Features Of Alcoholic Liver Disease Research Articles

Refining the Rab7-V1G1 axis to mitigate iron deposition: Protective effects of quercetin in alcoholic liver disease

Iron overload is a common feature of alcoholic liver disease (ALD) and contributes significantly to disease progression. Quercetin, a flavonoid known for its iron-chelating properties, has emerged as a potential protective compound against ALD. However, research on quercetin's regulatory effects on iron levels in ALD is limited. To address this, we conducted a study using male C57BL/6J mice were subjected to a Lieber De Carli liquid diet containing ethanol (28% energy replacement) with or without quercetin supplementation (100 mg/kg.BW) for 12 weeks. Additionally, HepG2 cells, after transfection with the CYP2E1 plasmid, were incubated with ethanol and/or quercetin. Our findings revealed that ethanol consumption led to iron overload in both hepatocytes and lysosomes. Interestingly, despite the increase in iron levels, cells exhibited impaired iron utilization, disrupting normal iron metabolism. Further analysis identified a potential mechanism involving the Rab7-V1G1 (V-ATPase subunit) axis. Inhibition of V-ATPase by Concanamycin A caused elevated ROS levels, impaired lysosomal and mitochondria function, and increased expression of HIF1α and IRP2. Ultimately, this disruption in cellular processes led to iron overload and mitochondrial iron deficiency. Quercetin supplementation mitigated ethanol-induced hepatocyte damage by reversing iron overload through modulation of the Rab7-V1G1 axis and improving the interaction between lysosomes and mitochondria. In conclusion, this study elucidates a novel pathophysiological mechanism by which quercetin protects against ALD through its regulation of iron homeostasis.

Pueraria lobata-Prunus mume Complex Alleviates Alcoholic Liver Disease by Regulating Lipid Metabolism and Inhibiting Inflammation: A Transcriptome and Gut Microbiota Analysis.

Lipid metabolism disorder appears to be one of the early features of alcoholic liver disease (ALD), which can be speculated via omics analysis including liver transcriptomics and gut microbiota. A complex consisting of the roots of Pueraria lobata and dried fruits of Prunus mume (PPC), which possesses hepatoprotective effects, could serve as a drug or functional food. The lack of non-polysaccharide compounds in PPC with their moderation effects on gut microbiota suggests the necessity for a relevant study. Six groups of Kunming mice (control, Baijiu injury, silybin, low, medium, and high) were modelled by gavage with Baijiu (for 14 days) and PPC (equivalent to a maximum dose of 9 g/kg in humans). The liver transcriptome data were analyzed to predict gene annotation, followed by the verification of gut microbiota, serum, tissue staining, immunohistochemistry, and Western blotting. Liquid chromatography-mass spectrometry was used to detect the components. PPC normalized serum ALT (40 U/L), down-regulated TLR4-NF-κB signaling pathway to inhibit the release of TNF-α (90 pg/mL), improved the expression of occludin, claudin-4, and ZO-1, and restored the abundance of Muribaculaceae, Bacteroides and Streptococcus. PPC can alleviate ALD by regulating the gut microbiota with an anti-inflammatory and intestinal barrier, and has an application value in developing functional foods.

Circulating cytokines and alcoholic liver disease: a two-sample bidirectional Mendelian randomization study

Background Increased inflammation in the liver during ethanol exposure is a major feature of alcoholic liver disease (ALD). An important contributing component to the development of ALD is the inflammatory response brought on by immunological response, however the connection between individual circulating cytokines and ALD is still unclear. To ascertain the causation, we conducted a two-sample bidirectional Mendelian randomization research. Methods We extracted 41 cytokines and growth factors of 8293 Europeans and ALD cases of the same ethnicity (1416 cases and 217,376 controls) from the Genome-Wide Association Studies (GWAS) database for two-sample bidirectional MR analysis. Results Our analyses suggest that higher interleukin-7 (IL-7) levels are associated with an increased risk of ALD (p = 0.028, OR = 1.191,95% CI = 1.019–1.392), while tumor necrosis factor related apoptosis inducing ligand (TRAIL) is a protective factor for ALD (p = 0.032, OR = 0.863, 95% CI = 0.754–0.988) which can reduce the risk of disease occurrence. In addition, genetically predicted ALD does not affect the expression of circulating cytokines regulators. Conclusions Our study supports that cytokines play a pivotal role in the pathogenesis of ALD. To determine the mechanisms and pathways of action of these biomarkers, further basic research is required to ensure their clinical suitability for preventing and treating ALD.

Metformin protects against ethanol-induced liver triglyceride accumulation by the LKB1/AMPK/ACC pathway.

Hepatic lipid accumulation is one of the main pathological features of alcoholic liver disease (ALD). Metformin serves as an AMPK activator and has been shown to have lipids lowering effects in non-alcoholic fatty liver disease (NAFLD), but its role in ALD remains unclear. The purpose of this study was to explore the potential mechanism of metformin regulating lipid metabolism in ALD. In vitro and in vivo ALD models were established using AML12 cells and C57BL/6 mice, respectively. To determine the effect of metformin on ALD in vitro, the concentration of cellular triglyceride was examined by Nile red staining and a biochemical kit. To further reveal the role of metformin on ALD in vivo, liver tissues were examined by HE and oil red O staining, and the levels of ALT and AST in serum were determined via an automatic biochemical analyzer. The expression of mRNA and proteins were measured using qRT-PCR and Western blot, respectively. The role of the LKB1/AMPK/ACC axis on metformin regulating ethanol-induced lipid accumulation was evaluated by siRNA and AAV-shRNA interference. The results showed metformin reduced the ethanol-induced lipid accumulation in AML12 cells through activating AMPK, inhibiting ACC, reducing SREBP1c, and increasing PPARα. In addition, compared with control mice, metformin treatment inhibited ethanol-induced liver triglyceride accumulation and the increase of ALT and AST in serum. Interference with LKB1 attenuated the effect of metformin on ethanol-induced lipid accumulation both in vitro and in vivo. Metformin protects against lipid formation in ALD by activating the LKB1/AMPK/ACC axis.

Nutritional Status and Diet Style Affect Cognitive Function in Alcoholic Liver Disease.

Malnutrition and cognitive dysfunction are typical features of alcoholic liver disease (ALD) and are correlated with the development of complications. The aim of this study is to explore the effect of nutritional state and diet on cognitive function in ALD. A total of 43 patients with compensated alcoholic cirrhosis were enrolled, and a neuropsychological test was assessed according to body mass index (BMI, <22 and ≥22). In the ALD animal study, mice were divided into five groups (n = 9/group; normal liquid, 5% EtOH + regular liquid, 5% EtOH + high-carbohydrate liquid, 5% EtOH + high-fat liquid, and 5% EtOH + high-protein liquid diet) and fed the same calories for eight weeks. To assess cognitive function, we performed T-maze studies weekly before/after alcohol binging. In cognitive function (BMI < 22/≥22), language score of Korea mini-mental state (7.4 ± 1.4/7.9 ± 0.4), Boston naming (11.7 ± 2.7/13.0 ± 1.8), forward digit span (6.7 ± 1.8/7.5 ± 1.6), Korean color word stroop (24.2 ± 26.5/43.6 ± 32.4), and interference score (33.9 ± 31.9/52.3 ± 33.9) revealed significant differences. In the T-maze test, alcohol significantly delayed the time to reach food, and binge drinking provided a temporary recovery in cognition. The alcohol-induced delay was significantly reduced in the high-carbohydrate and high-fat diet groups. Synaptic function exhibited no changes in all groups. Cognitive dysfunction is affected by nutritional status and diet in ALD.

NIK links inflammation to hepatic steatosis by suppressing PPARα in alcoholic liver disease.

Background: Inflammation and steatosis are the main pathological features of alcoholic liver disease (ALD), in which, inflammation is one of the critical drivers for the initiation and development of alcoholic steatosis. NIK, an inflammatory pathway component activated by inflammatory cytokines, was suspected to link inflammation to hepatic steatosis during ALD. However, the underlying pathogenesis is not well-elucidated.Methods: Alcoholic steatosis was induced in mice by chronic-plus-binge ethanol feeding. Both the loss- and gain-of-function experiments by the hepatocyte-specific deletion, pharmacological inhibition and adenoviral transfection of NIK were utilized to elucidate the role of NIK in alcoholic steatosis. Rate of fatty acid oxidation was assessed in vivo and in vitro. PPARα agonists or antagonists of MEK1/2 and ERK1/2 were used to identify the NIK-induced regulation of PPARα, MEK1/2, and ERK1/2. The potential interactions between NIK, MEK1/2, ERK1/2 and PPARα and the phosphorylation of PPARα were clarified by immunoprecipitation, immunoblotting and far-western blotting analysis.Results: Hepatocyte-specific deletion of NIK protected mice from alcoholic steatosis by sustaining hepatic fatty acid oxidation. Moreover, overexpression of NIK contributed to hepatic lipid accumulation with disrupted fatty acid oxidation. The pathological effect of NIK in ALD may be attributed to the suppression of PPARα, the main controller of fatty acid oxidation in the liver, because PPARα agonists reversed NIK-mediated hepatic steatosis and malfunction of fatty acid oxidation. Mechanistically, NIK recruited MEK1/2 and ERK1/2 to form a complex that catalyzed the inhibitory phosphorylation of PPARα. Importantly, pharmacological intervention against NIK significantly attenuated alcoholic steatosis in ethanol-fed mice.Conclusions: NIK targeting PPARα via MEK1/2 and ERK1/2 disrupts hepatic fatty acid oxidation and exhibits high value in ALD therapy.

Alcoholic liver disease: Utility of animal models.

Alcoholic liver disease (ALD) is a major cause of acute and chronic liver injury. Extensive evidence has been accumulated on the pathological process of ALD during the past decades. However, effective treatment options for ALD are very limited due to the lack of suitable in vivo models that recapitulate the full spectrum of ALD. Experimental animal models of ALD, particularly rodents, have been used extensively to mimic human ALD. An ideal animal model should recapitulate all aspects of the ALD process, including significant steatosis, hepatic neutrophil infiltration, and liver injury. A better strategy against ALD depends on clear diagnostic biomarkers, accurate predictor(s) of its progression and new therapeutic approaches to modulate stop or even reverse the disease. Numerous models employing rodent animals have been established in the last decades to investigate the effects of acute and chronic alcohol exposure on the initiation and progression of ALD. Although significant progress has been made in gaining better knowledge on the mechanisms and pathology of ALD, many features of ALD are unknown, and require further investigation, ideally with improved animal models that more effectively mimic human ALD. Although differences in the degree and stages of alcoholic liver injury inevitably exist between animal models and human ALD, the acquisition and translational relevance will be greatly enhanced with the development of new and improved animal models of ALD.

Extracellular vesicles from mice with alcoholic liver disease carry a distinct protein cargo and induce macrophage activation through heat shock protein 90.

Our study indicates a specific protein signature of ALD EVs and demonstrates a functional role of circulating EVs containing heat shock protein 90 in mediating KC/MØ activation in the liver. (Hepatology 2018;67:1986-2000).

Pregnane X receptor promotes ethanol-induced hepatosteatosis in mice

The pregnane X receptor (PXR, NR1I2) is a xenobiotic-sensing nuclear receptor that modulates the metabolic response to drugs and toxic agents. Both PXR activation and deficiency promote hepatic triglyceride accumulation, a hallmark feature of alcoholic liver disease. However, the molecular mechanism of PXR-mediated activation of ethanol (EtOH)-induced steatosis is unclear. Here, using male wildtype (WT) and Pxr-null mice, we examined PXR-mediated regulation of chronic EtOH-induced hepatic lipid accumulation and hepatotoxicity. EtOH ingestion for 8 weeks significantly (1.8-fold) up-regulated Pxr mRNA levels in WT mice. The EtOH exposure also increased mRNAs encoding hepatic constitutive androstane receptor (3-fold) and its target, Cyp2b10 (220-fold), in a PXR-dependent manner. Furthermore, WT mice had higher serum EtOH levels and developed hepatic steatosis characterized by micro- and macrovesicular lipid accumulation. Consistent with the development of steatosis, lipogenic gene induction was significantly increased in WT mice, including sterol regulatory element-binding protein 1c target gene fatty-acid synthase (3.0-fold), early growth response-1 (3.2-fold), and TNFα (3.0-fold), whereas the expression of peroxisome proliferator-activated receptor α target genes was suppressed. Of note, PXR deficiency suppressed these changes and steatosis. Protein levels, but not mRNAs levels, of EtOH-metabolizing enzymes, including alcohol dehydrogenase 1, aldehyde dehydrogenase 1A1, and catalase, as well as the microsomal triglyceride transfer protein, involved in regulating lipid output were higher in Pxr-null than in WT mice. These findings establish that PXR signaling contributes to ALD development and suggest that PXR antagonists may provide a new approach for ALD therapy.

Nrf2 Activation Is Required for Ligustrazine to Inhibit Hepatic Steatosis in Alcohol-Preferring Mice and Hepatocytes.

Hepatic steatosis is the most distinctive feature of alcoholic liver disease (ALD). Our previous in vivo study showed that ligustrazine, a major active alkaloid isolated from Ligusticum chuanxiong Hort, attenuated alcohol-induced hepatic steatosis and in vitro study revealed that nuclear factor (erythroid-derived 2)-like 2 (Nrf2) activation might be a prerequisite. This study was aimed to explore the in vivo function of Nrf2 in the protective effect of ligustrazine and illustrate downstream mechanisms. Nrf2 shRNA lentivirus was introduced to establish Nrf2-knockdown mice. Results showed that Nrf2 knockdown aggravated alcoholic liver injury and abolished the protective effect of ligustrazine, evidenced by elevated serum biomarkers and severe liver inflammation. Ligustrazine impressively ameliorated hepatic steatosis through inhibiting hepatic sterol regulatory element-binding protein-1c and inducing peroxisome proliferator-activated receptor-alpha, which was abrogated by Nrf2 knockdown. Noteworthily, Nrf2 knockdown apparently reinforced the inductive effect of alcohol on hypoxia-inducible factor 1-alpha (HIF-1α). Ligustrazine weakened the stimulation of alcohol on HIF-1α expression, which was abrogated by Nrf2 shRNA lentivirus. Consistent results were obtained from in vitro study, implying that Nrf2/HIF-1α pathway participated in the modulation of ligustrazine. Gain- or loss-of-function analyses further revealed that Nrf2 siRNA and dimethyloxalylglycine, a HIF-1α agonist, abolished the inhibitory effect of ligustrazine, whereas HIF-1α siRNA mimicked the role of ligustrazine and even rescued the inhibitory effect of ligustrazine on ethanol-induced lipid accumulation in Nrf2-knockdown hepatocytes. Taken together, we concluded that ligustrazine attenuated alcohol-induced hepatic steatosis via a Nrf2/HIF-1α pathway-dependent mechanism.

Metabolic Features of Alcoholic Liver Disease.

Alcoholic Liver Disease (ALD) can be defined as the hepatic manifestations caused by excessive alcohol intake. ALD comprises a spectrum from simple steatosis to cirrhosis. To review diverse clinical and metabolic characteristics and their impact in ALD. The concurrence of metabolic alterations, including obesity and diabetes, and alcohol consumption increase the risk of liver injury and also the morbidity and mortality associated with chronic liver disease.

Therapeutic Benefits of Spleen Tyrosine Kinase Inhibitor Administration on Binge Drinking-Induced Alcoholic Liver Injury, Steatosis, and Inflammation in Mice.

Binge drinking is increasingly recognized as an important cause of liver disease with limited therapeutic options for patients. Binge alcohol use, similar to chronic alcohol consumption, induces numerous deregulated signaling events that drive liver damage, steatosis, and inflammation. In this article, we evaluated the role of spleen tyrosine kinase (SYK), which modulates numerous signaling events previously identified linked in the development alcohol-induced liver pathology. A 3-day alcohol binge was administered to C57BL/6 female mice, and features of alcoholic liver disease were assessed. Some mice were treated daily with intraperitoneal injections of a SYK inhibitor (R406; 5 to 10mg/kg body weight) or drug vehicle control. Liver and serum samples were collected and were assessed by Western blotting, biochemical, ELISA, electrophoretic mobility shift assays, real-time quantitative polymerase chain reaction, and histopathological analysis. We found that binge drinking induced significant SYK activation (SYK(Y525/526) ) with no change in total SYK expression in the liver. Functional inhibition of SYK activation using a potent SYK inhibitor, R406, was associated with a significant decrease in alcohol-induced hepatic inflammation as demonstrated by decreased phospho-nuclear factor kappa beta (NF-κB) p65, NF-κB nuclear binding, tumor necrosis factor-alpha, and monocyte chemoattractant protein-1 mRNA in the liver. Compared to vehicle controls, SYK inhibitor treatment decreased alcohol binge-induced hepatocyte injury indicated by histology and serum alanine aminotransferase. Strikingly, SYK inhibitor treatment also resulted in a significant reduction in alcohol-induced liver steatosis. Our novel observations demonstrate the role of SYK, activation in the pathomechanism of binge drinking-induced liver disease highlighting SYK a potential multifaceted therapeutic target.

Tetramethylpyrazine prevents ethanol-induced hepatocyte injury via activation of nuclear factor erythroid 2-related factor 2.

Inhibiting the major features of alcoholic liver disease (ALD) such as lipid accumulation and oxidative stress is a promising strategy of treating ALD. Tetramethylpyrazine (TMP) has curative effects on various diseases. However, the effects of TMP on ethanol-induced hepatocyte injury and the related mechanisms remain unclear. The aim of this study is to elucidate the effects of TMP and the potential mechanisms in vitro. Ethanol-stimulated LO2 cells were used as an in vitro model of ALD. Several biomarkers related to cell injury, lipid accumulation, and oxidative stress were determined to evaluate the effects of TMP. Nuclear factor erythroid 2-related factor 2 (Nrf2) expression plasmids and Nrf2 small interfering RNA (siRNA) were used to establish the role of Nrf2. TMP increased Nrf2 expression and nuclear translocation. TMP prevented ethanol-induced hepatocyte injury, as indicated by the enhanced cell viability, reduced activities of aspartate transaminase and alanine aminotransferase in the culture medium, and inhibition of cell apoptosis. Furthermore, TMP reduced the levels of lipid droplets, triglyceride, and total cholesterol, probably by regulating genes related to lipid metabolism. Besides, TMP alleviated ethanol-induced oxidative stress by increasing superoxide dismutase activity and the glutathione level and by reducing the levels of reactive oxygen species and malondialdehyde. In addition, overexpression of Nrf2 enhanced the effects of TMP on cell injury, lipid accumulation, and oxidative stress, whereas Nrf2 siRNA eliminated the effects of TMP. TMP prevents ethanol-induced hepatocyte injury by inhibiting lipid accumulation and oxidative stress, and via an Nrf2 activation-dependent mechanism.

PTH-094 Are megamitochondria a cellular survival strategy for ethanol-induced liver toxicity?

Introduction A characteristic histological feature of alcoholic liver disease (ALD), known since the 1970s, is the presence of megamitochondria (MM) in hepatocytes. In a recent study conducted in more than 100 patients, the presence of MM was associated with a lower risk of death within 90 days.1Mitochondria are dynamic organelles and their shape is intimately involved with their function, under physiological conditions the changes in mitochondrial morphology are regulated by mitochondrial-shaping proteins (MSP). However, the mechanisms driving MM formation, their role, function and involvement in the pathogenesis of ALD is currently unknown, which is the aim of this study. Method Human precision cut liver slices (PCLS) and hepatoma cells VL-17A (positive for ADH/CYP2E1) were cultured with EtOH alone or in combination with FFA, to induce hepatotoxicity. Cell viability was evaluated by FACS (AnnexinV/PI positive cells) or ATP quantification. Post-treatment, changes in the mitochondrial shape were assessed by confocal and electron microscopy; MSP expression and activation was also analysed by Western blot on isolated mitochondria. Additionally, mitochondrial functionality was evaluated in intact cells using the Seahorse Bioscience analyser. Changes in inflammatory cytokine production were assessed by Cytokine Bead Array. Results The toxic effect of EtOH (+/-FFA) was confirmed by an increase in the percentage of dead cells or a reduction in ATP levels. Cells that survived the toxic insult showed a dramatically increased proportion of MM; however they also had a compromised ability to grow. Mitochondrial functionality, in terms of coupling efficiency and spare respiratory capacity, was also impaired. Pro inflammatory cytokine profile was found in the supernatants of these cultures. This was associated with a reduced activity in the MSP related to mitochondrial fragmentation. Conclusion Cellular survival strategies, in the face of a chronic toxic stimulus, are a necessary requirement to ensure continuing functionality of the organ. We show here that growth retarded hepatocytes, which survived the EtOH insult, are characterised by a dramatic formation of MM, suggesting this as a mechanism to avoid death. We also show for the first time the involvement of MSP in this pathway, raising the potentiality for the use of these proteins as therapeutic targets and biomarkers in ALD. Disclosure of interest None Declared. ![Abstract PTH-094 Figure 1][1] Abstract PTH-094 Figure 1 Ethanol-induced megamitochondria (MM) formation visualised (a) by electron microscopy in human PCLS and (b) by confocal microscopy in VL-17A cells Reference 1. Altamirano, et al . Gastroenterology , 2014;146(5):1231–1239 [1]: pending:yes

Pathology of Alcoholic Liver Disease.

Alcohol-attributable burden on global health is increasing, and the relationship between population alcohol consumption and liver-related deaths is strong. Longstanding scientific and clinical work has led to a relatively thorough, if not complete, understanding of the effects of alcohol consumption on the liver. Pathologic features of alcoholic liver disease (ALD) are recognized by pathologists and used to assist clinicians in diagnosing and determining severity of disease in patients suspected of ALD. In this review, we discuss the pathologic manifestations of ALD and provide salient points on their pathophysiology. In addition, the benefits and indications of liver biopsy and important differential diagnoses, including features distinguishing these entities, are reviewed.

A Public Health Issue that Increased Prevelance: Non-Acholic Fatty Liver Disease

Non-alcoholic fatty liver disease (NAFLD), is a liver disease, that occur in non alcohol users, with same histological features of alcoholic liver disease. The increased importance of NAFLD is depends on its close relation of current problems such as Type 2 Diabetes Mellitus and hyperlipidemia, and its high prevalence as %20-30, in Europe and Middle East. NAFLD has a wide spectrum from simple steatozis (SS), liver cell damage, non alcoholic steatohepatitis (NASH) with inflammation to high stage fibrosis, and cirrhosis. Hepathocellular cancer and liver failure might develop in cirrhosis patients. Biopsy is gold standard for NAFLD diagnosis; differentiate from SS and NASH and defining NASH stages. Nutrition regulations and slow and continuous weight loss with regular exercises is still best treatment method

Monocytosis Correlated With Acute Alcoholic Hepatitis: A Case Report and Literature Review

Introduction Alcohol is a most frequent cause of liver disease in western countries. Alcoholic hepatitis is observed in approximately 20% of heavy drinkers. Acute alcoholic hepatitis (AH) is associated with mortality as high as 50%.Up to 40% of patient with severe alcoholic hepatitis die within 6 months after the onset of the clinical syndrome. Identifying individuals with a high mortality risk is crucial in the management of acute alcoholic hepatitis. We observed a patients with acute alcoholic hepatitis tend to have an increased level of monocytes. Case presentation 47-year-old male with a significant history of alcohol abuse for 30 years presented with confusion, generalized shaking, and tremors for 4 days was admitted to the hospital. The patient was hospitalized 6 months ago for acute alcoholic hepatitis. Physical examination: Bp 140/80mmHg, Ultrasound of Abdomen showed hepatomegaly with enlarged liver measuring 19.1 cm in the midclavicular line and hepatic steatosis and gallbladder sludge without evidence of wall thickening or pericystic fluid. CT of abdomen Suspected fatty infiltration of the liver. Patient had ammonia level of 56 -58. His MELD score was between 52 in which the alcoholic hepatitis was diagnosed. The patient was treated with Ativan, Chlordiazepoxide, Thiamine, folic acid, Mulativitamin, and lactulose. After patient was treated for 7 days, Symptoms had been well controlled, and all of tests went to back normal range. Table1 shows clinical data of current and previous admission. Discussion Alcoholics develop acute hepatitis as an inflammatory reaction to the cells affected by fatty change. Diagnosis of alcoholic hepatitis based on clinical symptoms and laboratory finding alone including elevated AST/ALT (but may &lt; 300 IU/mL), AST&gt;ALT of 2, Total serum bilirubin &gt; 5mg/deciliter, elevated INR, thrombocytopenia, and hypoalbuminimia, (cirrhosis). Leukocytosis with neutophilic predominance has been reported to correlates with degree of injury. As a feature of alcoholic liver disease, monocytosis was first reported on 1983, however, the mechanism mediates this clinical feature has been unclear. Recently data has been shown that activated monocytes have been postulated to play an important role in the pathogenesis of alcoholic liver disease (ALD), in which can produce Interleukin-1 (IL-1) induces interleukin-6 (IL-6) production during acute alcoholic liver injury phase reactant. The number of monocytes, one of the most important components of the inflammatory process in ALD maybe as an independent marker that can be utilized to determine the disease severity and predict outcome of the patients. Conclusion Our result suggests that monocytosis is associated with acute alcoholic hepatitis (Table 1). Further research may provide us with a better understanding of the clinical scenario and help elucidate the best prevention and treatment options of alcohol–related liver disease. Disclosures: No relevant conflicts of interest to declare.

Globular adiponectin inhibits ethanol-induced apoptosis in HepG2 cells through heme oxygenase-1 induction

Hepatocellular apoptosis is an essential pathological feature of alcoholic liver disease. Adiponectin, an adipokine predominantly secreted from adipose tissue, has been shown to play beneficial roles in alcoholic liver disease against various inflammatory and pro-apoptotic molecules. However, the effects of adiponectin on ethanol-induced apoptosis in liver cells are largely unknown. Herein, we investigated the role of globular adiponectin (gAcrp) in the prevention of ethanol-induced apoptosis and further tried to decipher the potential mechanisms involved. In the present study, we demonstrated that gAcrp significantly inhibits both ethanol-induced increase in Fas ligand expression and activation of caspase-3 in human hepatoma cell lines (HepG2 cells), suggesting that gAcrp plays a protective role against ethanol-induced apoptosis in liver cells. This protective effect of gAcrp was mediated through adiponectin receptor R1 (adipoR1). Further, globular adiponectin treatment caused induction of heme oxygenase-1 (HO-1) through, at least in part, nuclear factor (erythroid-derived 2)-like 2, (Nrf2) signaling. Treatment with SnPP, a pharmacological inhibitor of HO-1, and knockdown of HO-1 with small interfering RNA (siRNA) restored caspase-3 activity suppressed by gAcrp, indicating a critical role of HO-1 in mediating the protective role of gAcrp in ethanol-induced apoptosis in liver cells. In addition, carbon monoxide, a byproduct obtained from the catabolism of free heme was found to contribute to the anti-apoptotic effect of adiponectin. In conclusion, these data demonstrated that globular adiponectin prevents ethanol-induced apoptosis in HepG2 cells via HO-1 induction and revealed a novel biological response of globular adiponectin in the protection of liver injury from alcohol consumption.

Autophagy Reduces Acute Ethanol-Induced Hepatotoxicity and Steatosis in Mice

Alcohol abuse is a major cause of liver injury. The pathologic features of alcoholic liver disease develop over prolonged periods, yet the cellular defense mechanisms against the detrimental effects of alcohol are not well understood. We investigated whether macroautophagy, an evolutionarily conserved cellular mechanism that is commonly activated in response to stress, could protect liver cells from ethanol toxicity. Mice were acutely given ethanol by gavage. The effects of ethanol on primary hepatocytes and hepatic cell lines were also studied in vitro. Ethanol-induced macroautophagy in the livers of mice and cultured cells required ethanol metabolism, generation of reactive oxygen species, and inhibition of mammalian target of rapamycin signaling. Suppression of macroautophagy with pharmacologic agents or small interfering RNAs significantly increased hepatocyte apoptosis and liver injury; macroautophagy therefore protected cells from the toxic effects of ethanol. Macroautophagy induced by ethanol seemed to be selective for damaged mitochondria and accumulated lipid droplets, but not long-lived proteins, which could account for its protective effects. Increasing macroautophagy pharmacologically reduced hepatotoxicity and steatosis associated with acute ethanol exposure. Macroautophagy protects against ethanol-induced toxicity in livers of mice. Reagents that modify macroautophagy might be developed as therapeutics for patients with alcoholic liver disease.

Hepatic macrophage iron aggravates experimental alcoholic steatohepatitis

One prime feature of alcoholic liver disease (ALD) is iron accumulation in hepatic macrophages/Kupffer cells (KC) associated with enhanced NF-kappaB activation. Our recent work demonstrates a peroxynitrite-mediated transient rise in intracellular labile iron (ILI) as novel signaling for endotoxin-induced IKK and NF-kappaB activation in rodent KC. The present study investigated the mechanism of KC iron accumulation and its effects on ILI response in experimental ALD. We also tested ILI response in human blood monocytes. Chronic alcohol feeding in rats results in increased expression of transferrin (Tf) receptor-1 and hemochromatosis gene (HFE), enhanced iron uptake, an increase in nonheme iron content, and accentuated ILI response for NF-kappaB activation in KC. Ex vivo treatment of these KC with an iron chelator abrogates the increment of iron content, ILI response, and NF-kappaB activation. The ILI response is evident in macrophages derived from human blood monocytes by PMA treatment but not in vehicle-treated monocytes, and this differentiation-associated phenomenon is essential for maximal TNF-alpha release. PMA-induced macrophages load iron dextran and enhance ILI response and TNF-alpha release. These effects are reproduced in KC selectively loaded in vivo with iron dextran in mice and more importantly aggravate experimental ALD. Our results suggest enhanced iron uptake as a mechanism of KC iron loading in ALD and demonstrate the ILI response as a function acquired by differentiated macrophages in humans and as a priming mechanism for ALD.