Alcoholic Liver Steatosis Research Articles

N-3 Polyunsaturated Fatty Acids Protect against Alcoholic Liver Steatosis by Activating FFA4 in Kupffer Cells.

Supplementation with fish oil rich in omega-3 polyunsaturated fatty acids (n-3 PUFAs) effectively reduces acute and chronic alcohol-induced hepatic steatosis. We aimed to find molecular mechanisms underlying the effects of n-3 PUFAs in alcohol-induced hepatic steatosis. Because free fatty acid receptor 4 (FFA4, also known as GPR120) has been found as a receptor for n-3 PUFAs in an ethanol-induced liver steatosis model, we investigated whether n-3 PUFAs protect against liver steatosis via FFA4 using AH7614, an FFA4 antagonist, and Ffa4 knockout (KO) mice. N-3 PUFAs and compound A (CpdA), a selective FFA4 agonist, reduced the ethanol-induced increase in lipid accumulation in hepatocytes, triglyceride content, and serum ALT levels, which were not observed in Ffa4 KO mice. N-3 PUFAs and CpdA also reduced the ethanol-induced increase in lipogenic sterol regulatory element-binding protein-1c expression in an FFA4-dependent manner. In Kupffer cells, treatment with n-3 PUFA and CpdA reversed the ethanol-induced increase in tumor necrosis factor-α, cyclooxygenase-2, and NLR family pyrin domain-containing 3 expression levels in an FFA4-dependent manner. In summary, n-3 PUFAs protect against ethanol-induced hepatic steatosis via the anti-inflammatory actions of FFA4 on Kupffer cells. Our findings suggest FFA4 as a therapeutic target for alcoholic hepatic steatosis.

Molecular insights on intracellular Wnt/β-catenin signaling in alcoholic liver disease.

Alcoholic liver disease (ALD) is one of the most common health problems worldwide, especially in developing countries caused by chronic consumption of alcohol on a daily basis. The ALD spectrum is initiated with the early stages of alcoholic fatty liver (steatosis), progressing to alcoholic steatohepatitis, followed by the later stages of fibrosis and in some cases, cirrhosis and hepatocellular carcinoma (HCC). The Wnt/β-catenin signaling required for healthy liver development, function, and regeneration is found to be aberrated in ALD, attributed to its progression. This review is to elucidate the association of Wnt/β-catenin signaling with various stages of ALD progression. Alcohol causes downregulation of Wnt/β-catenin signaling components and thereby suppressing the pathway. Reports have been published that aberrated Wnt/β-catenin signaling, especially the absence of β-catenin, results in decreased alcohol metabolism, causing steatosis followed by steatohepatitis via lipid accumulation, lipid peroxidation, liver injury, increased oxidative stress and apoptosis of hepatocytes, contributing to the advancement of ALD. Contrastingly, the progression of later stages of ALD like fibrosis and HCC depends on the increased activation of Wnt/β-catenin signaling and its components. Existing studies reveal the varied expression of Wnt/β-catenin signaling in ALD. However, the dual role of the Wnt/β-catenin pathway in earlier and later stages of ALD is not clear. Therefore, studies on the Wnt/β-catenin pathway and its components in various manifestations of ALD might provide insight in targeting the Wnt/β-catenin pathway in ALD treatment.

A review: Alcoholic liver disease pathophysiology, current molecular and clinical perspectives

Alcohol-related liver damage (ALD) is a term used for a variety of liver conditions comprising cirrhosis, fibrosis, and steatohepatitis with chronic inflammation that is predominantly brought on by heavy alcohol consumption. ALD is currently regarded as one of the most common causes of liver disease-related mortality on a global scale. The majority of cirrhosis in the liver cases seen in district general hospitals in the UK are caused by alcohol, which is a key factor in the Western world. Alcoholic cirrhosis, acute alcoholic hepatitis, and alcoholic fatty liver (steatosis) are the three most widely recognized types of alcoholic liver disease. According to the National Institute on Alcohol Abuse and Alcoholism's the most recent surveillance report, liver cirrhosis was among the top twelve causes of death in the country, contributing to a total of 14,477 deaths in the year 2014.This page speaks about the various types of alcoholic liver damage (ALD), with a focus on cirrhosis because it is the type of liver diseases that is most frequently linked to alcohol misuse and has the most studies investigating it. Epidemiological studies have assessed the incidence of ALD and the factors who frequently cause the condition. Although excessive alcohol use is the main cause of ALD, gender and cultural variations also have a significant role in the prevalence of liver disease. Since the 1970s, cirrhosis-related mortality rates have decreased in the United States and certain other countries as well. This dip may have been caused by a variety of variables, including increasing enrollment in alcohol rehab facilities and in Alcoholics Anonymous, drops in alcohol consumption, and more

Hesperitin attenuates alcoholic steatohepatitis by regulating TLR4/NF-κB signaling in mice

In the peel of citrus (Rutaceae) fruit, hesperitin (Hesp), a flavanone glycoside chemical, is found naturally. Hesp has been found to have a wide range of pharmacological actions, including anti-inflammatory, antioxidant, antiviral, and anticancer properties, according to earlier research. However, nothing is known regarding its function in alcoholic liver steatosis and inflammation. In this study, we employed a network pharmacology approach to identify the TLR4 signaling pathway as a primary target of Hesp for the treatment of alcoholic steatohepatitis (ASH). Molecular docking results showed that Hesp bound to the representative target TLR4 and exhibited good affinity. In addition, Hesp inhibits the TLR4 target and consequently the NF-κB signaling pathway, which in turn slows the evolution of alcoholic steatohepatitis, according to further in vitro and in vivo tests. The results of this study preliminarily indicate that Hesp is an ideal drug candidate for the treatment of ASH.

Inhibition of ACSS2 attenuates alcoholic liver steatosis via epigenetically regulating de novo lipogenesis.

Steatosis is the early pathological change in alcohol-associated liver disease. However, its precise mechanism is still unclear. The present study is aimed to explore the role and mechanism of acetyl-CoA synthetase 2 (ACSS2) in acute alcohol-induced lipogenesis. The increase in ACSS2 nuclear import and histone H3 acetylation were observed in mice after intraperitoneally injected with 2 g/kg ethanol or oral administration of 5 g/kg ethanol and also validated in hepatocytes stimulated with ethanol or acetate. The role of ACSS2 was further explored in liver-specific ACSS2 knockdown mice fed with ethanol-containing diet. Alcohol challenge induced hepatic lipid deposition and upregulated lipogenic genes in mice. It also promoted ACSS2 nuclear import and increased histone H3 acetylation. In hepatocytes, ethanol induced similar phenomena whereas ACSS2 knockdown blocked histone acetylation and lipogenic gene induction. P300/CBP associated factor (PCAF), but not general control nonderepressible 5, CREB-binding protein (CBP) and p300, facilitated H3K9 acetylation responding to ethanol challenge. CUT&RUN assay showed the enrichment of acetylated histone H3K9 surrounding Fasn and Acaca promoters. These results indicated that ethanol metabolism promoted ACSS2 nuclear import to support lipogenesis via H3K9 acetylation. In alcohol-feeding mice, liver-specific ACSS2 knockdown blocked the interaction between PCAF and H3K9 and suppressed lipogenic gene induction in the liver, demonstrating the critical role of ACSS2 in lipogenesis. Our study demonstrated that alcohol metabolism generated acetyl-CoA in the nucleus dependently on nuclear ACSS2, contributing to epigenetic regulation of lipogenesis in hepatic steatosis. Targeting ACSS2 may be a potential therapeutical strategy for acute alcoholic liver steatosis.

A clinical experience-based Chinese herbal formula improves ethanol-induced drunken behavior and hepatic steatohepatitis in mice models

BackgroundBao-Gan-Xing-Jiu-Wan (BGXJW) is a clinical experience-based Chinese herbal formula. Its efficacy, pharmacological safety, targeted function, process quality, and other aspects have met the evaluation standards and the latest requirements of preparations. It could prevent and alleviate the symptoms of drunkenness and alcoholic liver injury clinically. The present work aims to elucidate whether BGXJW could protect against drunkenness and alcoholic liver disease in mice and explore the associated mechanism.Material and methodsWe used acute-on-chronic (NIAAA) mice model to induce alcoholic steatosis, and alcohol binge-drinking model to reappear the drunk condition. BGXJW at indicated doses were administered by oral gavage respectively to analyze its effects on alcoholic liver injury and the associated molecular mechanisms.ResultsBGXJW had no cardiac, hepatic, renal, or intestinal toxicity in mice. Alcoholic liver injury and steatosis in the NIAAA mode were effectively prevented by BGXJW treatment. BGXJW increased the expression of alcohol metabolizing enzymes ADH, CYP2E1, and ALDH2 to enhance alcohol metabolism, inhibited steatosis through regulating lipid metabolism, counteracted alcohol-induced upregulation of lipid synthesis related proteins SREBP1, FASN, and SCD1, meanwhile it enhanced fatty acids β-oxidation related proteins PPAR-α and CPT1A. Alcohol taken enhanced pro-inflammatory TNF-α, IL-6 and down-regulated the anti-inflammatory IL-10 expression in the liver, which were also reversed by BGXJW administration. Moreover, BGXJW significantly decreased the blood ethanol concentration and alleviated drunkenness in the alcohol binge-drinking mice model.ConclusionsBGXJW could effectively relieve drunkenness and prevent alcoholic liver disease by regulating lipid metabolism, inflammatory response, and alcohol metabolism.Graphical

PREVALENCE OF COMBINED LIVER DAMAGE IN MEXICAN POPULATION

Metabolic syndrome and alcohol consumption are the two main causes of liver steatosis. Often, one of them predominates for its development and the other acts as a cofactor. The impact of these entities separately and their damage in synergy in the Mexican population is currently unknown. Our objective is to determine the prevalence of non-alcoholic liver steatosis, alcoholic liver steatosis, and dual damage in donors from the blood bank of the Hospital General de México “Dr. Eduardo Liceaga” (HGMEL) using transient elastography (TE). Pilot study. prospective, transversal, descriptive, and analytical. Healthy donors from the HGMEL blood bank who attended from June 8 to 29 2021 ≥18 years old, of any gender, will be included with a body mass index (BMI) ≥18.5. Donors with liver disease of any etiology and liver malignancy will be excluded. Donors who did not attend TE will be eliminated. The equipment used for TE was FibroScan® 502 Touch. Descriptive statistics will be used with measures of central tendency and dispersion. 30 donors were recruited, four were eliminated for not having performed TE. The age of the subjects was 36.53 ± 12.13 years. There were 13 female subjects and 13 males. Seven (26.92%) donors were classified with non-alcoholic fatty liver disease (NAFLD), 2 (7.69%) with alcoholic fatty liver disease (ALD), and 1 (3.84%) subject with liver steatosis due to combined damage; the 16 (61.55%) remaining subjects corresponded to a healthy population. From the NAFLD group, S1 steatosis was documented in 1 subject (14.28%), S2 in 2 (28.56%), and S3 in 4 (57.16%); F0-F1 liver stiffness was found in 6 (85.72%) subjects, and F4 in 1 (14.28%); Of this group, 3 (42.86%) subjects were classified as overweight, 3 (42.86%) with grade 2 obesity, and 1 (14.28%) with grade 3 obesity. On the other hand, from the ALD group, one donor was found with steatosis S1, and 1 with S2; one of them had liver stiffness F0-F1 with an AUDIT score of 17 and a risk pattern for excessive alcohol consumption (117 g 2 to 4 times a month), while the other subject had F4 fibrosis with an AUDIT score of 7 and a pattern risk of excessive consumption (208 g 2 to 4 times per month), both consume beer. Finally, one donor had liver steatosis due to combined damage, with S3 for steatosis and F4 liver stiffness (27.4 kPa, the highest record in the study); He has grade 3 obesity (BMI 44.41), an AUDIT score of 4, and a pattern of excessive consumption (78 g 2 to 4 times a month) of distillates. What is relevant about this work is that it studies an apparently healthy population and, according to our results, 38.45% (10) of the subjects already have a degree of liver steatosis and even 11.53% (3) showed advanced fibrosis. Our findings are similar to others about the prevalence of nonalcoholic liver steatosis, such as the work of the López-Velázquez group (26%) in 2014. Our study is novel in the investigation of alcoholic liver steatosis and the combined damage. Those issues are scarce addressed in our country. We also collect additional information, such as the identification of the consumption pattern of people with risky alcohol intake and the type of drink they consume. Our study shows that liver steatosis in this sample of the Mexican population is predominantly non-alcoholic; all subjects within this group have high BMI. Excessive risk consumption is prevalent in subjects with alcohol-related steatosis, and beer is the most frequent drink. In combined damage, obesity and excessive consumption of distillates seem to be the cause of severe steatosis. The only patient in this group already shows advanced fibrosis, which makes us suspect that dual damage accelerates the rate of fibrosis progression. The authors declare that there is no conflict of interest.

CYP2E1 in Alcoholic and Non-Alcoholic Liver Injury. Roles of ROS, Reactive Intermediates and Lipid Overload.

CYP2E1 is one of the fifty-seven cytochrome P450 genes in the human genome and is highly conserved. CYP2E1 is a unique P450 enzyme because its heme iron is constitutively in the high spin state, allowing direct reduction of, e.g., dioxygen, causing the formation of a variety of reactive oxygen species and reduction of xenobiotics to toxic products. The CYP2E1 enzyme has been the focus of scientific interest due to (i) its important endogenous function in liver homeostasis, (ii) its ability to activate procarcinogens and to convert certain drugs, e.g., paracetamol and anesthetics, to cytotoxic end products, (iii) its unique ability to effectively reduce dioxygen to radical species causing liver injury, (iv) its capability to reduce compounds, often generating radical intermediates of direct toxic or indirect immunotoxic properties and (v) its contribution to the development of alcoholic liver disease, steatosis and NASH. In this overview, we present the discovery of the enzyme and studies in humans, 3D liver systems and genetically modified mice to disclose its function and clinical relevance. Induction of the CYP2E1 enzyme either by alcohol or high-fat diet leads to increased severity of liver pathology and likelihood to develop ALD and NASH, with subsequent influence on the occurrence of hepatocellular cancer. Thus, fat-dependent induction of the enzyme might provide a link between steatosis and fibrosis in the liver. We conclude that CYP2E1 has many important physiological functions and is a key enzyme for hepatic carcinogenesis, drug toxicity and liver disease.

Livogrit ameliorates acetaldehyde-induced steatosis in HepG2 cells through modulation of lipogenesis and β-oxidation pathways

BackgroundFrequent alcohol consumption leads to development of alcoholic liver steatosis. Onset of steatosis is caused by alcohol metabolite ‘acetaldehyde’ causing intracellular accumulation of lipid bodies due to abnormal metabolism. Livogrit is a tri-herbal Ayurveda formulation prepared using Boerhavia diffusa L., Phyllanthus niruri L., and Solanum nigrum L. plant extracts. The formulation has earlier proven effective against carbon tetrachloride-induced toxicity and non-alcoholic fatty acid induced liver steatosis. PurposeIn present study, we evaluated efficacy of Livogrit in ameliorating ‘acetaldehyde’ induced steatosis and assessed its mode of action. MethodHepG2 cells were exposed to varying concentrations of acetaldehyde and assessed for lipid body accumulation and cytosafety. An inhibition concentration of 50% for acetaldehyde was observed at 71.42 mM in HepG2 cells. 1 mM acetaldehyde was selected as a non-cytotoxic steatosis-stimulating dose. The acetaldehyde-stimulated HepG2 cells were used for studying the disease-reversing efficacy of Livogrit. Efficacy of the formulation was analyzed at cellular, biochemical, and molecular levels. ResultsAcetaldehyde stimulation caused an increase in intracellular lipid and triglyceride accumulation, release of ALT and AST enzymes, ROS generation, and decrease in extracellular release of glycerol molecules in HepG2 cells. Livogrit treatment of the stimulated-HepG2 cells showed a reversal of steatosis through normalization of observed intra- and extracellular parameters. At gene level, Livogrit-modulated the acetaldehyde-stimulated upregulation of SREBP1c, FAS, PLIN2, TNF-α, NF-kB, and LC3A genes associated with lipogenesis, inflammation, and autophagy. At protein level, the protective role of Livogrit was observed in acetaldehyde-stimulated HepG2 cells through modulation of TNF-α, PLIN2, and LC3 proteins. Livogrit provided further protection against steatosis through stimulation of fatty acid β-oxidation associated with SIRT1, CPT1A, and PPAR-α genes. ConclusionIn conclusion, Livogrit recovered HepG2 cells from alcoholic steatosis at biochemical and molecular levels through multifaceted modulation of lipogenesis, inflammation, autophagy, and β-oxidation. Hence, Livogrit was found to be a potential candidate for the amelioration of alcohol-induced steatosis.

Roles of Nrf2 in Liver Diseases: Molecular, Pharmacological, and Epigenetic Aspects

Liver diseases represent a critical health problem with 2 million deaths worldwide per year, mainly due to cirrhosis and its complications. Oxidative stress plays an important role in the development of liver diseases. In order to maintain an adequate homeostasis, there must be a balance between free radicals and antioxidant mediators. Nuclear factor erythroid 2-related factor (Nrf2) and its negative regulator Kelch-like ECH-associated protein 1 (Keap1) comprise a defense mechanism against oxidative stress damage, and growing evidence considers this signaling pathway as a key pharmacological target for the treatment of liver diseases. In this review, we provide detailed and updated evidence regarding Nrf2 and its involvement in the development of the main liver diseases such as alcoholic liver damage, viral hepatitis, steatosis, steatohepatitis, cholestatic damage, and liver cancer. The molecular and cellular mechanisms of Nrf2 cellular signaling are elaborated, along with key and relevant antioxidant drugs, and mechanisms on how Keap1/Nrf2 modulation can positively affect the therapeutic response are described. Finally, exciting recent findings about epigenetic modifications and their link with regulation of Keap1/Nrf2 signaling are outlined.

Noninvasive 40-Hz Light Flicker Rescues Circadian Behavior and Abnormal Lipid Metabolism Induced by Acute Ethanol Exposure via Improving SIRT1 and the Circadian Clock in the Liver-Brain Axis.

Sirtuin 1 (SIRT1) is a protein deacetylase with important cellular functions, as it regulates numerous processes, including the circadian rhythm in peripheral tissues. Efforts are ongoing to reveal how Sirt1 can be used to treat diseases, such as alcoholic liver disease (ALD), Alzheimer's disease, and liver fibrosis. We have recently shown that noninvasive exposure to 40-Hz light flicker activates hypothalamic SIRT1 gene expression, thereby regulating the central circadian clock. This study investigated the effects of 40-Hz light flicker in a mouse model of ALD. RNA sequencing (RNA-seq) analysis was performed to explore the potential pathways affected by 40-Hz light flicker. We found that 40-Hz light flicker significantly decreased the acute ethanol-induced increases in serum alanine aminotransferase (ALT) and serum triglyceride (TG) levels and reduced fat-droplet accumulation in mouse livers. Additionally, 40-Hz light flicker significantly suppressed ethanol-induced increases in sterol regulatory element binding protein 1 (SREBP-1) and fatty acid synthase (Fasn) levels. Furthermore, the ethanol induced significant decreases in both Sirt1 levels and phosphorylation of adenosine monophosphate-activated protein kinase subunit (AMPKα), compared with those in the control group. Strikingly, pretreatment with 40-Hz light flicker ameliorated such ethanol-induced decreases in SIRT1 levels and AMPKα phosphorylation. In addition, ethanol-induced increases in levels of brain and muscle arnt-like protein-1 (BMAL1), circadian locomotor output cycles kaput (CLOCK), and period 2 (PER2) were reversed by 40-Hz light flicker. RNA-seq analysis revealed significant differences in expression of genes related to the AMPK signalling. Moreover, ethanol consumption altered mRNA levels of Sirt1 and circadian genes in the suprachiasmatic nucleus (SCN), indicating that ethanol influenced central pacemaker genes; however, 40-Hz light flicker reversed these ethanol-induced changes. Taken together, our findings demonstrate that 40-Hz light flicker rapidly influence the SCN and exhibits inhibitory properties on hepatic lipogenesis, indicating that 40-Hz light flicker has therapeutic potential for preventing alcoholic liver steatosis.

Regulation of CD39 expression in ATP-P2Y2R-mediated alcoholic liver steatosis and inflammation

Inflammation plays a central role in the progression of alcoholic liver disease. ATP-P2Y2R signaling and CD39 play an important role in various diseases, but little is known about their role in alcoholic liver steatosis and inflammation. As a transmembrane hydrolase, CD39 hydrolyzes ATP, while the mutual regulation of CD39 and ATP-P2Y2R in alcoholic steatohepatitis is poorly understood. Here, we found that the expression of ATP, P2Y2R, and CD39 is increased significantly both in the liver of alcohol-fed mice and alcohol-induced RAW264.7 cell lines. In this study, C57BL/6 mice were intrapretationally injected with P2Y2R inhibitor suramin from day 4 until day 10 during the induction of a chronic/binge drinking model. Pharmacological blockade of P2Y2R largely prevents liver damage, lipid accumulation, and inflammation, with concomitant down-expression of CD39 in liver. We found that the inhibition of P2Y2R in vitro reduces inflammation via down-expression of interleukin 6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-alpha (TNF-α), and the expression of CD39 was reduced, whereas the activation of P2Y2R showed an opposite effect. Silencing of CD39 promoted the expression of ATP and P2Y2R. These results indicate that CD39 attenuates alcohol-induced steatohepatitis by scavenging extracellular ATP to indirectly regulate the expression of P2Y2R. Interestingly, P2Y2R paradoxically boosts CD39 activity. Thus, blockade of the extracellular ATP-P2Y2R signalling represents a potential therapeutic approach against alcoholic liver disease, and CD39 is a potential therapeutic target.

Histopathological Features and Composition of Gut Microbiota in Rhesus Monkey of Alcoholic Liver Disease.

Alcohol-induced chronic liver disease (ALD) is becoming the most common liver disease in the world. However, there are no effective, universally accepted therapies for ALD. The etiology of ALD remains blurry so far. Historical evidence has demonstrated a link between the liver and gut microbiota. But it is difficult to distinguish the effect of gut microbiota changes caused by alcohol consumption in humans since the microbiota change detected in humans is complicated by diet and environmental factors. Due to the genetic, physiological, metabolic, and behavioral similarities to humans, the rhesus monkey provides excellent translational validity in preclinical studies, and the diet and environmental conditions can be controlled well in rhesus monkey. In our study, we explored the relationship between ALD and the gut microbiome in the rhesus monkeys with alcoholic liver steatosis. Our results showed that there was a change of the bacterial community structure in monkeys with ALD. Differences of the relative abundances of gut microbiota at phylum, order, family, genus, and species levels were observed between control monkeys and monkeys with ALD, and different pathways enriched in the monkeys with ALD were identified by metagenomic function analysis. Firmicutes, Proteobacteria, Verrucomicrobia tended to increase whereas Bacteroidetes and Actinobacteria decreased in the fecal microbiota of ALD group compared to the control group. Lactobacillales and Lactobacillus significantly decreased in ALD monkeys compared with normal monkeys, Streptococcus was lower in the ALD group compared with the control group. The non-human primate model of ALD will be useful for exploration of the microbiome markers as diagnosis and potentially prognosis for ALD. The ALD model will benefit the development of new therapeutic procedures for treating ALD and provide safety and efficacy evaluation for clinical application.

Mulberry and dandelion water extracts prevent alcohol-induced steatosis with alleviating gut microbiome dysbiosis.

Chronic alcohol intake causes hepatic steatosis and changes the body composition and glucose metabolism. We examined whether water extracts of mulberry (WMB) and white flower dandelion ( Taraxacum coreanum Nakai, WTC) can prevent and/or delay the symptoms of chronic ethanol-induced hepatic steatosis in male Sprague Dawley rats, and explored the mechanisms. Ethanol degradation was examined by orally administering 3 g ethanol/kg bw after giving them 0.3 g/kg bw WMB or WTC. All rats were continuously provided about 7 g ethanol/kg bw/day for four weeks and were given either of 0.1% dextrin (control), WMB, WTC, or water extracts of Hovenia dulcis Thunb fruit (positive-control) in high-fat diets. Area under the curve of serum ethanol levels was lowered in descending order of control, WTC and positive-control, and WMB in acute ethanol challenge. WMB and WTC prevented alcohol intake-related decrease in bone mineral density and lean body mass compared to the control. After glucose challenge, serum glucose levels increased more in the control group than other groups in the first part and the rate of decrease after 40 min was similar among all groups. These changes were associated with decreasing serum insulin levels. WMB had the greatest efficacy for decreasing triglyceride and increasing glycogen deposits. WMB and WTC prevented the disruption of the hepatic cells and nuclei while reducing malondialdehyde contents in rats fed alcohol, but the prevention was not as much as the normal-control. The ratio of Firmicutes to Bacteroidetes in the gut was much higher in the control than the normal-control, but WTC and WMB decreased the ratio compared to the control. WMB and WTC separated the gut microbiota community from the control. In conclusion, WMB and WTC protected against alcoholic liver steatosis by accelerating ethanol degradation and also improved body composition and glucose metabolism while alleviating the dysbiosis of gut microbiome by chronic alcohol intake. Impact statement Excessive alcohol consumption is associated with serious pathologies and is common in much of the world. Pathologies include liver damage, glucose intolerance, and loss of lean body mass and bone mass. These pathologies are mediated by changes in metabolism as well as toxic metabolic byproducts, and possibly by gut dysbiosis. In this study, we demonstrate that aqueous extracts of mulberry and dandelion protected rats against ethanol-induced losses in lean body and bone masses, improved glucose tolerance and partially normalized gut bacterial populations, with mulberry extract being generally more effective. This research suggests that mulberry and dandelion extracts may have the potential to improve some of the pathologies associated with excess alcohol consumption, and that further clinical research is warranted.

Amelioration of Alcoholic Liver Steatosis by Dihydroquercetin through the Modulation of AMPK-Dependent Lipogenesis Mediated by P2X7R-NLRP3-Inflammasome Activation.

Dihydroquercetin (TAX) is the most abundant dihydroflavone found in onions, milk thistle, and Douglas fir bark. We investigated whether TAX could inhibit lipid accumulation in alcoholic liver steatosis in vivo and in vitro. An in vivo model was established by intragastrically treating mice with ethanol, and an in vitro model was created by treating HepG2 cells with ethanol. TAX regulated SREBP1 and ACC expression by elevating LKB1 and AMPK phosphorylation. Also, TAX upregulated SIRT1 expression, which was suppressed by ethanol intake. Decreased expression of P2X7R and NLRP3 and suppressed cleavage of caspase-1 by TAX resulted in the inhibition of IL-1β production and release. Additionally, TAX reduced lipogenesis and promoted lipid oxidation via the regulation of AMPK and ACC in ethanol-treated steatotic HepG2 cells. TAX downregulated IL-1β cleavage responses to LPS and ATP stimulation in HepG2 cells. P2X7R deficiency attenuated lipid accumulation, characterized by increased AMPK activity and decreased SREBP1 expression in ethanol-treated HepG2 cells. Our data showed that TAX exhibited the ability to inhibit lipogenesis and a hepatoprotective capacity, indicating that TAX has therapeutic potential for preventing alcoholic liver steatosis.

From the Editor’s desk....: May 2018

From the Editor’s desk....: May 2018

Controlled attenuation parameter and alcoholic hepatic steatosis: Diagnostic accuracy and role of alcohol detoxification

Controlled attenuation parameter (CAP) is a novel non-invasive measure of hepatic steatosis, but it has not been evaluated in alcoholic liver disease. Therefore, we aimed to validate CAP for the assessment of biopsy-verified alcoholic steatosis and to study the effect of alcohol detoxification on CAP. This was a cross-sectional biopsy-controlled diagnostic study in four European liver centres. Consecutive alcohol-overusing patients underwent concomitant CAP, regular ultrasound, and liver biopsy. In addition, we measured CAP before and after admission for detoxification in a separate single-centre cohort. A total of 562 patients were included in the study: 269 patients in the diagnostic cohort with steatosis scores S0, S1, S2, and S3 = 77 (28%), 94 (35%), 64 (24%), and 34 (13%), respectively. CAP diagnosed any steatosis and moderate steatosis with fair accuracy (area under the receiver operating characteristic curve [AUC] ≥S1 = 0.77; 0.71-0.83 and AUC ≥S2 = 0.78; 0.72-0.83), and severe steatosis with good accuracy (AUC S3 = 0.82; 0.75-0.88). CAP was superior to bright liver echo pattern by regular ultrasound. CAP above 290 dB/m ruled in any steatosis with 88% specificity and 92% positive predictive value, while CAP below 220 dB/m ruled out steatosis with 90% sensitivity, but 62% negative predictive value. In the 293 patients who were admitted 6.3 days (interquartile range 4-6) for detoxification, CAP decreased by 32 ± 47 dB/m (p <0.001). Body mass index predicted higher CAP in both cohorts, irrespective of drinking pattern. Obese patients with body mass index ≥30 kg/m2 had a significantly higher CAP, which did not decrease significantly during detoxification. CAP has a good diagnostic accuracy for diagnosing severe alcoholic liver steatosis and can be used to rule in any steatosis. In non-obese but not in obese, patients, CAP rapidly declines after alcohol withdrawal. CAP is a new ultrasound-based technique for measuring fat content in the liver, but has never been tested for fatty liver caused by alcohol. Herein, we examined 562 patients in a multicentre setting. We show that CAP highly correlates with liver fat, and patients with a CAP value above 290 dB/m were highly likely to have more than 5% fat in their livers, determined by liver biopsy. CAP was also better than regular ultrasound for determining the severity of alcoholic fatty-liver disease. Finally, we show that three in four (non-obese) patients rapidly decrease in CAP after short-term alcohol withdrawal. In contrast, obese alcohol-overusing patients were more likely to have higher CAP values than lean patients, irrespective of drinking.

Molecular-genetic risk factors of non-alcoholic fatty liver disease and obesity in patients with arterial hypertension

The objective: to analyze the role of polymorphic variants of the Peroxisome Proliferator-Activated Receptor-γ2 (PPAR-γ2, Pro12Ala) and angiotensin converting enzyme (ACE, I/D) genes as possible causative risk factors of Non-alcoholic Fatty Liver Disease (NAFLD), Abdominal Obesity (AO), Diabetes Mellitus type 2 (DM 2) in essential arterial hypertension (EAH) patients.Materials and methods. 96 patients with non-alcoholic fatty liver disease (NAFLD), EAH II stage, of 1–2 degrees of blood pressure, high and very high risk with concomitant AO participated in the research: 41,67% (40) men and 58,33% (56) women, the average age was 53,70±5,34 years. The function of the liver was studied by the activity of organ specific enzymes. The polymorphism of PPAR-γ2 (Pro12Ala) and АСЕ (I/D) genes were studied by the PCR based method. The control group included 50 practically healthy subjects.Results. Among the residents of the Northern Bukovyna suffering from NAFLD, AO and EAH the deletion in the 16th intron of the АСЕ (rs 4646994) gene in the homozygous condition occurs in 32,29% of cases, which is by 14.29% more frequently than in the control group sunbjects (χ2=3,38; р=0,048). The DD genotype presence of the АСЕ gene almost double increases the relative risk of NAFLD in general [OR=2,17; р=0,048] and non-alcoholic liver steatosis in particular [OR=2,32; р=0,038], as well as CD2 type [OR=3,46; p=0,015].The PPAR-γ2 (rs1801282) gene genotypes distribution between patients with NAFLD and control group subjects didn’t significantly differ with the prevalence of Pro alleles 3,85 and 6,14 times (p<0.001). The minor Ala-allele presence of the PPAR-γ2 gene (rs1801282) in the genotype increases the relative risk of non-alcoholic steatohepatitis 1.73 times [OR=2,67; p=0,054], however, plays a protective role in the severe obesity degrees appearance [OR=0,29; p=0,008]. The homozygous Pro allele carrier, on the contrary, increases the likelihood of AO ІІ and ІІІ degrees more than 3 times [OR=3,41; p=0,016].Conclusion. The presence of DD genotype of the ACE (rs4646994) gene in patients with EAH is a risk factor of NAFLD in general and non alcoholic liver steatosis in particular, as well as DM 2. The presence of the minor Ala allele of PPAR-γ2 gene (rs1801282) increases the relative risk of non alcoholic steatohep atitis, and the ProPro genotype increases the chances for AO II and III degree appearance.

Intestinal Alkaline Phosphatase Attenuates Alcohol-Induced Hepatosteatosis in Mice.

Bacterially derived factors from the gut play a major role in the activation of inflammatory pathways in the liver and in the pathogenesis of alcoholic liver disease. The intestinal brush-border enzyme intestinal alkaline phosphatase (IAP) detoxifies a variety of bacterial pro-inflammatory factors and also functions to preserve gut barrier function. The aim of this study was to investigate whether oral IAP supplementation could protect against alcohol-induced liver disease. Mice underwent acute binge or chronic ethanol exposure to induce alcoholic liver injury and steatosis±IAP supplementation. Liver tissue was assessed for biochemical, inflammatory, and histopathological changes. An ex vivo co-culture system was used to examine the effects of alcohol and IAP treatment in regard to the activation of hepatic stellate cells and their role in the development of alcoholic liver disease. Pretreatment with IAP resulted in significantly lower serum alanine aminotransferase compared to the ethanol alone group in the acute binge model. IAP treatment attenuated the development of alcohol-induced fatty liver, lowered hepatic pro-inflammatory cytokine and serum LPS levels, and prevented alcohol-induced gut barrier dysfunction. Finally, IAP ameliorated the activation of hepatic stellate cells and prevented their lipogenic effect on hepatocytes. IAP treatment protected mice from alcohol-induced hepatotoxicity and steatosis. Oral IAP supplementation could represent a novel therapy to prevent alcoholic-related liver disease in humans.

Blog educativo bioquímicativa: uma forma de vivenciar a educação em bioquímica e sua relação com a fisiopatologia

O projeto de extensão BioquímicAtiva surgiu com o intuito de utilizar a internet como um meio de ensino da bioquímica, dada a dificuldade que os alunos comumente relatam sobre a compreensão dessa disciplina e sua associação com a prática profissional. Por esta razão, o presente trabalho avaliou a eficiência de um blog educativo sobre bioquímica, elaborado a partir de um post de fácil linguagem, didático e interativo, utilizando-se da interdisciplinaridade com a fisiopatologia. Toda a criação do blog e de seu conteúdo foi executada pelos integrantes do projeto, os quais se basearam na literatura científica para criação do post ‘’Você sabe o que é Esteatose hepática alcoólica?’’. Para avaliação do blog considerou-se parâmetros de visão geral e informações demográficas, gerados pelo Google Analytics. O conteúdo postado foi avaliado pela enquete “O que você achou deste conteúdo?”, disponível ao final do post para votação. O blog foi avaliado por quinze dias, contados a partir da data de sua divulgação, com registro total de 1236 visualizações nesse período, incluindo usuários de diversas cidades brasileiras e alguns países. Foram notificados 26 votos, sendo 25 para a opção “Excelente” (96,15%), um para “Bom” (3,84%) e nenhum voto para “indiferente”. Conclui-se que o projeto elaborado foi positivo, o que demonstra a importância do uso de novos métodos de ensino e tecnologias que possam contribuir para a formação e interesse pela bioquímica, bem como estimular seu uso para outros cursos de graduação, melhorando assim o processo de ensino e aprendizagem dos acadêmicos.