Correction: Crespo et al. Protective Effect of Protocatechuic Acid on TNBS-Induced Colitis in Mice Is Associated with Modulation of the SphK/S1P Signaling Pathway. Nutrients 2017, 9, 288.

In the original publication [...].

Melatonin modulates mitophagy, innate immunity and circadian clocks in a model of viral-induced fulminant hepatic failure.

The haemorrhagic disease virus (RHDV) is a non-cultivable virus that promotes in rabbits an acute disease which accomplishes many characteristics of an animal model of fulminant hepatic failure (FHF). Beneficial effects of melatonin have been reported in RHDV-infected rabbits. This study investigated whether protection against viral-derived liver injury by melatonin is associated with modulation of mitophagy, innate immunity and clock signalling. Rabbits were experimentally infected with 2 × 104 haemagglutination units of a RHDV isolate and killed at 18, 24 and 30 hours after infection (hpi). Melatonin (20 mg/kg body weight ip) was administered at 0, 12 and 24 hpi. RHDV infection induced mitophagy, with the presence of a high number of mitophagosomes in hepatocytes and increased expression of mitophagy genes. Greater expression of main innate immune intermediaries and inflammasome components was also found in livers with RHDV-induced FHF. Both mitophagy and innate immunity activation was significantly hindered by melatonin. FHF induction also elicited an early dysregulation in clock signalling, and melatonin was able to prevent such circadian disruption. Our study discloses novel molecular routes contributing to RHDV-induced damage progression and supports the potential of melatonin as a promising therapeutic option in human FHF.

Author Correction: The wide utility of rabbits as models of human diseases.

This article was originally published under a CC BY-NC-SA License, but has now been made available under a CC BY 4.0 License.

Melatonin modulates dysregulated circadian clocks in mice with diethylnitrosamine-induced hepatocellular carcinoma.

Disruption of circadian rhythms, which are regulated by the circadian clock machinery, plays an important role in different long-term diseases including hepatocellular carcinoma (HCC). Melatonin has been reported to alleviate promotion and progression of HCC, but the potential contribution of circadian clock modulation is unknown. We investigated the effects of melatonin in mice which received diethylnitrosamine (DEN) (35 mg/kg body weight ip) once a week for 8 weeks. Melatonin was given at 5 or 10 mg kg-1 d-1 ip beginning 4 weeks after the onset of DEN administration and ending at the sacrifice time (10, 20, 30, or 40 weeks). Liver expression of Bmal1, Clock, Npas2, Rorα, and Sirt1 increased, whereas Cry1, Per1, Per2, Per3, CK1ε, Rev-erbα, and Rev-erbß decreased following DEN administration. Melatonin treatment prevented changes in the expression of clock genes, and this effect was accompanied by an upregulation of the MT1 receptor and reduced levels of the hypoxia-inducible factors Hif-1α and Hif-2α. An increased expression of p21, p53, and PARP1/2, a higher Bax/Bcl-2 ratio, and a lower expression of Cyclin D1, CDK6, HSP70, HSP90, and GRP78 proteins were also observed in melatonin-treated mice. Melatonin significantly potentiated the suppression of proliferation and cell cycle arrest induced by the synthetic REV-ERB agonist SR9009 in human Hep3B cells, and BMAL1 knocking down attenuated the pro-apoptotic and antiproliferative effect of melatonin. Results support a contribution of changes in the circadian clock components to the beneficial effects of melatonin in HCC and highlight the usefulness of strategies modulating the circadian machinery in hepatocarcinogenesis.

The wide utility of rabbits as models of human diseases.

Studies using the European rabbit Oryctolagus cuniculus contributed to elucidating numerous fundamental aspects of antibody structure and diversification mechanisms and continue to be valuable for the development and testing of therapeutic humanized polyclonal and monoclonal antibodies. Additionally, during the last two decades, the use of the European rabbit as an animal model has been increasingly extended to many human diseases. This review documents the continuing wide utility of the rabbit as a reliable disease model for development of therapeutics and vaccines and studies of the cellular and molecular mechanisms underlying many human diseases. Examples include syphilis, tuberculosis, HIV-AIDS, acute hepatic failure and diseases caused by noroviruses, ocular herpes, and papillomaviruses. The use of rabbits for vaccine development studies, which began with Louis Pasteur's rabies vaccine in 1881, continues today with targets that include the potentially blinding HSV-1 virus infection and HIV-AIDS. Additionally, two highly fatal viral diseases, rabbit hemorrhagic disease and myxomatosis, affect the European rabbit and provide unique models to understand co-evolution between a vertebrate host and viral pathogens.

Protective Effect of Protocatechuic Acid on TNBS-Induced Colitis in Mice Is Associated with Modulation of the SphK/S1P Signaling Pathway.

(1) Background: The present study aimed to investigate whether beneficial effects of protocatechuic acid (PCA) are associated with inhibition of the SphK/S1P axis and related signaling pathways in a 2,4,6-trinitrobenzenesulfonic acid (TNBS) model of inflammatory bowel disease; (2) Methods: Colitis was induced in male Balb/c mice by intracolonic administration of 2 mg of TNBS. PCA (30 or 60 mg/kg body wt) was given intraperitoneally daily for five days; (3) Results: Administration of PCA prevented the macroscopic and microscopic damage to the colonic mucosa, the decrease in body weight gain and the increase in myeloperoxidase activity induced by TNBS. PCA-treated mice exhibited a lower oxidized/reduced glutathione ratio, increased expression of antioxidant enzymes and Nrf2 and reduced expression of proinflammatory cytokines. Following TNBS treatment mRNA levels, protein concentration and immunohistochemical labelling for SphK1 increased significantly. S1P production and expression of S1P receptor 1 and S1P phosphatase 2 were significantly elevated. However, there was a decreased expression of S1P lyase. Furthermore, TNBS-treated mice exhibited increased phosphorylation of AKT and ERK, and a higher expression of pSTAT3 and the NF-κB p65 subunit. PCA administration significantly prevented those changes; (4) Conclusions: Data obtained suggest a contribution of the SphK/S1P system and related signaling pathways to the anti-inflammatory effect of PCA.

[THE TEN MOST CITED ARTICLES OF THE JOURNAL "NUTRICION HOSPITALARIA"]. / LOS DIEZ ARTÍCULOS MÁS CITADOS DE LA REVISTA "NUTRICIÓN HOSPITALARIA".

After 36 years of continued publication of the journal Nutrición Hospitalaria, a list with the ten most cited articles published in it is elaborated. The top ten most cited articles in the world literature and stratification according to language, English or Spanish, subject, or period of time published are also analyzed. Nutr Hosp is the most important Ibero latin American nutrition journal. Nutr Hosp published 369 items in 2014 gaining the fourth position among all the world's journals devoted to nutrition. Article publication in English, or simultaneously in Spanish and English and Open Access policy probably benefit the number of citations.

Después de 36 años de publicación ininterrumpida de la revista Nutrición Hospitalaria, hemos recopilado los diez artículos más citados. Se muestra también la relación de los diez artículos con más citas a escala mundial; asimismo se estratifican las citaciones según el idioma, inglés o español, la temática o los años analizados. Nutr. Hosp. es, a nivel mundial, la revista de nutrición ibero-latinoamericana mejor valorada. Por el volumen de artículos publicados, con 369 ítems citables en 2014, Nutr Hosp se sitúa en cuarto lugar de todas las revistas de nutrición. Permitir la publicación de artículos en inglés o hacerlo simultáneamente en castellano y en inglés, así como estar en régimen de "Open Access" sin restricción de ningún tipo en la difusión desde el momento de aparición de los artículos, son probablemente elementos favorecedores de las citas.

[SOME CONSIDERATIONS ABOUT THE INTRINSIC VALUE OF THE IMPACT FACTOR OF SCIENTIFIC JOURNALS]. / ALGUNAS CONSIDERACIONES SOBRE EL VALOR INTRÍNSECO DEL FACTOR DE IMPACTO DE LAS REVISTAS CIENTÍFICAS.

The reason of higher number of citations of some articles is discussed. Some considerations about the journals' impact factor, its merits and its pitfalls are also made. Scientific journals' impact factor, popularized by the Institute for Scientific Information, has become an objective parameter for authors' evaluation and also for institutions and other related circumstances. There is no reason for the impact factor's gap between some English journals and those written in other languages. English journals probably benefit of the "Mathew's effect", according to which eminent scientists are more rewarded by similar contributions than others less known. It is paradoxical that most of the major achievements of our age do not appear among the 100 most cited articles. There is no homogeneity among all the articles appearing in each scientific journal: half of the articles are cited ten times more than the other half. However, those articles cited 0 times are credited like the better ones. Each article should be evaluated by its own citations, which would be its impact factor; the authors should be evaluated by their H index.

Se analiza el porqué de las citaciones de los artículos. Se realizan también algunas consideraciones sobre el factor de impacto de las revistas, sus ventajas y sus posibles defectos. El factor de impacto de las revistas, desde su popularización por el Institute for Scientific Information, ha tomado una gran importancia como parámetro objetivo de evaluación de las revistas científicas y, por extensión, de todo lo que las rodea. No hay correlación con el desfase en factores de impacto de algunas revistas anglosajonas y el de las revistas escritas en otros idiomas. Probablemente se benefician de publicar en inglés y del llamado "efecto Mateo", según el cual los investigadores científicos eminentes cosechan aplausos mucho más nutridos que otros investigadores, menos conocidos, por contribuciones equivalentes. Es paradójico también que los grandes descubrimientos de nuestra época no figuren entre los 100 artículos más citados. No hay tampoco una correlación entre todos los artículos aparecidos en una publicación y su factor de impacto; la mitad de los artículos de una revista son citados diez veces más que la otra mitad. Los artículos citados 0 veces reciben el mérito de los mejores. Lo ortodoxo sería utilizar en cada artículo el número de citas que recibe, que sería su propio factor de impacto y, para los autores, el índice H.

Los diez artículos más citados de la revista 'Nutrición Hospitalaria' / The ten most cited articles of the journal 'Nutricion Hospitalaria'

Después de 36 años de publicación ininterrumpida de la revista Nutrición Hospitalaria, hemos recopilado los diez artículos más citados. Se muestra también la relación de los diez artículos con más citas a escala mundial; asimismo se estratifican las citaciones según el idioma, inglés o español, la temática o los años analizados. Nutr. Hosp. es, a nivel mundial, la revista de nutrición ibero-latinoamericana mejor valorada. Por el volumen de artículos publicados, con 369 ítems citables en 2014, Nutr Hosp se sitúa en cuarto lugar de todas las revistas de nutrición. Permitir la publicación de artículos en inglés o hacerlo simultáneamente en castellano y en inglés, así como estar en régimen de 'Open Access' sin restricción de ningún tipo en la difusión desde el momento de aparición de los artículos, son probablemente elementos favorecedores de las citas (AU)

After 36 years of continued publication of the journal Nutrición Hospitalaria, a list with the ten most cited articles published in it is elaborated. The top ten most cited articles in the world literature and stratification according to language, English or Spanish, subject, or period of time published are also analyzed. Nutr Hosp is the most important Ibero latin American nutrition journal. Nutr Hosp published 369 items in 2014 gaining the fourth position among all the world’s journals devoted to nutrition. Article publication in English, or simultaneously in Spanish and English and Open Access policy probably benefit the number of citations (AU)

Algunas consideraciones sobre el valor intrínseco del factor de impacto de las revistas científicas / Some considerations about the intrinsic value of the impact factor of scientific journals

Se analiza el porqué de las citaciones de los artículos. Se realizan también algunas consideraciones sobre el factor de impacto de las revistas, sus ventajas y sus posibles defectos. El factor de impacto de las revistas, desde su popularización por el Institute for Scientific Information, ha tomado una gran importancia como parámetro objetivo de evaluación de las revistas científicas y, por extensión, de todo lo que las rodea. No hay correlación con el desfase en factores de impacto de algunas revistas anglosajonas y el de las revistas escritas en otros idiomas. Probablemente se benefician de publicar en inglés y del llamado 'efecto Mateo', según el cual los investigadores científicos eminentes cosechan aplausos mucho más nutridos que otros investigadores, menos conocidos, por contribuciones equivalentes. Es paradójico también que los grandes descubrimientos de nuestra época no figuren entre los 100 artículos más citados. No hay tampoco una correlación entre todos los artículos aparecidos en una publicación y su factor de impacto; la mitad de los artículos de una revista son citados diez veces más que la otra mitad. Los artículos citados 0 veces reciben el mérito de los mejores. Lo ortodoxo sería utilizar en cada artículo el número de citas que recibe, que sería su propio factor de impacto y, para los autores, el índice H (AU)

The reason of higher number of citations of some articles is discussed. Some considerations about the journals’ impact factor, its merits and its pitfalls are also made. Scientific journals’ impact factor, popularized by the Institute for Scientific Information, has become an objective parameter for authors’ evaluation and also for institutions and other related circumstances. There is no reason for the impact factor’s gap between some English journals and those written in other languages. English journals probably benefit of the 'Mathew’s effect', according to which eminent scientists are more rewarded by similar contributions than others less known. It is paradoxical that most of the major achievements of our age do not appear among the 100 most cited articles. There is no homogeneity among all the articles appearing in each scientific journal: half of the articles are cited ten times more than the other half. However, those articles cited 0 times are credited like the better ones. Each article should be evaluated by its own citations, which would be its impact factor; the authors should be evaluated by their H index (AU)

Melatonin suppresses activation of hepatic stellate cells through RORα-mediated inhibition of 5-lipoxygenase.

Liver fibrosis is scar tissue resulting from an uncontrolled wound-healing process in response to chronic liver injury. Liver damage generates an inflammatory reaction that activates hepatic stellate cells (HSC) that transdifferentiate from quiescent cells that control retinol metabolism to proliferative and migratory myofibroblasts that produce excessive amounts of extracellular matrix proteins, in particular collagen 1a1 (COL1A1). Although liver fibrosis is reversible, no effective drug therapy is available to prevent or reverse HSC activation. Melatonin has potent hepatoprotective properties in a variety of acute and chronic liver injury models and suppresses liver fibrosis. However, it remains unclear whether melatonin acts indirectly or directly on HSC to prevent liver fibrosis. Here, we studied the effect of melatonin on culture-activated rat HSC. Melatonin dose-dependently suppressed the expression of HSC activation markers Col1a1 and alpha-smooth muscle actin (αSMA, Acta2), as well as HSC proliferation and loss of lipid droplets. The nuclear melatonin sensor retinoic acid receptor-related orphan receptor-alpha (RORα/Nr1f1) was expressed in quiescent and activated HSC, while the membranous melatonin receptors (Mtrn1a and Mtrn1b) were not. The synthetic RORα agonist SR1078 more potently suppressed Col1a1 and αSma expression, HSC proliferation, and lipid droplet loss, while the RORα antagonist SR1001 blocked the antifibrotic features of melatonin. Melatonin and SR1078 inhibited the expression of Alox5, encoding 5-lipoxygenase (5-LO). The pharmacological 5-LO inhibitor AA861 reduced Acta2 and Col1a1 expression in activated HSC. We conclude that melatonin directly suppresses HSC activation via RORα-mediated inhibition of Alox5 expression, which provides novel drug targets to treat liver fibrosis.

Effects of dietary supplementation with lemon verbena extracts on serum inflammatory markers of multiple sclerosis patients / Efecto de la suplementación dietética con extractos de hierbaluisa en los marcadores de inflamación en suero de personas con esclerosis múltiple

Introduction: Inflammation is one of the main contributory factors to the etiopathogenesis of multiple sclerosis (MS). Dietary interventions with Lipia citriadora (lemon verbena) extracts have been proved to be effective in the prevention of inflammatory diseases. Objectives: The aim of this study is to evaluate the effect of lemon verbena supplementation in pro- and anti- inflammatory serum biomarkers of patients with different clinical subtypes of multiple sclerosis. Methods: The effect of lemon verbena supplementation (10% w/w verbascoside) was evaluated in a randomized, double-blinded placebo-controlled study with 30 participants classified in relapsing-remitting (n=10), primary progressive (n=5) and secondary progressive (n=15) MS presentations. Serum cytokine and C reactive protein levels were assessed in intervention and control groups for each MS clinical subtype after 28 days of dietary supplementation. Results: Serum levels of C reactive protein and 8 cytokines/ inflammatory (IFN-γ, IL-12, IL-23, IL-6, TNF-α, TGF-β, IL-4 and IL-10) markers were studied. Secondary progressive MS- supplemented patients showed C reactive protein concentrations significantly lower compared to the placebo group (p<0.005). IFN-γ levels decreased for all MS-treated groups whereas IL-12 diminished levels were observed for relapsing-remitting type (p<0.05). Anti-inflammatory cytokine concentrations of IL-4 (difference 2.98 ± 2.99 pg/mL) and IL-10 (difference 1.78 ± 5.54 pg/mL) increased in secondary progressive MS patients (p<0.05). Conclusion: The variation of several pro- and anti-inflammatory markers after supplementation suggests that lemon verbena extracts may affect cytokine profiles in multiple sclerosis. Further investigation on dietary components with antioxidant and anti-inflammatory properties may contribute to understand MS pathogenesis and ameliorate MS symptoms (AU)

Introducción: La inflamación es uno de los principales factores que contribuyen en la etiopatogénesis de la esclerosis múltiple (EM). Se ha demostrado que las intervenciones en la dieta con extractos de Lipia citriadora (hierbaluisa) son efectivas en la prevención de las enfermedades inflamatorias. Objectivos: El objetivo de este estudio es evaluar el efecto de la suplementación con extractos de hierbaluisa en los biomarcadores de inflamación en suero de pacientes con diferentes subtipos clínicos de esclerosis múltiple. Métodos: El efecto de la suplementación con hierbaluisa (10 % p/p verbascósido) se evaluó mediante un estudio aleatorizado de doble ciego controlado con grupo placebo, constituido por 30 participantes clasificados según la forma de presentación de EM en: remitentes-recaídas (n=10), primaria progresiva (n=5) y secundaria progresiva (n=15). Los niveles de citoquinas y proteína C reactiva en suero se valoraron en los grupos intervención y control de cada uno de los subtipos clínicos de EM después de 28 días de suplementación en la dieta. Resultados: Se estudiaron los niveles en suero de proteína C reactiva y de 8 citoquinas como biomarcadores deinflamación (IFN-γ, IL-12, IL-23, IL-6, TNF-α, TGF-β, IL-4 e IL-10). Los pacientes del grupo de intervención con EM secundaria progresiva presentaron concentraciones de proteína C reactiva significativamente más bajos comparados con el grupo placebo (p<0.005). Los niveles de IFN-γ disminuyeron en todos los grupos tratados a la vez que se detectaron niveles inferiores de IL-12 en las formas secundaria progresiva y remitente-recaídas (p<0.05). Las concentraciones de las citoquinas anti-inflamatorias: IL-4 (diferencia 2,98 ± 2,99 pg/mL) y IL-10 (diferencia 1,78 ± 5,54 pg/mL) aumentaron en los pacientes con EM secundaria progresiva (p<0.05). Conclusión: La variación en la concentración de varias citoquinas pro- y anti-inflamatorias después de la suplementación con los extractos de hierbaluisa puede afectar al perfil de las citoquinas en la esclerosis múltiple. La investigación futura de los componentes de la dieta con propiedades anti-inflamatorias y antioxidantes puede contribuir a entender la patógenesis de la esclerosis múltiple así como a disminuir sus síntomas (AU)

Autophagic response in the Rabbit Hemorrhagic Disease, an animal model of virally-induced fulminant hepatic failure.

The Rabbit Hemorrhagic Disease Virus (RHDV) induces a severe disease that fulfils many requirements of an animal model of fulminant hepatic failure. However, a better knowledge of molecular mechanisms contributing to liver damage is required, and it is unknown whether the RHDV induces liver autophagy and how it relates to apoptosis. In this study, we attempted to explore which signalling pathways were involved in the autophagic response induced by the RHDV and to characterize their role in the context of RHDV pathogenesis. Rabbits were infected with 2 × 104 hemmaglutination units of a RHDV isolate. The autophagic response was measured as presence of autophagic vesicles, LC3 staining, conversion of LC3-I to autophagosome-associated LC3-II and changes in expression of beclin-1, UVRAG, Atg5, Atg12, Atg16L1 and p62/SQSTM1. RHDV-triggered autophagy reached a maximum at 24 hours post-infection (hpi) and declined at 30 and 36 hpi. Phosphorylation of mTOR also augmented in early periods of infection and there was an increase in the expression of the endoplasmic reticulum chaperones BiP/GRP78, CHOP and GRP94. Apoptosis, measured as caspase-3 activity and expression of PARP-1, increased significantly at 30 and 36 hpi in parallel to the maximal expression of the RHDV capsid protein VP60. These data indicate that RHDV infection initiates a rapid autophagic response, perhaps in an attempt to protect liver, which associates to ER stress development and is independent from downregulation of the major autophagy suppressor mTOR. As the infection continues and the autophagic response declines, cells begin to exhibit apoptosis.

Effects of dietary supplementation with lemon verbena extracts on serum inflammatory markers of multiple sclerosis patients.

INTRODUCTION: Inflammation is one of the main contributory factors to the etiopathogenesis of multiple sclerosis (MS). Dietary interventions with Lipia citriadora (lemon verbena) extracts have been proved to be effective in the prevention of inflammatory diseases. OBJECTIVES: The aim of this study is to evaluate the effect of lemon verbena supplementation in pro- and anti- inflammatory serum biomarkers of patients with different clinical subtypes of multiple sclerosis. METHODS: The effect of lemon verbena supplementation (10% w/w verbascoside) was evaluated in a randomized, double-blinded placebo-controlled study with 30 participants classified in relapsing-remitting (n=10), primary progressive (n=5) and secondary progressive (n=15) MS presentations. Serum cytokine and C reactive protein levels were assessed in intervention and control groups for each MS clinical subtype after 28 days of dietary supplementation. RESULTS: Serum levels of C reactive protein and 8 cytokines/ inflammatory (IFN-γ, IL-12, IL-23, IL-6, TNF-α, TGF-ß, IL-4 and IL-10) markers were studied. Secondary progressive MS- supplemented patients showed C reactive protein concentrations significantly lower compared to the placebo group (p.

Introducción: La inflamación es uno de los principales factores que contribuyen en la etiopatogénesis de la esclerosis múltiple (EM). Se ha demostrado que las intervenciones en la dieta con extractos de Lipia citriadora (hierbaluisa) son efectivas en la prevención de las enfermedades inflamatorias. Objectivos: El objetivo de este estudio es evaluar el efecto de la suplementación con extractos de hierbaluisa en los biomarcadores de inflamación en suero de pacientes con diferentes subtipos clínicos de esclerosis múltiple. Métodos: El efecto de la suplementación con hierbaluisa (10 % p/p verbascósido) se evaluó mediante un estudio aleatorizado de doble ciego controlado con grupo placebo, constituido por 30 participantes clasificados según la forma de presentación de EM en: remitentes-recaídas (n=10), primaria progresiva (n=5) y secundaria progresiva (n=15). Los niveles de citoquinas y proteína C reactiva en suero se valoraron en los grupos intervención y control de cada uno de los subtipos clínicos de EM después de 28 días de suplementación en la dieta. Resultados: Se estudiaron los niveles en suero de proteína C reactiva y de 8 citoquinas como biomarcadores de inflamación (IFN-, IL-12, IL-23, IL-6, TNF-, TGF-, IL-4 e IL-10). Los pacientes del grupo de intervención con EM secundaria progresiva presentaron concentraciones de proteína C reactiva significativamente más bajos comparados con el grupo placebo (p.

Cardiotrophin-1 promotes a high survival rate in rabbits with lethal fulminant hepatitis of viral origin.

Rabbit hemorrhagic disease virus (RHDV) causes lethal fulminant hepatitis closely resembling acute liver failure (ALF) in humans. In this study, we investigated whether cardiotrophin-1 (CT-1), a cytokine with hepatoprotective properties, could attenuate liver damage and prolong survival in virus-induced ALF. Twenty-four rabbits were infected with 2 × 10(4) hemagglutination units of RHDV. Twelve received five doses of CT-1 (100 µg/kg) starting at 12 h postinfection (hpi) (the first three doses every 6 h and then two additional doses at 48 and 72 hpi), while the rest received saline. The animals were analyzed for survival, serum biochemistry, and viral load. Another cohort (n = 22) was infected and treated similarly, but animals were sacrificed at 30 and 36 hpi to analyze liver histology, viral load, and the expression of factors implicated in liver damage and repair. All infected rabbits that received saline died by 60 hpi, while 67% of the CT-1-treated animals survived until the end of the study. Treated animals showed improved liver function and histology, while the viral loads were similar. In the livers of CT-1-treated rabbits we observed reduction of oxidative stress, diminished PARP1/2 and JNK activation, and decreased inflammatory reaction, as reflected by reduced expression of tumor necrosis factor alpha, interleukin-1ß, Toll-like receptor 4, VCAM-1, and MMP-9. In addition, CT-1-treated rabbits exhibited marked upregulation of TIMP-1 and increased expression of cytoprotective and proregenerative growth factors, including platelet-derived growth factor B, epidermal growth factor, platelet-derived growth factor receptor ß, and c-Met. In conclusion, in a lethal form of acute viral hepatitis, CT-1 increases animal survival by attenuating inflammation and activating cytoprotective mechanisms, thus representing a promising therapy for ALF of viral origin.

Role of quercetin in preventing thioacetamide-induced liver injury in rats.

In hepatic toxicity induced in rats by two injections of thioacetamide (TAA, 350 mg/kg with an interval of 8 hr), the action of quercetin was investigated. After 96 hr, TAA administration resulted in hepatic necrosis, significant increases in serum transaminase activity, and increases in hepatic lipoperoxidation. Thioacetamide-induced hepatotoxicity also showed changes in antioxidant enzymes in the liver of rats, with alterations in p-ERK 1/2 (phosphorylated extracellular-signal related kinase 1/2) as well as an imbalance between proapototic protein Bax and anti-apoptotic protein Bcl-2 expression. With administration of the flavonoid quercetin (50 mg/Kg i.p.) for four consecutive days following TAA, serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activity were close to normal values in rats. Histological findings suggested that quercetin had a preventive effect on TAA-induced hepatic necrosis. Quercetin treatment caused significant decreases in lipid peroxide levels in the TAA-treated rats, with some changes in antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). Quercetin also inhibited the change of the p-ERK1/2 by TAA and significantly prevented the increase in Bax/Bcl-2 ratio, thus preventing apoptosis. Findings indicate that quercetin may have a preventive effect on TAA-induced hepatotoxicity by modulating the oxidative stress parameters and apoptosis pathway.

S-nitroso-N-acetylcysteine attenuates liver fibrosis in cirrhotic rats.

This study was aimed to investigate the molecular mechanisms underlying prevention of hepatic fibrosis by S-nitroso-N-acetylcysteine (SNAC), a nitric oxide donor that inhibits lipid peroxidation. Secondary biliary cirrhosis was induced by 4 weeks of common bile duct ligation (CBDL). Both sham-operated and CBDL animals received SNAC (6.0 micromol/kg/day) starting 2 weeks after surgery. SNAC treatment reduced the increase in blood enzyme activities (alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase), induced by CBDL. Histological changes were attenuated and there was a significant decrease in the area of liver fibrosis and in the activation of stellate cells measured by alpha-smooth muscle actin (alpha-SMA) immunostaining. The increase in TBARS concentration and hydroperoxide-induced chemiluminescence were also reduced by SNAC treatment. SNAC down-regulated expression of collagen 1 alpha, alpha-SMA, tumor necrosis factor-alpha, tumor growth factor-beta, metalloproteinase-2, metalloproteinase inhibitor 1, platelet-derived growth factor (PDGF), and PDGF receptor in CBDL rats. These effects were accompanied by inhibited activation of extracellular signal-regulated kinases, Jun amino-terminal kinases, p38 and Akt. Antifibrotic effects were more efficient than those of the free thiol NAC administered at a dose of 60 mumol/kg. In conclusion, results obtained indicate that SNAC, beyond its antioxidant capacity, exerts antifibrotic effects in rats with secondary biliary cirrhosis by down-regulating increased expression of genes and modulating intracellular signaling pathways that contribute to the accumulation of matrix proteins. Thus, SNAC may be an interesting candidate for the treatment of human fibrosis and cirrhosis.

An overview of animal models for investigating the pathogenesis and therapeutic strategies in acute hepatic failure.

Acute hepatic failure (AHF) is a severe liver injury accompanied by hepatic encephalopathy which causes multiorgan failure with an extremely high mortality rate, even if intensive care is provided. Management of severe AHF continues to be one of the most challenging problems in clinical medicine. Liver transplantation has been shown to be the most effective therapy, but the procedure is limited by shortage of donor organs. Although a number of clinical trials testing different liver assist devices are under way, these systems alone have no significant effect on patient survival and are only regarded as a useful approach to bridge patients with AHF to liver transplantation. As a result, reproducible experimental animal models resembling the clinical conditions are still needed. The three main approaches used to create an animal model for AHF are: surgical procedures, toxic liver injury and infective procedures. Most common models are based on surgical techniques (total/partial hepatectomy, complete/transient devascularization) or the use of hepatotoxic drugs (acetaminophen, galactosamine, thioacetamide, and others), and very few satisfactory viral models are available. We have recently developed a viral model of AHF by means of the inoculation of rabbits with the virus of rabbit hemorrhagic disease. This model displays biochemical and histological characteristics, and clinical features that resemble those in human AHF. In the present article an overview is given of the most widely used animal models of AHF, and their main advantages and disadvantages are reviewed.

Oxidative stress induced by Cremophor EL is not accompanied by changes in NF-kappaB activation or iNOS expression.

The effects of polyoxyethylenglycerol triricinoleate 35 (Cremophor EL, CrEL) on markers of oxidative stress, nuclear factor kappa B (NF-kappaB) activation and inducible nitric oxide synthase (iNOS) expression were studied in the liver of male Wistar rats. Animals were randomly divided into three groups. Group Cr1 received, i.p., CrEL at 0.046ml/kg daily for 7 days, group Cr2 received CrEL at 0.33ml/kg and the controls were injected with CrEL vehicle (saline solution with 25% ethanol). Both alanine transaminase (ALT) and aspartate transaminase (AST) serum activities were significantly increased in the Cr2 group (+16% and +25%, respectively). AST activity was also higher in the Cr1 group when compared to control animals (+20%). The cytosolic concentration of thiobarbituric acid reactive substances (TBARS) increased in both groups of rats receiving CrEL (Cr1: +24%; Cr2: +33%). Reduced glutathione (GSH) concentration was not significantly modified at any of the CrEL doses, but both the hepatic concentration of oxidised glutathione (GSSG) (Cr1: +37%; Cr2: +84%) and the GSH/GSSG ratio (Cr1: -21%; Cr2: -45%) were significantly modified. CrEL induced no significant NF-kappaB activation, changes in p50 and p65 NF-kappaB subunits or induction of iNOS protein. Data obtained indicate that although high doses of CrEL cause oxidative stress, this is not enough to induce changes in NF-kappaB activation or iNOS expression.

Effects of FK506 and rapamycin on generation of reactive oxygen species, nitric oxide production and nuclear factor kappa B activation in rat hepatocytes.

We investigated the effect of two immunosuppressant drugs, FK506 and rapamycin, on reactive oxygen species (ROS) generation, nitric oxide (NO) production, inducible nitric oxide synthase (iNOS) expression and nuclear factor kappa B (NF-kappaB) activation in lipopolysaccharide (LPS)-activated rat hepatocytes. Primary culture of rat hepatocytes was treated with LPS in the presence and absence of FK506 or rapamycin. LPS increased the release of lactate dehydrogenase (LDH) and nitrite into the culture medium. Western blot and reverse transcription-polymerase chain reaction analyses demonstrated increased levels of iNOS protein and mRNA. Both immunosuppressant agents inhibited the induction of iNOS mRNA and protein stimulated by LPS. ROS generation, assessed by flow cytometry using dichlorodihydrofluorescein diacetate, was significantly decreased by FK506 and rapamycin. Moreover, electrophoretic mobility shift assay experiments indicated that both drugs blocked the LPS-induced activation of NF-kappaB. Inhibitor kappa B protein levels were decreased by LPS and this effect was partly blocked by FK506 or rapamycin. In summary, both immunosuppressant agents decreased the intracellular generation of ROS and inhibited NO production and iNOS expression at mRNA level in association to NF-kappaB activation. In addition to its capacity to reduce acute allograft rejection, this study highlights the anti-inflammatory properties of FK506 and rapamycin.