Early Stage Of Hepatic Fibrosis Research Articles

Liver stiffness as measured by two-dimensional shear wave elastography overestimates the stage of fibrosis in patients with chronic hepatitis B and hepatic steatosis

Two-dimensional shear wave elastography (2D-SWE) is a non-invasive technique for measuring liver stiffness (LS) and is used to assess the degree of hepatic fibrosis in patients with chronic hepatitis B (CHB). Despite its usefulness, several factors, other than hepatic fibrosis, can affect its diagnostic accuracy. Hepatic steatosis (HS) is a common lesion in CHB that has increasingly been getting attention in the field of disease development; however, its influence on the measurement of LS remains unclear. We aimed to determine whether HS affects the diagnostic accuracy of 2D-SWE in patients with CHB. Serum parameters and LS values were obtained from 161 patients with CHB. The degrees of hepatic fibrosis and inflammatory activity were estimated based on the METAVIR Cooperative Study Group criteria, and the extent of HS was defined as the percentage of hepatocytes containing fat droplets using oil red staining. We found that LS values were independently correlated with HS in the early stages of hepatic fibrosis (F0-F2 or F0-3). Furthermore, LS values in patients with significant steatosis (S≥10%) were higher than the counterpart in fibrosis stages F0-2 (6.82±1.57 vs. 7.92±1.99; p=0.010) and F0-3 (7.18±1.84 vs. 8.25±1.91; p=0.007). Therefore, false positive rates (FPRs) in the diagnosis of advanced fibrosis (16.00% vs. 37.04%, p=0.037) and cirrhosis (6.67% vs. 21.62%, p=0.030) were higher in patients with significant steatosis. In conclusion, the use of 2D-SWE in the measurement of LS overestimates the stage of hepatic fibrosis in CHB patients with HS>10%. This should be taken into consideration to combine LS results with other non-invasive parameters to improve its accuracy.

Expression of Beclin-1, Bcl-2, Bcl-xL, Bad, and Bax in HCV Patients in Relation to Grade of Hepatic Fibrosis

Autophagy plays an important role in the pathogenesis of many diseases. However, its role is still unclear. we investigate the mRNA expression of Beclin-1 (major autophagic agent), pro-apoptotic agents (Bad, Bax), and anti-apoptotic agents (Bcl-2, Bcl-xL) in blood samples withdrawn from Genotype 4 HCV-infected patients with different stages of hepatic fibrosis. The study was a retrospective one that included 30 healthy people (Control Group), 64 chronic hepatitis C patients with early hepatic fibrosis stages [grade 0 and 1 fibrosis] (F0-1 Group), and 36 chronic hepatitis C patients with Late hepatic fibrosis stages [grade 2 and 3 fibrosis] (F2-3 Group). qPCR was used to measure mRNA expression in the samples. Beclin-1, Bad, and Bax mRNA expression in F0-1 Group were significantly higher than both F2-3 Group and Control Group (P<0.001). While Bcl-2, and Bcl-xL mRNA expression in F0-1 Group were significantly lower than both F2-3 Group and Control Group (P<0.001). Beclin-1, Bad, and Bax mRNA expression were increased at the early stages of hepatic fibrosis in HCV patients, and were declined as the fibrosis progressed to more advanced stages, while Bcl-2, and Bcl-xL mRNA expression were increased as fibrosis progresses. This shows that Autophagy has an important role in the early stages of hepatic fibrosis in Genotype 4 HCV patients. These findings provide an insight into the pathogenesis of chronic HCV infection, and the effect of autophagy on liver fibrosis. This may be used to provide possible biomarkers and contribute to a new therapeutic approach.

Machine-learning-based classification of real-time tissue elastography for hepatic fibrosis in patients with chronic hepatitis B

Hepatic fibrosis is a common middle stage of the pathological processes of chronic liver diseases. Clinical intervention during the early stages of hepatic fibrosis can slow the development of liver cirrhosis and reduce the risk of developing liver cancer. Performing a liver biopsy, the gold standard for viral liver disease management, has drawbacks such as invasiveness and a relatively high sampling error rate. Real-time tissue elastography (RTE), one of the most recently developed technologies, might be promising imaging technology because it is both noninvasive and provides accurate assessments of hepatic fibrosis. However, determining the stage of liver fibrosis from RTE images in a clinic is a challenging task. In this study, in contrast to the previous liver fibrosis index (LFI) method, which predicts the stage of diagnosis using RTE images and multiple regression analysis, we employed four classical classifiers (i.e., Support Vector Machine, Naïve Bayes, Random Forest and K-Nearest Neighbor) to build a decision-support system to improve the hepatitis B stage diagnosis performance. Eleven RTE image features were obtained from 513 subjects who underwent liver biopsies in this multicenter collaborative research. The experimental results showed that the adopted classifiers significantly outperformed the LFI method and that the Random Forest(RF) classifier provided the highest average accuracy among the four machine algorithms. This result suggests that sophisticated machine-learning methods can be powerful tools for evaluating the stage of hepatic fibrosis and show promise for clinical applications.

Metabolic characterization of the early stage of hepatic fibrosis in rat using GC-TOF/MS and multivariate data analyses.

The aim of this study was to explore the changes in the urine metabolic spectrum in rats with the early stage of liver fibrosis using gas chromatography-time of flight/mass spectrometry (GC-TOF/MS), try to search for potential biomarkers and elucidate the probably metabonomic pathogenesis. The early stage of liver fibrosis was established with a single subcutaneous injection of carbon tetrachloride twice each week for 4 weeks continuously. At the end of the experiment, GC-TOF/MS technology with multivariate statistical approaches such as principal component analysis, partial least squares-discriminant analysis and orthogonal partial least squares-discriminant analysis was used to analyze the changes in the metabolic spectrum trajectory and identify potential biomarkers. Twelve potential biomarkers in the model group, such as succinic acid, threonine and lactose, were selected, which indicate that the metabonomic pathogenesis of the early stage of liver fibrosis may be related to disorders of energy metabolism, amino acid metabolism and fatty acid metabolism.

Assessing Degree of Hepatic Fibrosis in Patients with Chronic Liver Disease Using Gadoxetic Acid-Enhanced Liver MRI: Preliminary Results

Purpose: To assess different degree of hepatic fibrosis using gadoxetic acid-enhanced MRI in patients with chronic liver disease. Materials and Methods: 31 patients with chronic liver disease and 7 healthy subjects were studied using a 1.5 Tesla MRI. Unenhanced and dynamically enhanced images were acquired using gadoxetic-acid contrast (0.025 mmol/kg). Hepatobiliary phase was obtained with a 20 minute time delay. Liver relative enhancement (LRE) values were calculated sampling 1 cm Regions Of Interest (ROI) per segment. According to Metavir score, patients were divided into different groups: Group A = F0-F1 (absent/mild fibrosis), Group B = F2 (moderate fibrosis) and Group C = F3-F4 (advanced fibrosis). Mann-Whitney test with Bonferroni correction ad hoc was adopted to compare LRE values between classes. Receiver Operating Characteristics (ROC) curves were created for prediction of different degree of fibrosis. Results: When comparing LRE mean values a statistical difference between F0-F1 and F3-F4 groups (p=0.008) was observed. A significant report was also observed between moderate and advanced fibrosis (p=0.02). In the comparison between group A and group B no significant difference for LRE mean values (p=0.93) was observed. For prediction of advanced fibrosis, ROC curve showed an Area Under Curve (AUC) of 0.793, with sensitivity of 94.12% and specificity of 57.89% using a threshold LRE value 1.0031. For prediction of 1.0031. Conclusion: LRE values showed significant difference only between moderate and advanced fibrosis. In patients with chronic liver disease, the assessment of early stages of hepatic fibrosis (mild or moderate fibrosis) is not statistically reliable using gadoxetic acid-enhanced MRI. Keywords: Contrast media, fibrosis, gadolinium ethoxybenzyl DTPA, liver cirrhosis, liver diseases, magnetic resonance imaging.

Hepatic fibrosis and angiogenesis after bile duct ligation are endogenously expressed vasohibin-1 independent

Liver fibrosis is linked to VEGF-induced angiogenesis. Overexpression of exogenous vasohibin-1, a feedback inhibitor of angiogenesis, has been reported to reduce liver fibrosis after bile duct ligation (BDL). To uncover the function of endogenous vasohibin-1, we performed BDL using vasohibin-1-deficient mice and analyzed liver fibrosis, injury, and angiogenesis. Liver fibrosis was induced by 14-days of BDL in both wild-type and vasohibin-1-deficient mice. The liver sections were stained with anti-CD31 to visualize endothelial cells and with Sirius red to observe fibrotic regions. Total RNAs were purified from the livers and expression of collagen I α1 mRNA was measured by quantitative PCR. Plasma ALT activity was determined to assess liver injury. Surprisingly, the same extents of increases were seen in anti-CD31 and Sirius red stainings, collagen I α1 mRNA expressions, hepatic hydroxyproline contents, and ALT activity after 14-days of BDL in both wild-type and vasohibin-1-deficient mice. There was unexpectedly no difference between these mice, suggesting that anti-fibrogenic and angiogenic activities of the endogenous vasohibin-1 might be masked in the normal liver at early stage of hepatic fibrosis in mice.

Magnetic resonance elastography and acoustic radiation force impulse for staging hepatic fibrosis: a meta-analysis

Elastography is a non-invasive method to quantify fibrosis based on tissue mechanical properties. We performed a meta-analysis to assess the diagnostic accuracy of two such techniques: Acoustic Radiation Force Impulse Imaging (ARFI) or Magnetic Resonance Elastography (MRE) for staging hepatic fibrosis. Literature databases were searched until June 2013. Inclusion criteria were evaluation of MRE or ARFI, liver biopsy, and reported sensitivity and specificity. A random effects model was used to combine sensitivity and specificity, from which positive (LR+) and negative (LR-) likelihood ratios, diagnostic odds ratios, and area under receiver operating characteristics curve (AUROC) were derived. Differences between MRE and ARFI were compared with t tests (P < 0.05 considered significant). Eleven MRE studies including 982 patients and fifteen ARFI studies including 2,128 patients were selected. AUROC for MRE staging fibrosis were 0.94, 0.97, 0.96, and 0.97 for F1-F4, respectively, whereas AUROC for ARFI staging were 0.82, 0.85, 0.94, and 0.94 for F1-F4, respectively. Significance was found in AUROC between MRE and ARFI for the diagnosis of stage 1 and 2 fibrosis. MRE is more accurate than ARFI with a higher combination of sensitivity, specificity, LR, and AUROC particularly in diagnosing early stages of hepatic fibrosis.

Expression of angiotensinogen during hepatic fibrogenesis and its effect on hepatic stellate cells

SummaryBackgroundThe liver renin-angiotensin system (RAS) plays an important role in promoting the development of hepatic fibrogenesis. Angiotensinogen (AGT) is an important precursor in tissue RAS. This study aimed to investigate the expression and cellular source of AGT in hepatic fibrogenesis and its effect on proliferation and collagen metabolism of hepatic stellate cells.Material/MethodsIn a rat carbon tetrachloride (CCl4)-induced liver fibrosis model the mRNA expression of AGT was determined by real-time PCR and the cellular source of AGT was determined by immunohistochemical staining. In vitro HSC-T6 cells were transfected with AGT, and the expression plasmid, AGT shRNA plasmid and negative shRNA plasmid were constructed. Real-time PCR and ELISA were applied to determine the mRNA expressions and contents of TIMP-1, TGF-β1, type I collagen and type III collagen of the cells or in the supernatants.ResultsCompared to normal liver, the AGT and α-SMA mRNA expressions increased at the early stage of hepatic fibrosis and decreased in hepatic cirrhosis. The expressions of AGT and α-SMA mRNA were correlated with the hepatic fibrosis (r=0.915, P=0.03). Immunohistochemistry demonstrated the activated HSCs were the main source of AGT due to colocalization of AGT and α-SMA expressions. The mRNA and protein of TGF-β1, TIMP-1, type I collagen and type III collagen were markedly up-regulated.ConclusionsACEI and angiotensin II type 1 receptor antagonist (AT1RA) could attenuate the progression of hepatic fibrosis in the early stage. Direct inhibition of AGT from aHSCs may become an effective antifibrotic anti-liver fibrosis strategy.

Cytokine secretion profile associated with periportal fibrosis in S. mansoni-infected Egyptian patients

Periportal fibrosis (PPF) is a major pathological consequence of S. mansoni infection. Ultrasonography is a well-established tool for diagnosis and grading of schistosomiasis-related pathology. This work is performed to study the effect of schistosomiasis mansoni infection on the cytokine secretion profile in S. mansoni-infected patients at various grades of fibrosis, as determined by ultrasonography using Cairo Working Group classification. The levels of IL-2, IL-4, IL5, IL-10, IFN-gamma, and TNF-alpha were measured in the absence of in vitro antigen stimulation and after stimulation with worm and egg antigens. Simple intestinal (INT) patients are characterized by strong proliferation to worm antigen and high levels of IL-10 and TNF-alpha compared to patients at various grades of infection. GradeII (GdII)-infected patients are characterized by higher IL-5 production than are patients with other clinical forms of the disease. Sharp reduction of almost all cytokines in response to both worm and egg antigens was detected in GdIII-infected patients. These results stressed the role of both IL-10 and TNF-alpha in the early stages of hepatic fibrosis, while IL-5 could be employed as a potential predictive marker for advanced stages. In conclusion, PPF is associated with cytokine production profiles that vary with the magnitude of the fibrosis.

Cytokine profile associated with human chronic schistosomiasis mansoni.

This study objective was to evaluate the cytokines associated with early events of hepatic fibrosis in schistosomiasis mansoni. Hepatic fibrosis was classified by ultrasonography in 94 patients. Immunological evaluation was performed by measurement of secreted cytokines (interleukin IL-5, IL-10, IL-13, interferon-gamma, tumor necrosis factor-alpha and transforming growth factors-beta) in peripherl blood mononuclear cells stimulated by Schistosoma mansoni antigens. Significantly, higher levels of IL-5, IL-10 and IL-13 were found in supernatants of SEA-stimulated PBMC from subjects with degree III hepatic fibrosis as compared to patients with degree I or II fibrosis, Significant increases in IL-5 and IL-13 levels were also observed in some of the subjects who remained untreated for one year following initial assessment and developed more serious fibrosis during this period. The data suggests a role for type 2 cytokines in early stages of hepatic fibrosis in human schistosomiasis mansoni.

Reversibility of hepatic fibrosis: from the first report of collagenase in the liver to the possibility of gene therapy for recovery.

Since the authors reported the presence of collagenase in the liver as well as its increased activity in the early stage of hepatic fibrosis and its reduced activity in advanced fibrosis in rats induced by chronic CCl4 intoxication, in baboons fed alcohol chronically and in patients with alcoholic fibrosis, other investigators have demonstrated the same tendency of collagenase activity biologically and histochemically. Very recently, the authors demonstrated definite gene expression of collagenase during the recovery from experimental hepatic fibrosis using Northern blotting and in situ hybridization. The findings of in situ hybridization not only demonstrated the cells expressing collagenase, but also suggested much information on the mechanism of the recovery from fibrosis. Hepatic stellate cells play a key role not only in fibrogenesis but also in fibrolysis. The authors' recent observation revealed that collagenase (matrix metalloproteinase-13 (MMP-13)) gene expression appears very early in the process of recovery from liver fibrosis, and that both stellate cells and hepatocytes express MMP-13. Recovery from liver cirrhosis requires the gene expression of collagenase, increased production of the collagenase enzyme, and activation of the enzyme balanced with the specific inhibitors of collagenase. The understanding of molecular mechanisms of MMP-1 gene expression which is under investigation in our laboratory may provide us a new strategy for the treatment of liver fibrosis including the possibility of gene therapy.

Molecular mechanism of the reversibility of hepatic fibrosis: with special reference to the role of matrix metalloproteinases.

The participation of matrix metalloproteinases (MMP) and their specific inhibitors, the tissue inhibitors of matrix metalloproteinases (TIMP), in both the formation and degradative recovery processes of liver fibrosis were mainly reviewed from the molecular biological aspect. Since authors first reported increased activity of interstitial collagenase in the early stage of hepatic fibrosis in rats induced by chronic CCl4 intoxication, in baboons fed alcohol chronically and in patients with alcoholic fibrosis, other investigators have also demonstrated increased activity biologically and histochemically. However, species-specific differences in response have been found and gene-level research on the rat model has not demonstrated increased mRNA transcription of collagenase. It has also been clarified that activated stellate cells can also produce matrix components. Very recently, authors observed the participation of interstitial collagenase in the recovery from experimental hepatic fibrosis by using polymerase chain reaction northern blotting and in situ hybridization. The in situ hybridization findings not only demonstrated the cells responsible for interstitial collagenase, but also suggested a great deal about the mechanism of recovery from fibrosis. Hepatic stellate cells are activated via the expression of c-myb and nuclear factor-kappaB (NFkappaB) which is induced by oxidative stress, and inhibited by antioxidant (1-alpha-tocopherol) and butylated hydroxytoluene. The activation mechanism is now being revealed. The relationship between the activation mechanism of stellate cells and the production and secretion of MMP and TIMP in the formation and recovery process of hepatic fibrosis should be investigated from the promoter gene level. This approach might help develop a new strategy for the treatment of liver fibrosis.

A morphological study of the early stages of hepatic fibrosis induced by low doses of dimethylnitrosamine in the rat

Hepatic fibrosis has been induced in rats by low doses of dimethylnitrosamine (DMN) and special attention has been paid to early morphological events. DMN (10 microliter/kg body wt., i.p.) was given 3 days a week for 3 weeks to Sprague-Dawley rats. Liver samples were taken on days 7, 14, 21, 28 and 35 and examined by light and electron microscopy. Hemorrhagic necrosis, mainly centrolobular, was evident on day 7, with disruption of the sinusoidal lining, and widening or disappearance of the spaces of Disse invaded by erythrocytes and lymphocytes. Large granular lymphocytes similar to pit cells were also present in close contact with hepatocytes. At day 14, fibrotic septa were associated with cells bearing 'transitional' features between those of lipocytes, myofibroblasts and fibroblasts. Hepatocytes showed foci of increased smooth endoplasmic reticulum, and altered sinusoidal and canalicular membranes. At day 21, all animals showed nodularity of the parenchyma, with evidence of micronodular cirrhosis associated with ascites in two animals. At day 35 (19 days after cessation of treatment) there was little residual inflammation, but well-defined micronodules were still present in all animals. This shows that, in the rat, 3-week treatment with a low dose of DMN produces micronodular cirrhosis following diffuse hemorrhagic necrosis without steatosis. The response of the animals was uniform and reproducible. Lesions of the sinusoidal wall and of membranes of liver cells associated with the inflammatory reaction appeared prominent.

Drug Metabolizing Activity in Rats with Chronic Liver Injury Induced by Carbon Tetrachloride: Relationship with the Content of Hydroxyproline in the Liver

AbstractThe drug metabolizing activity of rat liver during long-term administration of carbon tetrachloride (CCl4), and its relationship with the content of hydroxyproline (Hyp) in the liver were examined. The contents of cytochrome P-450 (P-450) and cytochrome b5 (b5) and the metabolizalion of aniline, aminopyhne. 7-ethoxycoumarin (7-EC) and benzo(a)pyrene (B(a)P) in the microsomal fraction were examined five days after the final administration of CCl4. The contents of P-450 and b5 and the activity to metabolize the four substrates were gradually reduced as the Hyp content in the liver increased. However, aminopyhne N-demethylation and B(a)P hydroxylation. particularly the latter, was more reduced than aniline hydroxylation and 7-EC O-deethylation in the early stage of hepatic fibrosis. Such differences may be due mainly to the different P-450 subtypes affected by CCl4.