de Unión al Calcio/metabolismo , Tetracloruro de Carbono/toxicidad , Progresión de la Enfermedad , Regulación Bacteriana de la Expresión Génica , Infecciones por Helicobacter/patología , Helicobacter pylori/genética , Péptidos y Proteínas de Señalización Intracelular/genética , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Cirrosis Hepática Experimental/sangre , Cirrosis Hepática Experimental/patología , Ratones , Ratones Endogámicos C57BL , ARN Ribosómico 16S/genética , Ratas , Ratas Sprague-DawleyRESUMEN
A 1.5-year-old female Persian cat was presented for inappetence and azotemia. Ultrasonography and urography revealed multiple abnormalities involving the genitourinary tract, including agenesis of the right kidney and ureter. Gross examination of the abnormal uterus revealed segmental aplasia of right caudal uterine horn causing cranial distension with fluid, a normal left uterine horn, and both normal ovaries. Microscopically, endometrial glands of the right uterine horn were markedly decreased in number. The right uterine horn was hemorrhagic suggesting estrus. This is the first report of this combination of urinary and uterus abnormalities in the veterinary literature.
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Enfermedades de los Gatos/congénito , Riñón/anomalías , Uréter/anomalías , Anomalías Urogenitales/veterinaria , Útero/anomalías , Amoxicilina/uso terapéutico , Animales , Antibacterianos/uso terapéutico , Enfermedades de los Gatos/diagnóstico , Enfermedades de los Gatos/terapia , Gatos , Femenino , Riñón/diagnóstico por imagen , Ultrasonografía , Uréter/diagnóstico por imagen , Anomalías Urogenitales/diagnóstico , Anomalías Urogenitales/terapia , Urografía/veterinaria , Útero/patologíaRESUMEN
Apoptosis occurs early after irradiation and may be a good indicator of radiation damages. Since elevated levels of TGF-beta are associated with radiation-induced inflammation, the null mice of Smad3, a key downstream mediator of TGF-beta, show accelerated healing of irradiated injury. In order to evaluate resistance to radiation-induced liver injuries in Smad3-null mice, we determined the occurrence of apoptosis and the expression of senescence marker protein-30 (SMP30), as an anti-apoptotic marker, after irradiation to the liver. The livers of Smad3-mutant mice were exposed to local irradiation of 15 gray, from a (60)Co-gamma radiation. One week after irradiation, in Smad3-KO mice, radiation-induced apoptosis was at lower levels compared to those of irradiated WT mice. These findings were well matched with the expression of CYP2E1, which plays a role in hepatic injuries produced by oxidative stress. In addition, antioxidant related protein, the SMP30 levels were reduced by gamma irradiation in both groups. Interestingly, the increased expression of SMP30 expression in Smad3-KO mice liver was preserved at a higher level than that of the WT mice after irradiation. Therefore, these results suggest that the interruption of TGF-beta signaling by deletion of Smad3 brings about inhibition of hepatic apoptosis after ionizing irradiation. Moreover, the protective effect to ionizing radiation might be in correlation with the overexpression of SMP30 in the Smad3-null mice, which may act as an anti-apoptotic signaling molecule. The alteration of SMP30 by interruption of Smad3 might be a useful therapeutic target and diagnostic marker for radiation-induced liver damages.
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Apoptosis/fisiología , Apoptosis/efectos de la radiación , Proteínas de Unión al Calcio/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Hígado/metabolismo , Hígado/efectos de la radiación , Proteína smad3/metabolismo , Animales , Hígado/citología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Dosis de Radiación , Tolerancia a Radiación/fisiología , Proteína smad3/genética , Regulación hacia Arriba/efectos de la radiaciónRESUMEN
We report a case of primary biliary cirrhosis (PBC) that occurred in a 24-month-old male C57BL/6 mouse infected with Helicobacter pylori (H. pylori). Microscopically, the portal tract in the liver showed nonsuppurative destructive cholangitis with variable cytologic distortion of the epithelial cells and peribiliary lymphoplasmacytic infiltration. Immunohistochemistry using alpha-smooth muscle actin demonstrated fibrous bands associating with the wall of vasculature. The level of serum antivacuolating toxin IgG in this mouse showed the highest value (optical density=2.1470) of the H. pylori-infected group (n=13) (optical density=1.7168+/-0.1759, mean+/-SD). Spontaneously developed PBC-like lesions in C57BL/6 mice have been reported by several authors. However, this case strikingly resembles human PBC with its characterized histological features. Therefore, we propose that the increase in vacuolating toxin caused by H. pylori infection may be related to the development of PBC by molecular mimicry.
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Infecciones por Helicobacter/complicaciones , Helicobacter pylori , Cirrosis Hepática Biliar/microbiología , Actinas/análisis , Animales , Proteínas Bacterianas/sangre , Infecciones por Helicobacter/sangre , Infecciones por Helicobacter/patología , Inmunohistoquímica , Hígado/patología , Cirrosis Hepática Biliar/sangre , Cirrosis Hepática Biliar/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Organismos Libres de Patógenos EspecíficosRESUMEN
A 3-year-old, female Spitz, was presented due to lack of response to therapies with a 6-month history of skin lesions characterized by diffuse erythema and scaling on the dorsal trunk. Physical examination revealed the dog was active and healthy. Skin culture isolated no fungus. Histological examination of skin biopsy specimens revealed interface dermatitis with hydropic degeneration of the basal layers, predominant plasmacytic perivascular accumulation in the dermis, and intensive plasma cell-rich interface mural folliculitis. Moderate CD3-positive lymphocytes infiltrated the superficial dermis. This report may provide unique information of canine discoid lupus erythematosus in an unusual breed with atypical cutaneous lesions.
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Enfermedades de los Perros/patología , Lupus Eritematoso Discoide/veterinaria , Animales , Perros , Femenino , Inmunohistoquímica , Lupus Eritematoso Discoide/patología , Especificidad de la EspecieRESUMEN
Arazyme is a novel protease produced by the HY-3 strain of Aranicola proteolyticus, which is a Gram-negative aerobic bacterium that has been isolated from the intestine of the spider Nephila clavata. This study focused on the hepatoprotective effect of Arazyme on carbon tetrachloride (CCl4)-induced acute hepatic injury in senescence marker protein 30 (SMP30) knock-out (KO) mice and SMP30 wild-type (WT) mice. WT mice and SMP30 KO mice were divided into eight groups as follows: (i) two negative control groups (G1, G5) which were treated with a single intraperitoneal (i.p.) olive oil injection. (ii) Two positive control groups (G2, G6) which received a single i.p. CCl4 (0.4mL/kg) injection. (iii) Two vitamin C-treated groups (G3, G7) which received a single oral administration of vitamin C (100mg/kg) and were injected with a single i.p. CCl4 (0.4mL/kg). (iv) Two Arazyme-treated groups (G4, G8) which received a single oral administration of Arazyme (500mg/kg) and were injected with a single i.p. CCl4 (0.4mL/kg). Through present study, we could find that Arazyme-treated groups showed decreased degree of liver injury, increased expression of SMP30, decreased expression of phospho-Smad3 (p-Smad3), elevated expression of antioxidant proteins including sorbitol dehydrogenase, dihydropteridine reductase (DHPR), dehydrofolate reductase (DHFR), NADH dehydrogenase, glutathione S-transferase kappa 1 (GSTK1) and phospholipid hydroperoxide glutathione peroxidase (PHGPx) compared with non-Arazyme-treated groups. Therefore, it is concluded that Arazyme plays a significant role in protecting injured hepatocytes by increasing the expression of SMP30, inhibiting the transforming growth factor-beta (TGF-beta)/Smad pathway and elevating the expression of antioxidant proteins.
Asunto(s)
Proteínas de Unión al Calcio/genética , Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , Péptidos y Proteínas de Señalización Intracelular/genética , Hígado/efectos de los fármacos , Metaloproteasas/farmacología , Animales , Antioxidantes/farmacología , Ácido Ascórbico/farmacología , Proteínas de Unión al Calcio/metabolismo , Intoxicación por Tetracloruro de Carbono , Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Quimioprevención , Modelos Animales de Enfermedad , Femenino , Técnica del Anticuerpo Fluorescente Indirecta , Técnicas para Inmunoenzimas , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Hígado/patología , Masculino , Ratones , Ratones Noqueados , Oxidorreductasas/metabolismo , Proteómica , Serratia/enzimología , Proteína smad3/metabolismo , Organismos Libres de Patógenos Específicos , Factor de Crecimiento Transformador beta/metabolismo , Regulación hacia ArribaRESUMEN
Helicobacter pylori vacuolating cytotoxin A (VacA) has been considered as an apoptosis-inducing factor. Here, we investigated the mechanism of VacA-induced apoptosis in relation to the defense mechanism and MAP kinases pathway in gastric epithelial cells. AGS cells exposed to enriched VacA extracts affected the level of SOD-1 and villin. We further investigated the role of VacA in those inductions using a functional recombinant VacA (rVacA). Activation of p38 MAPK and Bax dimerization by rVacA were increased in a dose-dependent manner. rVacA-induced ERK1/2 MAPK activation was maximal at 30 min and 4 h and 1-4 microg/ml of rVacA. rVacA-induced SOD-1 expression was considerably diminished by inhibiting ERK1/2 MAPK and it was slightly increased by inhibiting p38 MAPK. rVacA increased or decreased villin expression depending on dose and exposure time and its expression was mainly appeared in the contractile actin ring of the dividing cells. Despite its cytoprotective effect, SB-203580, a p38 inhibitor, was unlikely to reduce VacA-induced Bax dimerization and rather inhibited villin and Bcl2 expression, indicating that p38 may also play a role in cell proliferation or differentiation for survival after VacA intoxication. Furthermore, p38 inhibitor accelerated rVacA-induced cell death after exposure of AGS cells to H(2)O(2) but ERK1/2 inhibitor protected cells from H(2)O(2) insult. These results suggest that SOD-1 and villin are expressed differentially upon VacA insult depending on dose and exposure time via ERK and p38 MAP kinases; decrease in SOD-1 and villin expression coupled with Bax dimerization leads to apoptosis of gastric epithelial cells.