Stages Of Liver Damage Research Articles

Spatial Omics Reveals that Cancer-Associated Glycan Changes Occur Early in Liver Disease Development in a Western Diet Mouse Model of MASLD.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a progressive disease and comprises different stages of liver damage; it is significantly associated with obese and overweight patients. Untreated MASLD can progress to life-threatening end-stage conditions, such as cirrhosis and liver cancer. N-Linked glycosylation is one of the most common post-translational modifications in the cell surface and secreted proteins. N-Linked glycan alterations have been established to be signatures of liver diseases. However, the N-linked glycan changes during the progression of MASLD to liver cancer are still unknown. Here, we induced different stages of MASLD in mice and liver-cancer-related phenotypes and elucidated the N-glycome profile during the progression of MASLD by quantitative and qualitative profiling in situ using matrix-assisted laser desorption ionization (MALDI) imaging mass spectrometry (IMS). Importantly, we identified specific N-glycan structures including fucosylated and highly branched N-linked glycans at very early stages of liver injury (steatosis), which in humans are associated with cancer development, establishing the importance of these modifications with disease progression. Finally, we report that N-linked glycan alterations can be observed in our models by MALDI-IMS before liver injury is identified by histological analysis. Overall, we propose these findings as promising biomarkers for the early diagnosis of liver injury in MASLD.

Recent insight into the role of macrophage in alcohol-associated liver disease: a mini-review

Alcohol-associated liver disease (ALD) is a condition that develops due to prolonged and excessive alcohol consumption. It encompasses various stages of liver damage, including fatty liver, alcoholic hepatitis, and cirrhosis. Immune cells, particularly macrophages, of various types play a significant role in the onset and progression of the disease. Macrophages observed in the liver exhibit diverse differentiation forms, and perform a range of functions. Beyond M1 and M2 macrophages, human macrophages can polarize into distinct phenotypes in response to various stimuli. Recent advancements have improved our understanding of macrophage diversity and their role in the progression of ALD. This mini-review provides a concise overview of the latest findings on the role and differentiation of macrophages in ALD. Additionally, it discusses potential therapeutic targets associated with macrophages and explores potential therapeutic strategies.

Сhanges in the composition of gut microbiota in patients with chronic hepatitis С, non-alcoholic fatty liver disease at different stages of liver disease: a review

INTRODUCTION. The human gut microbiota is a set of microorganisms that are in symbiosis with the host organism. The predominance of bacteria of Bacteroidetes, Firmicutes, Actinobacteria types is a characteristic feature of human gut microbiota. However, the composition of the gut microbiota is subject to change under the influence of a number of factors. The influence of the gut-liver axis on the pathogenesis of many chronic liver diseases of various etiologies, both infectious and metabolic, has been proven. At the same time, in the analyzed literature, the data on the relationship between the nature of GM changes and the stage of liver damage are quite controversial.
 OBJECTIVE. Updating and systematization of ideas about the nature of changes in the composition of intestinal microbiota in chronic hepatitis C (CHC) and non-alcoholic fatty liver disease (NAFLD) at different stages of the disease.
 MATERIALS AND METHODS. We analyzed the scientific literature on GM changes in chronic liver diseases with regard to their etiology and disease stage, in particular in chronic viral hepatitis C and NAFLD. The search was conducted in PubMed, Google Scholar, and eLIBRARY databases for 2000-2023 using the following keywords: gut microbiota; microbiota in hepatitis C; microbiota in NAFLD; microbiota in liver fibrosis; liver fibrosis progression. Sixty scientific articles were analyzed and 42 sources were selected, of which more than 60% were published within the last five years.
 RESULTS. This review presents the results of original studies describing the main taxonomic differences between the microbiomes of healthy individuals and patients with CHC complicated by the formation of liver cirrhosis and hepatocellular carcinoma (HCC). Thus, patients with viral cirrhosis showed impoverishment of intestinal commensals Lachnospiraceae and Ruminococcaceae and significant enrichment of Streptococcaceae, Lactobacillaceae and Enterobacteriaceae. In patients with non-alcoholic steatohepatitis (NASH) and FII, enrichment of GM Phylum Bacteroides and Proteobacteria, Families Lachnospiraceae and Enterobacteriaceae, Genera Blautia and Escherichia, with a concomitant decrease of Prevotella was detected.
 DISCUSSION. The analysis of GM taxonomic units in the previously described studies did not reveal significant differences depending on the etiology of the underlying disease. However, a decrease in the representative of the phylum Firmicutes - Ruminococcaceae was observed in CHC in contrast to NAFLD. A more distinct relationship can be observed in the changes of intestinal microbiota in F(III-IV) irrespective of etiology. Thus, in advanced liver fibrosis and cirrhosis of both CHC and NAFLD etiologies, enrichment of GM has been described due to an increase in phylum Proteobacteria, in particular the genus Escherichia and phylum Firmicutes (Blautia, Veillonellaceae). In the studies analyzed, at F(III-IV), an enrichment of the microbiota is observed due to an increase in the phylum Bacteroidetes. Regarding the genus Prevotella, contradictory results were found. In addition, there are ambiguous data regarding the genera Lactobacillus, Ruminococcaceae and Bifidobacteria.
 CONCLUSION. Based on the results of the literature review, we obtained contradictory data on GM composition in patients with CHC and NAFLD. This may be due to the size of samples, heterogeneity of initial data during the selection of patients in the study. The relationship between the composition of microbiota and persistent inflammatory process, expressed liver fibrosis is observed. GM reflects not only functional disorders in chronic liver diseases of various etiologies, but can also serve as a diagnostic indicator of their progression.

Evaluation of Small Intestinal Permeability in Patients with Overlap Syndrome (Autoimmune Hepatitis/Primary Biliary Cholangitis)

Аim: to evaluate the state of small intestine permeability by the “double sugar test” in patients with overlap syndrome (autoimmune hepatitis / primary biliary cholangitis (AIH / PBC)).Materials and methods. Prospectively, 56 people were included in the study. Of these, 26 were diagnosed with AIH/PBC, 30 were in the control group. The diagnosis was made in accordance with the current recommendations. The average age of patients was 49.7 ± 13.8 years, healthy volunteers — 48.6 ± 9.2 years. The determination of the permeability of the small intestine was carried out by a “double sugar test” (the ratio of lactulose/mannitol in urine), using the method of high-performance liquid chromatography — mass spectrometry.Results. In patients with AIH/PBC, an increase in intestinal permeability was found — 0.20 [0.09; 0.30] (p < 0.001) compared with the control group 0.01 [0.01; 0.02]. We divided patients at the stage of liver damage. An increased small intestinal permeability was revealed: hepatitis stage — 0.19 [0.13; 0.30] (p < 0.001), liver cirrhosis stage — 0.18 [0.09; 0.30] (p < 0.05) compared with the control group. In the early stages of disease (1 month from the onset of the disease) had an increased lactulose/mannitol ratio — 0.13 [0.05; 0.26] (p < 0.001) compared to the control group. In the presence of portal hypertension (PH), small intestinal permeability was increased — 0.18 [0.09; 0.30] (p < 0.001) compared with the control group.Conclusions. An increase in small bowel permeability was found in patients with overlapping syndrome. All patients had increased intestinal permeability (regardless of the presence of extrahepatic manifestations).

IL-1\u03b2-activated mTORC2 promotes accumulation of IFN-\u03b3+ \u03b3\u03b4 T cells by upregulating CXCR3 to restrict hepatic fibrosis

Liver fibrosis represents a severe stage of liver damage, with hallmarks of inflammation, hepatic stellate cell activation, and extracellular matrix accumulation. Although previous studies demonstrated γδ T cells are involved in liver fibrosis, the precise role and mechanisms of γδ T cells migrating to fibrotic liver have not been elucidated. Here, we aim to investigate the functional subsets of γδ T cells in hepatic fibrosis and to further explore the underlying causes and drivers of migration. In this study, we observed that γδ T cells accumulate in fibrotic liver. Adoptive transfer of γδ T, especially Vγ4 γδ T subset, can significantly alleviate liver fibrosis. In addition, CCl4 treatment also leads to activation of mTOR signaling in γδ T cells. Genetic deletion of the Rictor gene, but not Raptor, in γδ T cells markedly exacerbated liver fibrosis. Mechanistically, CCl4-induced liver injury causes macrophage accumulation in the liver, and IL-1β produced by macrophages promotes mTORC2 signaling activation in γδ T cells, which upregulates T-bet expression and eventually promotes CXCR3 transcription to drive γδ T cell migration. Moreover, hepatic γδ T cells ameliorated liver fibrosis by cytotoxicity against activated hepatic stellate cells in FasL-dependent manner, and secrete IFN-γ to inhibit the differentiation of pro-fibrotic Th17 cells. Thus, IL-1β-activated mTORC2 signaling in γδ T cells upregulates CXCR3 expression, which is critical for IFN-γ+ γδ T cells migration into the liver and amelioration of liver fibrosis. Our findings indicate that targeting the mTORC2 or CXCR3 in γδ T cells could be considered as a promising approach for γδ T cell immunotherapy against liver fibrosis.

1164-P: A Zebrafish Nonalcoholic Fatty Liver Disease Model Links Liver Inflammation with Pancreatic Inflammation and Hyperglycemia

The spectrum of nonalcoholic liver disease (NAFLD) ranges from simple steatosis to steatohepatitis with the latter having the potential to irreversibly progress to cirrhosis. Compared with the general population, NAFLD is more frequently seen in patients with type 2 diabetes (T2D), and conversely, NAFLD is associated with increased incidence of T2D. Existing rodent models of NAFLD focus narrowly on certain stages of liver damage and insufficiently encompass all metabolic changes, limiting their utility. The aims of this study were to develop a new model of NAFLD in young zebrafish and use it to probe the associations between liver inflammation and pancreatic islet function. We examined hepatopancreatic inflammation under three conditions: (1) in a zebrafish line (fabp10-CETI-PIC3) in which liver-specific expression of pro-inflammatory cytokines IL-1β, TNFα, and IFN-γ was induced by doxycycline treatment (M1); (2) in wild-type zebrafish fed a high-fat diet (M2); and (3) in a combination of these models (M3). All three conditions showed increased expression of inflammatory cytokines compared to Tg(fabp10:cre) control larvae, and when compared to controls, all three showed increased macrophage infiltration in the liver. Strikingly, staining with CellRox Deep Red to assess reactive oxygen species (ROS)-mediated stress in the liver revealed a 2-fold increase in the M3 combination model, but not in either M1 or M2. Further, although all models showed hyperglycemia, decreased insulin transcription without β cell death, and ROS accumulation in the pancreas, only the M3 condition demonstrated a significant increase in macrophage infiltration of the pancreatic islets as compared to controls. In sum, our novel zebrafish model of liver steatosis and steatohepatitis is a promising model in which to study the intricate associations between NAFLD and T2D, and may serve as an amenable platform for screening novel pharmaceutical interventions. Disclosure S. Ibrahim: None. I. Haider: None. A. Kulkarni: None. I. Doycheva: None. R. G. Mirmira: Advisory Panel; Self; Hibercell Inc., Sigilon Therapeutics, Inc., Veralox Therapeutics, Employee; Spouse/Partner; Beta Bionics, Inc. E. K. Sims: None. R. Anderson: None.

SPECIAL METHODS OF DIAGNOSIS OF THE FUNCTIONAL CONDITION OF THE LIVER IN PATIENTS WITH CIRRHOSIS

Abstract. The aim. Using special methods to investigate the functional state of the liver in patients with cirrhosis.
 Materials and methods. The study involved 95 patients with LC who were hospitalized from 2018 to 2020 in the Department of Anesthesiology and Intensive Care, Surgical and Gastroenterological Departments of the Transcarpathian Regional Clinical Hospital. Andriy Novak (Uzhhorod). All patients underwent C13-metacetin breath test (C13-MDT) to determine the degree of liver damage. To determine the degree of fibrosis in the examined patients used the following specially developed tests: Forns test, FibroIndex test, FIB-4 test (Fibrosis 4 Score), APRI test (AST-to-Platelet Ratio Index), MDA test (multivariate discriminant analysis), test GUCI (Gothenburg University Cirrhosis Index), FPI test.
 Results. In all examined patients with LC on the results of C13-MDT revealed changes characteristic of liver cirrhosis. In group I of patients with LC, the percentage of functioning hepatocytes was 54.7 ± 5.3%. MFG was the lowest in group III patients with LC - 18.2 ± 1.8% and was statistically significantly different from patients in group I (p <0.05). A statistically significant difference between MFG parameters was also found between groups I and II of patients with LC - p <0.05. We found a strong correlation between the results of C13-MDT and histostructure disorders and the stage of liver damage (mainly F4) by METAVIR in the examined patients (r = 0.72; p <0.01). Based on the results of statistical analysis, it was found that the most effective test based on serum biochemical parameters is the FPI test.
 Conclusions. The most pronounced changes that characterize the reduced functional capacity of the liver, found in patients of group III - p <0,05. The most effective test based on serum biochemical parameters is the FPI test. Severe liver cirrhosis is characterized by higher plasma levels of endothelial mediators, fibrinolysis regulators and lower platelet counts, levels of physiological anticoagulants, prolonged invertebrate values.

ABCB1, CYP2B6, and CYP3A4 genetic polymorphisms do not affect methadone maintenance treatment in HCV-positive patients.

The aim of this study was to determine the influence of ABCB1, CYP2B6, and CYP3A4 genetic polymorphisms on methadone metabolism in patients with hepatitis C virus (HCV) undergoing methadone maintenance treatment (MMT). The study included 35 participants undergoing MMT, who were divided in three groups: HCV-positive (N=12), HCV-negative (N=16), and HCV clinical remission (CR) (N=7). The concentrations of methadone and its main metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) were determined with gas chromatography-mass spectrometry. The patients were genotyped for ABCB1 rs1045642, CYP2B6 rs3745274, CYP3A4 rs2242480, and CYP3A4 rs2740574 polymorphisms. Differences between single nucleotide polymorphism (SNP) genotypes and methadone-to-EDDP ratio were analysed with one-way ANOVA, which showed no significant difference between the genes (p=0.3772 for ABCB1 rs1045642, p=0.6909 for CYP2B6 rs3745274, and p=0.6533 for CYP3A4 rs2242480). None of the four analysed SNP genotypes correlated with methadone-to-EDDP concentration ratio. A major influence on it in hepatitis C-positive patients turned out to be the stage of liver damage.

Non-alcoholic fatty liver disease and left ventricular diastolic dysfunction in patients with metabolic syndrome

The purpose of the study was to determine the frequency and nature of manifestations of left ventricular diastolic dysfunction in patients with non-alcoholic fatty liver disease with the background of metabolic syndrome (MS).Characteristics of patients and research methods: To solve this problem, 227 patients with MS (130 men and 97 women) were examined. The study was conducted in two phases. At the first stage, all patients with MS are divided into two groups: patients with NAFLD — 205 people and 22 — without liver damage. At the next stage, a group of NAFLD patients with left ventricular DD (n=136) (66.34%) (the main group) and patients with no DD (n=69) (33.66%) (the comparison group) were identified.Results of the study: the results of diagnostics, clinical and laboratory manifestations of left ventricular diastolic dysfunction in 136 patients with NAFLD on the background of metabolic syndrome are Presented. Significant factors of development and progression of diastolic dysfunction were identified, which include increased body weight and adipose tissue, insulin resistance, dyslipidemia, hyperuricemia, activity and stage of liver damage. According to the data of structural and functional changes in the myocardium, the role of diastolic dysfunction in the development of chronic heart failure in patients with NAFLD with manifestations of metabolic syndrome has been established.Conclusion: it was Found that the development of left ventricular diastolic dysfunction in patients with NAFLD is due to the morphofunctional state of the liver.

Nutritional status at Wilson’s disease and its effect on oxidative stress

Aim . To study characteristic features of nutritional status of patients with Wilson ’s disease depending on stage of liver damage and determine the impact of malnutrition on oxidative stress level. Methods . The study included 73 subjects, 33 (15 men and 18 women) had a confi rmed diagnosis of Wilson’s disease. All patients with Wilson’s disease (the study group) had liver damage: non-cirrhotic stages — 12 (36.3%), cirrhosis — 21 (63.6%). The control group consisted of 40 healthy subjects (20 men and 20 women) with neither body weight defi ciency, nor obesity, nor liver disease. All patients were conducted standard blood tests (clinical and biochemical blood tests), measuring malonodialdehyde (MDA) in blood serum by high-performance liquid chromatography, as well as assessment of height, weight and BMI (Body Mass Index) and analysis the body mass composition with bioelectricalimpedanceanalyzer “Diamant AIST”. Results . 1. Malnutrition was more common among patients with liver cirrhosis (28.6%) than in patients with non-cirrhotic stages (8.3%). 2. The nutritional status of patients with liver cirrhosis was signifi cantly diff erent from the control group: BMI, total protein and albumin levels, lymphocytes and arm muscular circumference were signifi cantly lower in patients with liver cirrhosis compared to the control group. The decrease in fat and muscular mass was more pronounced among women than men with liver cirrhosis. 3. The group of patients with chronic hepatitis did not significantly statistically diff er from the control group in most nutritional status parameters, except for albumin level, which was signifi cantly lower in persons of both sexes with noncirrhotic stages; total protein level and body fat percent were signifi cantly higher in men with non-cirrhotic stages than in the control group. 4. The marker of oxidative stress level was signifi cantly higher in patients with liver cirrhosis, and was higher in patients with malnutrition.

Structural and Morphological Changes in the Liver Due to Intestinal Endotoxins.

Under normal physiological conditions, endotoxin (ET) released during self-renewal of the colibacillus pool is an obligate stimulus for the formation of the immune system and homeostasis of the body. Violation of the barrier function of the intestinal wall and the mechanisms of neutralization of endotoxin lead to systemic endotoxemia of intestinal origin. Its development is facilitated by stress, intoxication, a decrease in nonspecific resistance of the body, as well as damage to the intestinal mucosa and dysbiosis, where the mucous membrane is more vulnerable and permeable to endotoxin. The aim of this study is to compare and assess the severity and nature of hepatocyte damage from endotoxin exposure and the degree of manifestation of stress due to oxidation, to determine the characteristics of structural changes in hepatocytes and to assess the oxidation stress during endotoxin intoxication in the experiment with biochemical markers. The experiments were conducted on 40 non-linear rats, divided into two groups of 20 animals. Group 1 animals received intraperitoneal injections of ET of Escherichia coli drug (Sigma USA K-235) for seven days at a rate of 0.1 mg/kg of the body weight. Animals of the second group served as the control group. Character and stage of liver damage were studied using morphological methods, including electron and light microscopy. In studying oxidizing stress, biochemical methods were used to define the changes, such as conjugated dienes and dienketones, spontaneous oxidizing modification of proteins. 1. The severity and depth of morphological changes in the liver during endotoxin intoxication were correlated with the dynamics of the content of lipid oxidation products (CD and DK, MDA) and proteins. There was a tendency for a more significant increase in the oxidative modification of proteins in serum. This confirms the data on the primary damage of proteins by free radicals. 2. When exposed to intestinal microflora endotoxin, pronounced dyscirculatory changes, fatty and hydropic degeneration of hepatocytes with signs of toxic damage to their nuclei were determined, but at the same time, the increased hyperplastic activity of sinusoidal cells remained associated with the effects of endotoxin. These changes are associated with both the direct toxic effect of endotoxin, and the effects of oxidative stress, in which endotoxin is a potent inducer.

CHANGES IN THE LEVEL OF α1-ANTITRYPSIN IN PATIENTS WITH NON-ALCOHOLIC FATTY LIVER DISEASE

The aim: To investigate changes in the level of α1-antitrypsin (A1AT) in blood and in stool and their diagnostic value in patients with NAFLD and with impaired carbohydrate metabolism at different stages of liver damage. Materials and methods: 34 patients with non-alcoholic fatty hepatosis (NAFH) and 40 patients with non-alcoholic steatohepatitis (NASH) were examined. Enzyme-linked immunosorbent assay for all patients with serum and coli was performed by determining the level of A1AT. Results: In patients with NAFLD, in combination with IR, the level of A1AT in the blood plasma is only 1.6 times higher than that of the control group (p <0.05), and in the case of combination of NAFH and type 2 diabetes mellitus (T2DM) - it is 2 times higher (p <0.05). In patients with NASH in combination with insulin resistance (IR), the level of A1AT in the serum was 4.1 times higher than in the control group, and for NASH and type 2 diabetes - 4.2 times. Conclusions:The level of A1AT in the blood plasma increases proportionally to the degree of progression of liver damage (from the minimum values at NAFH to the maximum at NASH), regardless of the type of carbohydrate metabolism disturbance. The combination of NAFLD and T2DM is accompanied by a more pronounced increase in A1AT in stool and α1-antitrypsin clearance than the combination of NAFLD and IR.

Alteration of splicing factors' expression during liver disease progression: impact on hepatocellular carcinoma outcome.

Trans-acting splicing factors (SF) shape the eukaryotic transcriptome by regulating alternative splicing (AS). This process is recurrently modulated in liver cancer suggesting its direct contribution to the course of liver disease. The aim of our study was to investigate the relationship between the regulation of SFs expression and liver damage. The expression profile of 10 liver-specific SF and the AS events of 7 genes associated with liver disorders was assessed by western-blotting in 6 murine models representing different stages of liver damage, from inflammation to hepatocellular carcinoma (HCC). Relevant SFs (PSF, SRSF3, and SRSF6) and target genes (INSR, SRSF3, and SLK) modulated in mice were investigated in a cohort of 179 HCC patients. Each murine model of liver disease was characterized by a unique SF expression profile. Changes in the SF profile did not affect AS events of the selected genes despite the presence of corresponding splicing sites. In human HCC expression of SFs, including the tumor-suppressor SRSF3, and AS regulation of genes studied were frequently upregulated in tumor versus non-tumor tissues. Risk of tumor recurrence positively correlated with AS isoform of the INSR gene. In contrast, increased levels of SFs expression correlated with an extended overall survival of patients. Dysregulation of SF expression is an early event occurring during liver injury and not just at the stage of HCC. Besides impacting on AS regulation, overexpression of SF may contribute to preserving hepatocyte homeostasis during liver pathogenesis.

Protective effect of anthocyanin on alcohol‐induced liver fibrosis is due to the regulation on AMPK/mTOR/autophagy pathway and energy status

Liver fibrosis is an advanced stage of liver damage by alcohol. We hypothesize that anthocyanin can protect alcohol induced liver fibrosis. We fed mice with high fat diet with ehanol in dringking water to induce liver fibrosis. Anthocyanin was supplied by gavage every day. We found that 12 wks feeding of high fat diet and alcohol induced lipid accumulation and increased expression of collagen α, α‐SMA, TGF‐β, and TIMP‐1 and −2, while anthocyanin supplement inhibited the increase of these parameters. In addition, we found that high fat and ethanol feeding induced increase of liver oxidative stress, increased activation of AMPK pathway, and increased autophagy, while anthocyanin supplement inhibited these effects in liver. Moreover, we found that supplement of anthocyanin reversed energy deficiency induced by high fat and ethanol feeding, including the levels of ATP, ADP, AMP, NAD+, and NMN in the liver. In conclusion, the protective effect of anthocyanin on ethanol induced liver fibrosis is due to the regulation on AMPK/mTOR/autophagy pathway, which finally regulates the energy supply in the liver.Support or Funding InformationNational Natural Science Funding (81370528, 81573142)(Yang, L), Provincial Natural Science Funding of Guangdong (2015A030313078) (Yang, L.)

Influence of cytokine and cytokine receptor gene polymorphisms on the degree of liver damage in patients with chronic hepatitis C

Hepatic fibrosis may be the result of repetitive injury to hepatocytes caused by HCV infection and the immune response to it. Cytokines regulate the inflammatory response to injury and modulate hepatic fibrogenesis. Single nucleotide polymorphisms (SNPs) located in cytokine genes may influence the cytokine expression and secretion that may contribute to hepatic fibrogenesis in HCV infection. The aim of this study was to determine the genotype of 22 SNPs found in the genes of 13 cytokines/cytokine receptors to assess the influence of polymorphic variants on the stage of liver damage in Brazilian patients chronically infected with HCV genotype 1 only. 141 unrelated patients were grouped according to their stage of fibrosis: absence of fibrosis or patients in the initial stages of fibrosis (F0-F2, n=84), patients with advanced stages of fibrosis or cirrhosis (F3-F4, n=57), without cirrhosis (F0-F3, n=103), and with cirrhosis (F4, n=38). The comparison of frequencies in each sub-sample was performed by 2×2 contingency tables using the chi-square or Fisher's exact test. Stepwise logistic regression was also used to assess independent associations between cirrhosis or fibrosis with polymorphic variants. The TNFA-308G:A genotype conferred increased risk of fibrosis and cirrhosis. The TNFA-238G:G genotype was associated with protection from cirrhosis. The IL10-819C:T genotype conferred protection from fibrosis and the IL1B-511C:T genotype conferred increased risk of cirrhosis. Some of these genotypes showed results on the borderline of statistical significance in the bivariate analysis. We conclude that gene variants of cytokines/receptors may influence liver damage in patients chronically infected by HCV genotype 1.

Protective effect of Thymic Humoral Factor on porcine serum-induced hepatic fibrosis and liver damage in Wistar rats

Introduction. Immunomodulatory drugs have been reported to have anti-inflammatory and anti-fibrotic properties. Thymic Humoral Factor (THF), a peptide produced in the thymus, causes a potent immunomodulatory effect on different components of the immune system. Objective. To evaluate the effect of THF on different stages of liver damage and fibrosis induced in rats through the administration of porcine serum (PS).Material and methods. PS-induced liver fibrosis models serve as a primarily immunological mechanism in the development of liver damage and fibrosis.Results. The intraperitoneal administration of THF in rats with PS-induced liver damage produced a reduction of ALT and AST after 60 days. Histopathological changes in liver sections showed an improved histological appearance and lower % of fibrosis after 60 days in liver damaged rats that received THF treatment. Serum IL-6 levels were visibly reduced by THF administration after 60 days and in comparison with rats that did not receive the treatment. This was due to an increment in serum IL-10 levels caused by the administration of THF, which appears to reduce the inflammatory process by decreasing immune response.Conclusion. THF had beneficial effects in combating liver damage and fibrosis processes in an autoimmune model of PS-induced liver fibrosis in rats.

Differential expression of the genes involved in amino acids and nitrogen metabolisms during liver regeneration of mice

Liver regeneration is a highly coordinated response to hepatic injury or resection that is controlled by the body's overall requirement for liver function. The level of circulating amino acids in blood increases after acute liver injury and administration of amino acid mixtures induces hepatic DNA replication. These findings suggest a close connection between amino acid metabolism and hepatic proliferation. However, the underlying molecular mechanisms have not been completely elucidated. Here, we applied a cDNA micro-array technique to analyze expression profiles of the genes associated with nitrogen and amino acid metabolism during liver regeneration in mice following treatment with CCl(4). Seventy-nine genes were identified for their significantly altered expression patterns at different stages of liver damage and regeneration. We observed that the numbers of down-regulated genes were remarkably higher than that of up-regulated genes at 1.5 days following carbon tetrachloride administration when hepatic DNA replication was most active, indicating the existence of a counter balance between cell proliferation and liver metabolism functions. Our results suggest that suppression of amino acids metabolism after acute liver injury results in the accumulation of amino acids in plasma that serves as a driving force for liver regeneration.

Identification and preliminary validation of novel biomarkers of acute hepatic ischaemia/reperfusion injury using dual-platform proteomic/degradomic approaches

Hepatic ischaemia/reperfusion (I/R), a major cause of liver damage associated with multiple trauma, haemorrhagic and septic shock, and liver transplantation, contributes significantly to multiple organ failure. Development of novel sensitive biomarkers that detect early stages of liver damage is vital for effective management and treatment of ischaemic liver injury. By using high-throughput immunoblotting and cation–anion exchange chromatography/reversed-phase liquid chromatography-tandem mass-spectrometry, we identified several hepatic proteins, including argininosuccinate synthase (ASS) and estrogen sulfotransferase (EST-1), which were degraded in the liver and rapidly released into circulation during I/R injury. ASS accumulated in serum within 10 min, reached a steady state at 30 min, and persisted up until 3 h after reperfusion following 30 min of total hepatic ischaemia. EST-1 appeared rapidly in blood and attained maximum within 1 hour followed by a decline at 3 h of reperfusion. No ASS or EST-1 protein was detected in serum of control or sham operated rats. ASS and EST-1 exhibited greater sensitivity and specificity toward I/R liver injury as compared with alanine aminotransferase (ALT), an established marker of hepatocellular necrosis. In contrast, serum ASS and EST-1 were undetectable in rats with chronic alcoholic liver disease, while the levels of ALT protein were significantly increased. In addition, ASS, but not EST-1 or ALT accumulated in blood only 6 h after treatment with hepatotoxic combination of lipopolysaccharide and D-galactosamine. These data demonstrate the utility of ASS and EST-1 as novel sensitive and specific biomarkers of acute liver ischaemic injury for prospective clinical studies.

Morphology of portal hypertension at the early stage of liver damage induced by CCl4: an experimental study with dogs

To explore the morphological basis of portal hypertension happened at the early stage of liver damage. Sixteen mongrel dogs were injected intraperitoneally with carbon tetrachloride (CCl(4)) once a week for 4 weeks and then once every 4 days, totally for 7 weeks. Before the experiment and 7 weeks after the beginning of experiment, small pieces of liver tissues were taken through a median upper abdominal incision and underwent HE staining, Masson's staining, and immunohistochemistry with alpha-smooth muscle actin (alpha-SMA) to label the activated astrocyte (hepatic stellate cell, HSC), and transmission electron microscopy. The portal vein pressure was measured by puncture of gastroepiploic vein. The diameter of spleen was measure too. Since the second weeks after the experiment, the dogs showed vomiting, diarrhea, anorexia, weight loss, and varicose vein in the abdominal wall. One dog died of toxic enteritis 3 weeks after the beginning of experiment. 7 weeks after the injection of CCl(4), the average portal vein pressure of the rest 115 dogs was 22.7 cm H(2)O +/- 1.5 cm H(2)O, significantly higher than that before the experiment (12.2 cm H(2)O +/- 1.9 cm H(2)O, 1cm H(2)O = 0.098 kPa, P < 0.05). The long diameter of spleen of the 15 dogs was 24.4 +/- 3.1 cm, significantly greater than that before the experiment (18.7 cm +/- 2.4 cm, P < 0.05). Seven weeks after the injection of CCl(4) the appearance of liver did not change remarkably among the dogs, however, the spleens of the dogs showed swelling and stagnation of blood. No obvious ascites was seen. HE staining showed slight liver damage, such as fatty degeneration of liver cells; Masson's staining and electron microscopy showed typical sinusoid capillarization. alpha-SMA immunohistochemistry stain showed increased number of activated HSCs. Immunohistochemistry showed a number of alpha-SMA positive cells in the hepatic lobules. Portal hypertension was successfully induced in 15 dogs. At the early stage of liver damage the possible cause of portal hypertension is the increased resistance of blood stream due to sinusoid capillarization and activation of HSC.

Liver biopsy in hepatitis C

The advent of more specific and sensitive assays for the diagnosis of hepatitis C has provided better definition of the histopathology of this disease and its differentiation from other forms of hepatitis. The following features have been found to be characteristic but not pathognomonic: lymphoid follicles in the portal tracts, bile duct lesions, fatty metamorphosis and Mallory body-like condensations in the cytoplasm of the hepatocytes. It has been suggested that the virus genotype may be correlated with different histopathologies. A better understanding of chronic hepatitis C has led to a more flexible and accurate terminology of chronic hepatitis, including the grade of necroinflammatory activity and the stage of fibrosis. Both seem to be of prognostic value in the course of chronic hepatitis C. Recurrence of hepatitis C in the graft after liver transplantation may have a different histopathologic pattern with progressive fibrosis, conspicuous cholestasis and pronounced hepatocellular necrosis. Histopathology grade and stage of liver damage in chronic hepatitis C is not reflected by serum aminotransaminase level or serological status.