transforming-growth-factor-beta and Liver-Diseases--Alcoholic

Chronic intake of alcohol undoubtedly overwhelms the structural and functional capacity of the liver by initiating complex pathological events characterized by steatosis, steatohepatitis, hepatic fibrosis and cirrhosis. Subsequently, these initial pathological events are sustained and ushered into a more complex and progressive liver disease, increasing the risk of fibro-hepatocarcinogenesis. These coordinated pathological events mainly result from buildup of toxic metabolic derivatives of alcohol including but not limited to acetaldehyde (AA), malondialdehyde (MDA), CYP2E1-generated reactive oxygen species, alcohol-induced gut-derived lipopolysaccharide, AA/MDA protein and DNA adducts. The metabolic derivatives of alcohol together with other comorbidity factors, including hepatitis B and C viral infections, dysregulated iron metabolism, abuse of antibiotics, schistosomiasis, toxic drug metabolites, autoimmune disease and other non-specific factors, have been shown to underlie liver diseases. In view of the multiple etiology of liver diseases, attempts to delineate the mechanism by which each etiological factor causes liver disease has always proved cumbersome if not impossible. In the case of alcoholic liver disease (ALD), it is even more cumbersome and complicated as a result of the many toxic metabolic derivatives of alcohol with their varying liver-specific toxicities. In spite of all these hurdles, researchers and experts in hepatology have strived to expand knowledge and scientific discourse, particularly on ALD and its associated complications through the medium of scientific research, reviews and commentaries. Nonetheless, the molecular mechanisms underpinning ALD, particularly those underlying toxic effects of metabolic derivatives of alcohol on parenchymal and non-parenchymal hepatic cells leading to increased risk of alcohol-induced fibro-hepatocarcinogenesis, are still incompletely elucidated. In this review, we examined published scientific findings on how alcohol and its metabolic derivatives mount cellular attack on each hepatic cell and the underlying molecular mechanisms leading to disruption of core hepatic homeostatic functions which probably set the stage for the initiation and progression of ALD to fibro-hepatocarcinogenesis. We also brought to sharp focus, the complex and integrative role of transforming growth factor beta/small mothers against decapentaplegic/plasminogen activator inhibitor-1 and the mitogen activated protein kinase

Topics: Animals; Carcinogenesis; Carcinoma, Hepatocellular; Ethanol; Gastrointestinal Microbiome; Humans; Liver; Liver Cirrhosis, Alcoholic; Liver Diseases, Alcoholic; Liver Neoplasms; MAP Kinase Signaling System; Plasminogen Activator Inhibitor 1; Signal Transduction; Transforming Growth Factor beta

Transforming growth factor (TGF)-β is a pluripotent cytokine that displays several tissue-specific biological activities. In the liver, TGF-β is considered a fundamental molecule, controlling organ size and growth by limiting hepatocyte proliferation. It is involved in fibrogenesis and, therefore, in worsening liver damage, as well as in triggering the development of hepatocellular carcinoma (HCC). TGF-β is known to act as an oncosuppressor and also as a tumour promoter in HCC, but its role is still unclear.. In this review, we discuss the potential role of TGF-β in regulating the tumoural progression of HCC, and therefore the rationale for targeting this molecule in patients with HCC.. A considerable amount of experimental preclinical evidence suggests that TGF-β is a promising druggable target in patients with HCC. To support this hypothesis, a phase II clinical trial is currently ongoing using a TGF-β pathway inhibitor, and results will soon be available.. The identification of new TGF-β related biomarkers will help to select those patients most likely to benefit from therapy aimed at inhibiting the TGF-β pathway. New formulations that may provide a more controlled and sustained delivery of the drug will improve the therapeutic success of such treatments.

Topics: Carcinoma, Hepatocellular; Epithelial-Mesenchymal Transition; Humans; Liver Cirrhosis; Liver Diseases, Alcoholic; Liver Neoplasms; Signal Transduction; Transforming Growth Factor beta

Alcoholic liver disease remains a frequent and serious problem for increasing numbers of patients. Research has expanded our molecular understanding of the cellular basis of disease progression; however, translation into therapy is still hampered by a lack of suitable animal models for alcoholic liver disease, as well as from consequences of related liver damage due to malnutrition, hepatitis C virus infection, or abuse of other substances. Many patients with liver disease do not simply consume too much alcohol; they also suffer from comorbidities such as obesity or viral hepatitis, and/or may be addicted to other drugs besides alcohol. This review will summarize the currently available animal models to study liver disease due to either single causes or combinations of liver toxic substances/infections and alcohol.

Topics: Animals; Cytochrome P-450 CYP2E1; Disease Models, Animal; Hepatitis C, Chronic; Humans; Liver; Liver Diseases, Alcoholic; Malnutrition; Obesity, Morbid; Transforming Growth Factor beta

Alcoholic liver disease (ALD) remains a leading cause of death from liver disease in the United States for which there is no FDA-approved therapy. Abnormal cytokine metabolism is a major feature of ALD. Elevated serum concentration levels of TNF-alpha and TNF-alpha-inducible cytokines/chemokines, such as IL-6, -8, and -18, have been reported in patients with alcoholic hepatitis and/or cirrhosis, and levels correlated with markers of the acute phase response, liver function, and clinical outcome. Studies in animal models support an etiologic role for cytokines in the liver injury of ALD. Cytokines, such as transforming growth factor-beta, play a critical role in the fibrosis of ALD. Multiple new strategies are under investigation to modulate cytokine metabolism as a form of therapy for ALD.

Topics: Animals; Cytokines; Humans; Liver; Liver Cirrhosis; Liver Diseases, Alcoholic; Liver Regeneration; Transforming Growth Factor beta

Chronic alcoholism is a disease, which is known to affect individual's resistance to infections. Alcohol has a variety of short- and long-term effects on cell mediated and humoral immune response. Alcohol also affects the production, site of action and metabolism of cytokines, which are regulatory peptides of the immune and inflammatory response of lymphocytes, monocytes/macrophages and a variety of other cell types. In this review, current knowledge regarding the effect of alcohol on the immune system and the release and action of cytokines in humans and in animal models, in vivo and in vitro, is presenting. Furthermore, the role of cytokines in development of alcoholic liver disease is also discussed.

Topics: Animals; Antibody Formation; Autoimmune Diseases; Cytokines; Ethanol; Humans; Immunity, Cellular; Interferons; Interleukins; Lipopolysaccharides; Liver; Liver Diseases, Alcoholic; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

The pathogenesis of primary biliary cirrhosis (PBC) is unknown. The role of cytokines such as tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta (TGF-beta), and the effect of ursodeoxycholic acid (UDCA) in modifying the cytokine environment in patients with PBC has remained largely unstudied. Our aims were to determine: (i) the relationship between serum levels of TNF-alpha and TGF-beta and the severity of PBC; and (ii) the effects of UDCA therapy on TNF-alpha and TGF-beta levels in patients with PBC.. We studied 90 patients who had been treated with UDCA (53 patients) or placebo (37 patients) for 2 years as part of a randomized, double-blind, controlled trial. Patients were divided into histological stage I/II or stage III/IV disease. Serum TNF-alpha and TGF-beta levels were quantified by enzyme-linked immunoabsorbent assay.. Baseline levels of TNF-alpha were significantly greater in patients with stage III/IV compared to stage I/II disease. After 2 years of treatment with UDCA, patients showed a significantly greater decrease in TNF-alpha levels and progression risk score compared to placebo-treated patients. TNF-alpha and TGF-beta levels were significantly reduced compared to baseline levels in the UDCA-treated group after 2 years, while there was no significant change in the levels of placebo-treated patients.. Serum TNF-alpha and TGF-beta levels may reflect severity of disease in patients with PBC. The beneficial effects of UDCA therapy may be explained by lowering serum levels of these two cytokines.

Topics: Adult; Biomarkers; Biopsy; Cholagogues and Choleretics; Cholangitis, Sclerosing; Double-Blind Method; Female; Hepatitis C; Humans; Liver; Liver Cirrhosis, Biliary; Liver Diseases, Alcoholic; Male; Middle Aged; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Ursodeoxycholic Acid

Impaired liver regeneration has been considered as a hallmark of progression of alcohol-associated liver disease. Our previous studies demonstrated that in vivo inhibition of the microRNA (miRNA) miR21 can restore regenerative capacity of the liver in chronic ethanol-fed animals. The present study focuses on the role of microRNA regulatory networks that are likely to mediate the miR-21 action. Rats were chronically fed an ethanol-enriched diet along with pair-fed control animals and treated with AM21 (anti-miR-21), a locked nucleic acid antisense to miR-21. Partial hepatectomy (PHx) was performed and miRNA expression profiling over the course of liver regeneration was assessed. Our results showed dynamic expression changes in several miRNAs after PHx, notably with altered miRNA expression profiles between ethanol and control groups. We found that in vivo inhibition of miR-21 led to correlated differential expression of miR-340-5p and anticorrelated expression of miR-365, let-7a, miR-1224, and miR-146a across all sample groups after PHx. Gene set enrichment analysis identified a miRNA signature significantly associated with hepatic stellate cell activation within whole liver tissue data. We hypothesized that at least part of the PHx-induced miRNA network changes responsive to miR-21 inhibition is localized to hepatic stellate cells. We validated this hypothesis using AM21 and TGF-β treatments in LX-2 human hepatic stellate cells in culture and measured expression levels of select miRNAs by quantitative RT-PCR. Based on the in vivo and in vitro results, we propose a hepatic stellate cell miRNA regulatory network as contributing to the restoration of liver regenerative capacity by miR-21 inhibition.

Topics: Alcohol Drinking; Animals; Cell Line; Diet; Disease Models, Animal; Ethanol; Gene Regulatory Networks; Hepatectomy; Hepatic Stellate Cells; Humans; Liver Diseases, Alcoholic; Liver Regeneration; Male; MicroRNAs; Oligonucleotides; Rats; Rats, Sprague-Dawley; Signal Transduction; Transcriptome; Transfection; Transforming Growth Factor beta

Topics: 5'-Nucleotidase; Acetaldehyde; Adenosine; Adenosine Triphosphate; Animals; Antigens, CD; Apyrase; Carbon Tetrachloride; Colchicine; Cytokines; Disease Models, Animal; Ethanol; Gene Knockdown Techniques; Hepatic Stellate Cells; Humans; Liver Diseases, Alcoholic; Mice, Inbred C57BL; Primary Cell Culture; Rats; Receptor, Adenosine A2A; Receptor, Adenosine A2B; Signal Transduction; Smad3 Protein; Transforming Growth Factor beta; Tungsten Compounds

The let-7/Lin28 axis is associated with the regulation of key cellular regulatory genes known as microRNAs in various human disorders and cancer development. This study evaluated the role of the let-7/Lin28 axis in regulating a mesenchymal phenotype of hepatic stellate cells in alcoholic liver injury. We identified that ethanol feeding significantly down-regulated several members of the let-7 family in mouse liver, including let-7a and let-7b. Similarly, the treatment of human hepatic stellate cells (HSCs) with lipopolysaccharide (LPS) and transforming growth factor-β (TGF-β) significantly decreased the expressions of let-7a and let-7b. Conversely, overexpression of let-7a and let-7b suppressed the myofibroblastic activation of cultured human HSCs induced by LPS and TGF-β, as evidenced by repressed ACTA2 (α-actin 2), COL1A1 (collagen 1A1), TIMP1 (TIMP metallopeptidase inhibitor 1), and FN1 (fibronectin 1); this supports the notion that HSC activation is controlled by let-7. A combination of bioinformatics, dual-luciferase reporter assay, and Western blot analysis revealed that Lin28B and high-mobility group AT-hook (HMGA2) were the direct targets of let-7a and let-7b. Furthermore, Lin28B deficiency increased the expression of let-7a/let-7b as well as reduced HSC activation and liver fibrosis in mice with alcoholic liver injury. This feedback regulation of let-7 by Lin28B is verified in hepatic stellate cells isolated by laser capture microdissection from the model. The identification of the let-7/Lin28 axis as an important regulator of HSC activation as well as its upstream modulators and down-stream targets will provide insights into the involvement of altered microRNA expression in contributing to the pathogenesis of alcoholic liver fibrosis and novel therapeutic approaches for human alcoholic liver diseases.

Topics: Actins; Animals; Collagen Type I; Collagen Type I, alpha 1 Chain; DNA-Binding Proteins; Hepatic Stellate Cells; Humans; Lipopolysaccharides; Liver; Liver Diseases, Alcoholic; Mice; MicroRNAs; RNA-Binding Proteins; Signal Transduction; Transforming Growth Factor beta

To test the significance of lipid peroxidation in the development of alcoholic liver injury, an ethanol (EtOH) liquid diet was fed to male 129/SvJ mice (wild-type, WT) and glutathione S-transferase A4-4-null (GSTA4-/-) mice for 40 days. GSTA4-/- mice were crossed with peroxisome proliferator-activated receptor-α-null mice (PPAR-α-/-), and the effects of EtOH in the resulting double knockout (dKO) mice were compared with the other strains. EtOH increased lipid peroxidation in all except WT mice (P < 0.05). Increased steatosis and mRNA expression of the inflammatory markers CXCL2, tumor necrosis factor-α (TNF-α), and α-smooth muscle actin (α-SMA) were observed in EtOH GSTA4-/- compared with EtOH WT mice (P < 0.05). EtOH PPAR-α-/- mice had increased steatosis, serum alanine aminotransferase (ALT), and hepatic CD3+ T cell populations and elevated mRNA encoding CD14, CXCL2, TNF-α, IL-6, CD138, transforming growth factor-β, platelet-derived growth factor receptor-β (PDGFR-β), matrix metalloproteinase (MMP)-9, MMP-13, α-SMA, and collagen type 1 compared with EtOH WT mice. EtOH-fed dKO mice displayed elevation of periportal hepatic 4-hydroxynonenal adducts and serum antibodies against malondialdehyde adducts compared with EtOH feeding of GSTA4-/-, PPAR-α-/-, and WT mice (P < 0.05). ALT was higher in EtOH dKO mice compared with all other groups (P < 0.001). EtOH-fed dKO mice displayed elevated mRNAs for TNF-α and CD14, histological evidence of fibrosis, and increased PDGFR, MMP-9, and MMP-13 mRNAs compared with the EtOH GSTA4-/- or EtOH PPAR-α-/- genotype (P < 0.05). These findings demonstrate the central role lipid peroxidation plays in mediating progression of alcohol-induced necroinflammatory liver injury, stellate cell activation, matrix remodeling, and fibrosis.

Topics: Actins; Alanine Transaminase; Aldehydes; Animals; Antibodies; Chemokine CXCL2; Cytokines; Fibrosis; Gene Deletion; Glutathione Transferase; Lipid Peroxidation; Lipopolysaccharide Receptors; Liver; Liver Diseases, Alcoholic; Male; Matrix Metalloproteinases; Mice; PPAR alpha; Receptor, Platelet-Derived Growth Factor beta; RNA, Messenger; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

To evaluate protective effects of Chunggan extract (CGX), a traditional herbal formula, under 4 wk of alcohol consumption-induced liver injury.. Male Sprague-Dawley Rats were orally administered 30% ethanol daily for 4 wk with or without CGX. The pharmaceutical properties were assessed through liver enzymes, histopathology, fibrogenic cytokines, and alcohol metabolism in hepatic tissues as well as by in vitro experiment using HSC-T6 cells.. Four weeks of alcohol consumption notably increased liver enzymes and malondialdehyde levels in serum and hepatic tissue. CGX not only prevented the collagen deposition determined by histopathology and hydroxyproline content, but also normalized transforming growth factor-beta, platelet-derived growth factor-beta and connective tissue growth factor at the gene expression and protein levels in liver tissue. Moreover, CGX treatment also significantly normalized the abnormal changes in gene expression profiles of extracellular matrix proteins, matrix metalloproteinase and their inhibitors, alcohol metabolism, and inflammatory reactions. In the acetaldehyde-stimulated HSC-T6 cells, CGX considerably inhibited collagen production and normalized fibrogenic cytokines in both gene expression and protein levels.. The present study evidenced that CGX has hepatoprotective properties via modulation of fibrogenic cytokines and alcohol metabolism in alcoholic liver injury.

Topics: Animals; Cell Line; Collagen; Connective Tissue Growth Factor; Cytokines; Disease Models, Animal; Drugs, Chinese Herbal; Gene Expression Regulation; Hepatocytes; Hydroxyproline; Liver; Liver Diseases, Alcoholic; Male; Malondialdehyde; Phytotherapy; Plants, Medicinal; Protective Agents; Proto-Oncogene Proteins c-sis; Rats, Sprague-Dawley; RNA, Messenger; Time Factors; Transforming Growth Factor beta

Curcumin is known for its antiinflammatory and antifibrotic properties in liver damage. However, the negative effects of curcumin on alcoholic liver damage are seldom reported. In this study, both advantageous and disadvantageous functions of curcumin on alcoholic liver injury were observed. In order to determine the effects of curcumin on liver fibrosis induced by alcohol, 5% ethanol and/or curcumin (1 × 10(-3) or 1 × 10(-4)  m) were injected intravenously in mice. Hematoxylin-eosin staining was performed to determine the value of liver injury by optical density analyses. Liver histology was evaluated by an experienced hepatopathologist blinded to the type of treatment received by the animals. Ethanol accelerates serum levels of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT), liver injury, production of tumor necrosis factor-alpha (TNF-α), transforming growth factor-beta (TGF-β) and nuclear factor-kappa B (NF-κB) during 5% ethanol-induced liver injury. 1 × 10(-3)  m curcumin accelerates liver injury and liver cellular edema during only 5% ethanol-induced liver injury evolution, whereas 1 × 10(-4)  m curcumin does not lead to (or protect) alcoholic liver injury. Therefore, it is suggested that curcumin may have dual effects on alcoholic liver injury depending on its concentration.

Topics: Alanine Transaminase; Animals; Aspartate Aminotransferases; Curcumin; Dose-Response Relationship, Drug; Ethanol; Liver; Liver Diseases, Alcoholic; Male; Mice; Mice, Inbred C57BL; NF-kappa B; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Quantification of hepatocyte death is useful to evaluate the progression of alcoholic liver diseases. Our aims were to quantify and correlate the circulating levels of Cytokeratin 18 (CK18) and its caspases-generated fragment to disease severity in heavy alcoholics.. CK18 and CK18-fragment were evaluated in the serum of 143 heavy alcoholics. Serum levels of markers of hepatocyte death (CK18), apoptosis (CK18 fragment) and necrosis (CK18 -CK18 fragment) increased in patients with severe fibrosis compared to patients with mild fibrosis. These markers strongly correlated with Mallory-Denk bodies, hepatocyte ballooning, fibrosis and with hepatic TNFα and TGFβ assessed in the liver of 24 patients. Elevated levels of serum hepatocyte death and apoptotic markers were independent risk factors in predicting severe fibrosis in a model combining alkaline phosphatase, bilirubin, prothrombin index, hyaluronate, hepatocyte death and apoptotic markers. The level of markers of hepatocyte death and apoptosis had an area under the receiving operator curve that predicted severe fibrosis of 0.84 and 0.76, respectively.. Death of hepatocytes can be easily evaluated with serum markers and correlated with severe fibrosis in heavy alcohol drinkers. These biomarkers could be useful to rapidly evaluate liver injuries and the efficacy of therapies.

Topics: Adult; Apoptosis; Biomarkers; Female; Hepatocytes; Humans; Liver Cirrhosis; Liver Diseases, Alcoholic; Male; Middle Aged; Polymerase Chain Reaction; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Alcoholic liver disease is continuously increasing in developed countries being a leading cause of death worldwide. Chronic ethanol consumption induces oxidative stress by accumulation of reactive oxygen intermediates (ROI) while reducing the cellular antioxidant defense. Induction of heme oxygenase-1 (HO-1) may protect primary human hepatocytes (hHeps) from such damage. Thus, the aim of this study was to investigate the potential of polyphenols to protect hHeps from ethanol-dependent oxidative damage.. hHeps were isolated by collagenase perfusion. ROI and cellular glutathione (GSH) were measured by fluorescent-based assays. Cellular damage was determined by lactate dehydrogenase (LDH) leakage and staining for apoptosis and necrosis. Nuclear translocation of Nrf2 and HO-1 expression were analyzed by Western blot.. Ethanol and TGF-β rapidly increase ROI and reduce GSH in hHeps, causing apoptosis with a release of approximately 40% total LDH after 72 h. Similar to incubation with hemin preincubation and co-incubation of cells with nifedipine, verapamil and quercetin significantly reduce oxidative stress and resulting cellular damage, in a dose-dependent manner, by initiating nuclear translocation of Nrf2 which in turn induces HO-1 under the control of p38 and ERK. Blocking of HO-1 activity with ZNPP9 reverses the protective effect of all three substances.. Our results suggest that increasing HO-1 activity in hHeps protects them from oxidative stress-dependent damage. As polyphenols have great potential to induce HO-1 expression, they may play an important role for future therapeutic strategies to protect liver from oxidative stress-dependent damage observed during chronic alcohol consumption.

Topics: Buffers; Cytoprotection; Ethanol; Flavonoids; Heme Oxygenase-1; Hepatocytes; Humans; L-Lactate Dehydrogenase; Liver Diseases, Alcoholic; Models, Biological; Oxidative Stress; Phenols; Polyphenols; Protective Agents; Transforming Growth Factor beta; Up-Regulation

Topics: Alcohol-Related Disorders; Animals; Antioxidants; Cytokines; Ethanol; Humans; Liver; Liver Cirrhosis, Alcoholic; Liver Diseases, Alcoholic; Oxidative Stress; Pancreas; Pancreatitis, Alcoholic; Signal Transduction; Transforming Growth Factor beta

There are wide interindividual differences in the risk of developing alcoholic cirrhosis. Transforming growth factor beta(1) (TGF-beta(1)) is the main cytokine involved in liver fibrogenesis. The TGF-beta(1) gene is polymorphic at several sites and these polymorphisms are probably related to differences in the rate of TGF-beta(1) synthesis. Our aim has been to analyse the influence of the TGF-beta(1) gene polymorphisms in the predisposition to advanced alcoholic liver disease (ALD) in ethanol abusers.. TGF-beta(1) single nucleotide polymorphisms at positions -509 (C or T), +869 (C or T, codon 10), and +915 (C or G, codon 25) were examined in 165 alcoholics with advanced ALD and in 185 healthy controls.. Among the 94 male patients with oesophageal varices, those carrying the GG genotype at position +915 were diagnosed at an older age than the remaining patients (age 52.1 years, standard deviation (SD) 9.9 vs. 45 SD 13.4, P=0.012). No other statistically significant differences were found in the distribution of the three TGF-beta(1) polymorphisms analysed individually or as combined haplotypes.. The polymorphisms at the TGF-beta(1) gene analysed in this study are probably not related to the risk of advanced ALD.

Topics: Adult; Aged; Aged, 80 and over; Female; Genotype; Haplotypes; Humans; Liver Diseases, Alcoholic; Male; Middle Aged; Polymorphism, Genetic; Sex Characteristics; Transforming Growth Factor beta; Transforming Growth Factor beta1

Topics: Humans; Leptin; Liver Cirrhosis; Liver Diseases, Alcoholic; Male; Transforming Growth Factor beta; Transforming Growth Factor beta1

Alcohol consumption accelerates the appearance of liver fibrosis and hepatocellular carcinoma in patients with chronic hepatitis C virus (HCV) infection, but the mechanisms of these interactions are unknown. We therefore investigated the effects of chronic ethanol consumption in HCV core protein-expressing transgenic mice.. Ethanol was progressively added (up to 20%) to the drinking water that was given ad libidum.. In vivo fatty acid oxidation was not inhibited by ethanol consumption and/or HCV core expression. Both chronic ethanol consumption and HCV core expression decreased hepatic lipoprotein secretion and caused steatosis, but had no additive effects on lipoprotein secretion or steatosis. However, chronic ethanol consumption and HCV core protein additively increased lipid peroxidation and acted synergistically to increase the hepatic expression of transforming growth factor-beta (TGF-beta) and, to a less extent, tumor necrosis factor-alpha (TNF-alpha).. HCV core protein expression and chronic alcohol consumption have no effects on in vivo fatty acid oxidation and do not additively impair hepatic lipoprotein secretion, but additively increase hepatic lipid peroxidation and synergistically increase hepatic TNF-alpha and TGF-beta expression. These effects may be involved in the activation of fibrogenesis and the development of hepatocellular carcinoma in patients cumulating alcohol abuse and HCV infection.

Topics: Animals; Central Nervous System Depressants; Disease Models, Animal; Ethanol; Fatty Acids; Fatty Liver; Gene Expression; Hepacivirus; Hepatitis B Core Antigens; Lipid Peroxidation; Lipoproteins; Liver; Liver Diseases, Alcoholic; Mice; Mice, Inbred C57BL; Mice, Transgenic; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Topics: Alanine Transaminase; Animals; Aspartate Aminotransferases; Cytokines; Ethanol; Gene Expression Regulation; Interleukin-18; Liver Diseases, Alcoholic; Male; Platelet-Derived Growth Factor; Rats; Rats, Wistar; Transforming Growth Factor beta; Triglycerides

To investigate the expression of the transforming growth factor beta 1(TGF-beta 1) mRNA in different stages of alcoholic liver disease (ALD) and its clinical value.. One hundred and seven male alcoholics were grouped by clinical findings into four groups: alcohol abusers without liver impairment (n =22), alcoholic steatosis (n =30); alcoholic hepatitis (n=31); and alcoholic cirrhosis(n=24). Using peripheral blood mononuclear cells (PBMC) as samples the gene expression of TGF-beta 1 was examined quantitatively by reverse transcription polymerase chain reaction (RT-PCR) and dot blot. There are 34 healthy subjects served as control.. The expression of TGF-beta 1 from all ALD patients was significantly greater than that in controls (1.320 +/- 1.162 vs 0.808 +/- 0.276, P<0.001). The differences of the expressions were significant between the patients from each groups (alcoholic steatosis, alcoholic hepatitis and alcoholic cirrhosis) and the controls (1.168 +/- 0.852, 1.462 +/- 1.657, 1.329 +/- 0.610 vs 0.808 +/- 0.276, P<0.050). No significant differences of TGF -beta 1 mRNA expression were observed between alcohol abusers without liver impairment and controls. The expressions in patients with alcoholic hepatitis and alcoholic cirrhosis were significantly greater than that in alcohol abusers respectively (1.462 +/- 1.657, 1.329 +/- 0.610 vs 0.841 +/- 0.706, P<0.050). No significant differences of TGF-beta 1 mRNA expression were observed between alcoholic fatty liver men and alcohol abusers.. TGF-beta 1 expression level can be a risk factor for alcoholic liver disease and might be related to the inflammatory activity and fibrosis of the liver in patients.

Topics: Humans; Liver Diseases, Alcoholic; Male; RNA, Messenger; Transforming Growth Factor beta; Transforming Growth Factor beta1

To determine what changes are occurring in patients with primary sclerosing cholangitis (PSC) by examining perisinusoidal macrophages (Kupffer cells) in liver biopsies; 2-to measure transforming growth factor beta (TGFbeta) as a marker of fibrosis in these patients.. Transmission electron microscopy and immunohistochemistry of 15 PSC, 26 primary biliary cirrhosis (PBC), 30 alcoholic liver disease (ALD) and 51 with normal histology was used. Five PSC, 30 ALD and 120 normal volunteers were sampled for serum levels of TGFbeta.. There was a three-fold increase in relative numbers of Kupffer cells in PSC compared to PBC and to patients whose livers had normal histology. In PSC there was an accumulation of perisinusoidal macrophages, which was not associated with focal necrosis or with cholestasis. The levels of TGFbeta in PSC were 54 +/- 2 in cirrhotic versus 34 +/- 5 in non-cirrhotic patients (p < 0.005).. The persistent activation of these macrophages may lead to the chronic release of TGFbeta and contribute to chronic inflammation, fibrosis and cirrhosis.

Topics: Adolescent; Adult; Aged; Biopsy; Case-Control Studies; Cholangitis, Sclerosing; Female; Humans; Immunohistochemistry; Liver; Liver Cirrhosis, Biliary; Liver Diseases, Alcoholic; Macrophages; Male; Microscopy, Electron; Middle Aged; Transforming Growth Factor beta

Hepatic fibrosis in alcoholic liver disease often heralds progression to cirrhosis and, therefore, noninvasive parameters are required for early diagnosis and follow-up. Collagens VI and XIV, procollagen-III-N-propeptide, hyaluronic acid, and active transforming growth factor-beta1 (TGF-beta1) were measured in healthy volunteers, patients with alcoholic cirrhosis, and heavy drinkers without cirrhosis. Noncirrhotic alcoholics were assigned to two groups with either normal aspartate aminotransferase or levels > or = 2 normal. Collagens VI and XIV were elevated in all alcoholic patients compared to controls (P < 0.0001, all instances). Procollagen-III-N-propeptide and hyaluronic acid levels were higher in alcoholic patients with elevated liver enzymes and in cirrhotics as compared to controls. Procollagen-III-N-propeptide revealed a significant correlation with serum levels of TGF-beta1 (P < 0.0001). Collagens VI, and XIV, procollagen-III-N-propeptide, and hyaluronic acid appear to be sensitive markers indicating fibrotic transformation in alcoholics. The correlation between procollagen-III-N-propeptide and TGF-beta1 emphasizes its role in hepatic fibrogenesis.

Topics: Adult; Alcoholism; Biomarkers; Collagen; Connective Tissue; Female; Humans; Hyaluronic Acid; Liver Cirrhosis, Alcoholic; Liver Diseases, Alcoholic; Male; Peptide Fragments; Procollagen; Transforming Growth Factor beta; Transforming Growth Factor beta1

Many acute and chronic liver diseases are often associated with atypical ductular proliferation (ADP). These ADPs have gained increasing interest since a number of recent observations suggest that ADPs may represent progenies of the putative liver stem cell compartment. In this study, we show that feeding mice with 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) results in persistent proliferation of primitive ductules with poorly defined lumens. Similar to oval cell proliferation in other rodent models as well as in various human liver diseases, DDC-induced ADP originated from the portal tract, spread into the hepatic lobule, and was associated closely with appearance of hepatocytes harboring an antigen (A6), which normally is expressed in biliary epithelium. Furthermore, DDC treatment severely inhibited the regenerative capacity of mice after partial hepatectomy. The development of ADP was selectively blocked in DDC-fed TGF-beta1 transgenic mice producing active TGF-beta1 in the liver and no accumulation of new hepatocytes expressing the A6 antigen was observed. Moreover, the transforming growth factor beta1 (TGF-beta1) transgenic mice did not survive beyond 3 weeks from starting the DDC-containing diet. The results suggest that persistent activation of the hepatic stem cell compartment is essential for liver regeneration in the DDC model and that active TGF-beta1 may negatively control activation of stem cells in the liver. These data further emphasize the relevance of the DDC model as an experimental tool for studying chronic liver diseases.

Topics: Animals; Bile Ducts, Intrahepatic; Cell Division; Chemical and Drug Induced Liver Injury; Chronic Disease; Common Bile Duct; Dicarbethoxydihydrocollidine; Disease Models, Animal; Epithelial Cells; Hepatectomy; Ligation; Liver; Liver Diseases, Alcoholic; Liver Regeneration; Mice; Mice, Transgenic; Transforming Growth Factor beta

The release of proinflammatory cytokines by endotoxins and during oxidative stress is considered to be an early key step in the pathogenesis of alcoholic liver disease (ALD). Ethanol-inducible cytochrome P450 2E1 (CYP2E1) has potentially pro-oxidative and toxicological properties, and its expression is restricted to the perivenous region of liver. We investigated zonal differences of cytokine expression in rat liver and how these are affected by alcohol exposure and by chlormethiazole (CMZ), a transcriptional and posttranslational inhibitor of hepatic CYP2E1. Periportal and perivenous cell lysates were obtained by the digitonin pulse technique from livers of rats treated with ethanol and CMZ for 38 days. Cytokine expression on the mRNA and protein levels was quantified using competitive polymerase chain reaction (PCR) and Western blot, respectively. Chronic ethanol treatment significantly increased the expression of CYP2E1, microsomal p-nitrophenol hydroxylase activity (indicative for CYP2E1 enzyme activity), and the expression of transforming growth factor beta1 (TGF-beta1), tumor necrosis factor alpha (TNF-alpha), and interleukin (IL)-1beta (1.4- to 4.6-fold). In contrast, ethanol caused a decrease in IL-4 expression and had no influence on IL-6 expression. CMZ treatment caused a reduction in hepatic CYP2E1 expression and in the ethanol-induced cytokine expression by 40% to 60%. Expression of IL-6, IL-2, and IL-4 mRNA occurred preferentially in the periportal region, whereas ethanol caused a pronounced increase in the perivenous expression of TGF-beta1, which was inhibited by CMZ as monitored both on the mRNA and protein levels. These results show the zonated expression of several cytokines and the counteraction of CMZ on all effects of ethanol on cytokine expression. The data further strengthen a link between increased CYP2E1 expression and enhanced cytokine expression as important events in the development of ALD.

Topics: Animals; Chlormethiazole; Cytochrome P-450 CYP2E1 Inhibitors; Cytokines; Ethanol; Gene Expression; Interleukin-1; Interleukins; Liver; Liver Diseases, Alcoholic; Male; Rats; Rats, Sprague-Dawley; Rats, Wistar; RNA, Messenger; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Thromboxane levels correlate with severity of liver injury in rats given alcohol. The aim of this study was to evaluate the effect of thromboxane inhibitors on pathological changes in experimental alcoholic liver disease.. Male Wistar rats were given a liquid diet and ethanol intragastrically for 1 month. The thromboxane inhibitors tested were a thromboxane receptor antagonist (TXRA) and a thromboxane synthase inhibitor (TXSI). Pathological changes, liver and plasma thromboxane levels, 6-ketoprostaglandin F1 alpha levels, lipid peroxidation, and messenger RNA levels for tumor necrosis factor (TNF)-alpha and transforming growth factor (TGF) beta were evaluated.. Treatment with thromboxane inhibitors prevented necrosis and inflammation. In the TXSI-treated group, fatty liver was also decreased. Ethanol administration led to a 3-4-fold increase in liver thromboxane levels; a reduction in thromboxane levels and lipid peroxidation was seen in the TXSI group. In all treatment groups, TNF-alpha and TGF-beta messenger RNA levels were decreased.. The prevention of necroinflammatory changes in thromboxane-treated groups is related to a decrease in TNF-alpha levels. Inhibition of TGF-beta expression may also prevent fibrosis in ethanol-treated rats.

Topics: Animals; Bridged Bicyclo Compounds, Heterocyclic; Fatty Liver, Alcoholic; Liver Diseases, Alcoholic; Male; Oxazoles; Rats; Rats, Wistar; Thromboxane A2; Thromboxane B2; Transforming Growth Factor alpha; Transforming Growth Factor beta

Based on studies that show a role for the low-density lipoprotein (LDL)-receptor in arachidonic acid delivery and eicosanoid synthesis in macrophages, the present study investigated the effect of cholesterol supplementation on pathological changes and thromboxane (TX) synthesis in alcoholic liver injury. Male Wistar rats were intragastrically fed ethanol with either corn oil or fish oil for 1 month. Control rats received isocaloric amounts of dextrose instead of ethanol. An additional group of rats fed either ethanol or dextrose with fish oil or corn oil were supplemented with 1% cholesterol. At the time of killing, all rats had the following evaluated: liver histopathology, lipid peroxidation, liver and plasma thromboxane levels, plasma endotoxin and messenger RNA (mRNA) levels of LDL-receptor, tumor necrosis factor alpha (TNF-alpha), cyclooxygenase (Cox)-1 and -2, and transforming growth factor beta (TGF-beta). Rats fed ethanol with either fish oil or corn oil developed fatty liver, necrosis, inflammation, and central vein collagen deposition. Cholesterol supplementation enhanced the degree of fibrosis but prevented necrosis and inflammation. These alterations in pathological changes by cholesterol were accompanied by absent TNF-alpha and Cox-2 mRNAs, decreased thromboxane levels, decreased lipid peroxidation, and increased TGF-beta mRNA. Cholesterol enrichment of the diet thus decreases proinflammatory components, but enhances fibrosis in ethanol-fed rats.

Topics: Animals; Cholesterol; Inflammation; Lipids; Liver; Liver Cirrhosis; Liver Diseases, Alcoholic; Male; Necrosis; Rats; Rats, Wistar; RNA, Messenger; Thromboxane B2; Transforming Growth Factor beta

We used the intragastric feeding rat model for alcoholic liver disease to investigate the relationship between transforming growth factor (TGF)-beta 1 and inhibition of endothelial cell proliferation. Twelve groups of male Wistar rats (four to five rats per group) were fed ethanol or dextrose with either corn oil or saturated fat for 1-, 2-, and 4-week periods. All control animals were pair fed the same diets as ethanol-fed rats except that ethanol was isocalorically replaced by dextrose. In the ethanol-fed groups, nonparenchymal cells were isolated and TGF-beta 1 was measured in the nonparenchymal cell supernatant. Liver pathology and endothelial cell proliferation with an antibody to proliferating cell nuclear antigen were studied in all groups. Plasma TGF-beta 1 was measured in all rats. Pathological changes (fatty liver, necrosis, and inflammation) were observed only in the corn oil/ethanol-fed rats at 4 weeks. Significantly higher levels of TGF-beta 1 were seen in both plasma and nonparenchymal cell supernatant in rats fed corn oil and ethanol; plasma levels of TGF-beta 1 were not significantly different between the dextrose-fed controls and saturated fat/ethanol-fed rats. A significant inverse correlation (r = -0.89, P < 0.01) was seen between plasma TGF-beta 1 and the number of endothelial cells arrested at G1/S. Immunohistochemistry revealed the presence of TGF-beta 1 staining in interstitial macrophages only in rats fed corn oil and ethanol. The present study provides evidence for a role for TGF-beta 1 in inhibiting endothelial cell proliferation in experimental alcoholic liver disease. Arrest of endothelial cells may lead to their differentiation and/or to produce mediators that could stimulate other cells such as Ito cells. Sustained TGF-beta 1 may also lead to Ito cell production of extracellular matrix.

Topics: Animals; Cell Division; Endothelium, Vascular; Immunohistochemistry; Liver; Liver Diseases, Alcoholic; Male; Rats; Rats, Wistar; Transforming Growth Factor beta; Vimentin

Kupffer cell-derived cytokines are believed to play pivotal paracrine roles in the pathogenesis of alcoholic liver disease (ALD). To evaluate this hypothesis, Kupffer cell gene expression of tumor necrosis factor-alpha (TNF alpha), interleukin (IL)-6, and transforming growth factor-beta 1 (TGF beta 1) were directly examined in the rat model of ALD. Kupffer cells were isolated from the model after 10 and 17 weeks of intragastric ethanol infusion. These two durations resulted in focal hepatocellular injury and liver fibrogenesis, respectively. Oxidative stress as assessed by the hepatic level of thiobarbituric acid reacting substances, was evident at 10 weeks but more pronounced at 17 weeks. The steady state messenger RNA (mRNA) levels of the cytokines were examined by Northern blot analysis using RNA samples from freshly isolated Kupffer cells, and the release of the cytokines was quantitated ex vivo using a 3-day culture. The mRNA levels of TNF alpha and TGF beta 1 were significantly increased by 183% and 204% at 10 weeks and 231% and 295% at 17 weeks in the ethanol-fed rats, respectively. Ex vivo release of TNF activity by control Kupffer cells was undetectable or very low (< 2U/10(5) cells/18 hours) at both time points, but the cells from the ethanol-fed animals secreted appreciably more TNF (27.8 +/- 7.6 U at 10 weeks and 40.4 +/- 10.3 U at 17 weeks). The release of the latent TGF beta 1 protein was also coordinately increased by 143% at 10 weeks and 238% at 17 weeks.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Alanine Transaminase; Animals; Aspartate Aminotransferases; Cytokines; Disease Models, Animal; Gene Expression; Interleukin-6; Kupffer Cells; Liver; Liver Diseases, Alcoholic; Male; Oxidative Stress; Rats; Rats, Wistar; RNA, Messenger; Thiobarbituric Acid Reactive Substances; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

We evaluated the expression of interleukin-1 alpha, interleukin-1 beta, tumor necrosis factor-alpha and transforming growth factor-beta mRNAs in the intragastric-feeding rat model of alcoholic liver disease. Rats were fed different diets for periods of 2 or 4 wk. Animals fed saturated fat and ethanol and the corn oil-dextrose control group had no liver injury, whereas animals fed corn oil and ethanol showed pathologic changes. RNA was extracted from the livers at the time of killing, reverse-transcribed and amplified; polymerase chain reaction products were subjected to electrophoresis on agarose gel. Interleukin-1 alpha mRNA was present in all groups at 2 and 4 wk; interleukin-1 beta and transforming growth factor-beta mRNAs were present in all groups at 4 wk. Tumor necrosis factor-alpha mRNA was absent in all groups at 2 wk but was present in the corn oil-ethanol group only at 4 wk. Because pathological liver injury was evident in the corn oil-ethanol group by 4 wk, the presence of tumor necrosis factor-alpha mRNA at this time suggests a pathogenetic role for tumor necrosis factor-alpha in alcohol-induced liver injury.

Topics: Animals; Base Sequence; Cytokines; Disease Models, Animal; DNA Primers; Gene Expression; Interleukin-1; Liver; Liver Diseases, Alcoholic; Male; Molecular Sequence Data; Polymerase Chain Reaction; Rats; Rats, Wistar; RNA, Messenger; Transcription, Genetic; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

In humans, alcoholic liver disease is frequently associated with IgA mesangial deposits, microscopic hematuria and a small amount of proteinuria, identifying a secondary form of IgA nephropathy. Alcoholic liver disease is almost always associated with nutritional deficiencies.. In order to examine the relationship between alcohol intake and/or inadequate diet and IgA nephropathy, groups of 4 week-old-male Lewis rats were maintained on a lipotrope-deficient (LD) diet (N = 20), intragastric infusions of a commercial whiskey (1.5 ml/100 gm body weight) three times a week, and regular chow (N = 23) or both intragastric whiskey infusion and an LD diet (N = 17). A fourth control group (N = 19) was given no whiskey and normal chow.. All rats given the LD diet had marked steatosis and elevated "liver" enzymes. Changes were more severe, and with early bridging fibrosis and nodule formation in those also given whiskey, associated with increased hepatic content of mRNA encoding transforming growth factor-beta. A moderate steatosis without alteration in serum enzymes or transforming growth factor-beta expression was found in rats given whiskey (all p < 0.0001) compared with controls. IgA accumulated in hepatic sinusoids instead of in canaliculi and bile ducts, suggesting impaired transport of IgA and IgA immune complexes from blood to bile, in rats given an LD diet and/or whiskey infusion. A moderate increase in mesangial matrix was observed only in rats given both whiskey and an LD diet. Bright granular IgA and mild granular C3 mesangial deposits and electron-dense deposits were evident in 63 to 70% of experimental rats (all p < 0.001) versus only trace deposits in 5 to 11% of controls. Moderate IgG codeposits were present in 34 to 55% of rats given the LD diet and/or whiskey (all p < 0.02), versus trace deposits in 10% of controls. Significant hematuria and proteinuria were observed in rats given the LD diet and/or whiskey (p < 0.0001) versus controls. Intestinal permeability measured by xylose absorption was significantly increased relative to controls only in rats given both whiskey and the LD diet (p < 0.001). Serum IgA specific for selected alimentary antigens was increased relative to controls in 75 to 100% of the experimental rats.. The combination of LD diet and alcohol intake, which mimics the human alcoholic condition, promotes hepatic and renal changes, leading to hepatocellular injury and a secondary form of IgA nephropathy.

Topics: Alcoholism; Animal Nutritional Physiological Phenomena; Animals; Deficiency Diseases; Disease Models, Animal; Fluorescent Antibody Technique; Glomerulonephritis, IGA; Immunoglobulin A; Intestinal Absorption; Kidney; Liver; Liver Diseases, Alcoholic; Male; Microscopy, Electron; Proteinuria; Rats; Rats, Inbred Lew; RNA, Messenger; Transforming Growth Factor beta; Xylose