Fibrosis In Chronic Liver Disease Research Articles

A-250 Evaluation of Liver Fibrosis in Schistosomiasis Mansoni Using the ELF (Enhanced Liver Fibrosis) Score

Abstract Background Measuring the presence and pattern of periportal fibrosis (PPF) in schistosomiasis mansoni (SM) is essential for diagnosing its clinical forms and detection of portal hypertension. For these evaluations, the gold-standard has been ultrasound scan (US) using Niamey classification, however it requires sophisticated equipment and well-trained operator. ELF (Enhanced Liver Fibrosis) is an index capable of quantifying liver fibrosis in chronic liver diseases, although there have been few studies evaluating PPF in SM. Aim To assess serum ELF levels according to the PPF patterns established by ultrasound using the Niamey classification in patients with schistosomiasis. Methods A cross-sectional, bidirectional analytical study was carried out with patients from an endemic area (patterns A, B, C and D) and from an SM reference outpatient clinic (patterns D, E and F). Clinical data and 10 mL of blood were collected, centrifuged and frozen to measure and calculate the score by the ELF™ Test (Atellica® IM), according to the equation: [ELF = 2.278 + 0.851 ln(HA) + 0.751 ln(PIIINP) + 0.394 ln(TIMP-1)] and expressed as a numerical value without unit. US using GE Healthcare Logic E (endemic area) or Siemens Acuson S2000 (outpatient clinic) equipment were performed by a single examiner. Results Of the 196 patients evaluated, 114 (58.2%) were women with a mean age of 45 ± 15.42 years. They had the following PPF patterns: 18 pattern A+B; 64 pattern C; 56 pattern D; 49 pattern E and 9 pattern F. The median ELF values were higher according to PPF progression in these 5 groups (p = 0.0021), respectively: (A+B = 8.36); (C = 8.40); (D = 8.64); (E = 8.99) and (F = 9.75). The ELF was able to differentiate patients without PPF, with mild, moderate and advanced PPF: A+B vs C vs D vs E+F (p = 0.0024), as well as patients with advanced PPF from those without fibrosis or with moderate fibrosis: A+B vs E+F (p = 0.0464) and C+D vs E+F (p = 0.0007). The index was also able to differentiate patients with absent or mild PPF from those with moderate or advanced PPF: A+B+C vs D+E+F (p = 0.0015). Conclusions The ELF showed great potential as a tool for assessing PPF in patients with schistosomiasis mansoni, being able to differentiate those with little fibrosis from those with moderate or advanced fibrosis.

KASL clinical practice guidelines for noninvasive tests to assess liver fibrosis in chronic liver disease.

KASL clinical practice guidelines for noninvasive tests to assess liver fibrosis in chronic liver disease.

Letter to the Editor: Imaging-based noninvasive liver disease assessment for staging liver fibrosis in chronic liver disease: A systematic review supporting the AASLD Practice Guideline.

Letter to the Editor: Imaging-based noninvasive liver disease assessment for staging liver fibrosis in chronic liver disease: A systematic review supporting the AASLD Practice Guideline.

Characteristics of liver volume and pathological changes with different stages of liver fibrosis in chronic liver disease

Objective: To measure the overall and lobulated volume of the liver with different degrees of liver fibrosis and to further observe pathological changes such as liver microvasculature, hepatocyte apoptosis, and regeneration in order to understand the macroscopic volume changes of the liver during liver fibrosis and its relationship with liver tissue microscopic pathology in patients with chronic liver disease. Methods: 53 patients with chronic hepatitis B, alcoholic fatty liver disease, autoimmune liver disease, nonalcoholic fatty liver disease, and drug-induced chronic liver disease who underwent both liver biopsy tissue and abdominal magnetic resonance imaging were collected. Patients were divided into early (F1-2), middle (F3-4), and late (F5-6) in accordance with the Ishak fibrosis stage and Masson stain. The liver and spleen volumes were measured using ITK-SNAP software. CD31 immunohistochemical staining was used to reflect intrahepatic angiogenesis. Ki67 and HNF-4α multiplex immunohistochemical staining were used to reflect hepatocyte regeneration. GS staining was used to determine parenchymal extinction lesions. TUNEL staining was used to observe hepatocyte apoptosis. Spearman correlation analysis was used to analyze the relationship between liver volume changes and liver histopathological changes. Results: As liver fibrosis progressed, the total liver volume and right lobe liver volume gradually decreased (P<0.05), while the spleen volume gradually increased (P<0.05). The expression of CD31 and GS gradually increased (P<0.05), and the expression of Ki67 first increased and then decreased (P<0.05). The positivity rate of CD31 was negatively correlated with the right lobe liver volume (r=-0.609, P<0.001) and the total liver volume (r=-0.363, P=0.017). The positivity rate of Ki67 was positively correlated with the right lobe liver volume (r=0.423, P=0.018), while the positivity rate of apoptotic cells was significantly negatively correlated with the total liver volume (r=-0.860, P<0.001). The positivity rate of GS was negatively correlated with the right lobe liver volume (r=-0.440, P=0.002), and the number of PELs was negatively correlated with RV (r=-0.476, P=0.013). The CD31 positive staining area was negatively correlated with the Ki67 positive staining area(r=-0.511, P=0.009). Conclusion: As liver fibrosis progresses, patients with chronic liver disease have a depletion in total liver volume and right lobe liver volume, and this is mainly in correlation with fewer liver cells and liver tissue microvasculature disorders.

Reply: Imaging-based noninvasive liver disease assessment for staging liver fibrosis in chronic liver disease: A systematic review supporting the AASLD Practice Guideline.

Reply: Imaging-based noninvasive liver disease assessment for staging liver fibrosis in chronic liver disease: A systematic review supporting the AASLD Practice Guideline.

Enhanced liver fibrosis score is stable after withdrawal in patients with heavy alcohol consumption: A pilot study.

Enhanced liver fibrosis (ELF) score is an accurate, noninvasive test for assessing the severity of liver fibrosis in chronic liver disease, including alcohol-related liver disease. However, whether the ELF score changes during alcohol withdrawal is unknown. This pilot study assessed changes in the ELF score during withdrawal in patients with a history of excessive alcohol intake. In this prospective study, ELF was performed on day 0 (D0, at the beginning of hospitalization), at day 7 (D7, on discharge from hospital), and at follow-up visits on days 30 (D30) and 90 (D90). Transient elastography (TE) was also assessed on days 4 (D4) and D30. The study included 35 patients (71% male) with a mean alcohol intake of 139 g/day. On D30 and D90, 8 and 13 patients had resumed alcohol consumption (mean intake of 90 and 80 g/day, respectively). In patients who remained abstinent, the mean ELF score was 8.93 on D0, 9.14 on D30 (p = 0.32), and 9.27 on D90 (p = 0.14). In patients who resumed alcohol, mean ELF score was 9.7 on D0, 10.05 on D30 (p = 0.09), and 9.71 on D90 (p = 0.12). ELF score was comparable over the first months after withdrawal, although there was a slight increase in the first week (mean ELF score increased from 9.24 on D0 to 9.74 on D7, p < 0.001). Mean TE value was 7.9 kPa on D4 and 8.1 kPa on D30 (p = 0.84) in patients who resumed alcohol consumption, and 8.3 and 7.5 kPa (p = 0.03) on D4 and D30, respectively, in abstinent patients. The ELF score is stable during the first months after withdrawal and thus appears to be a useful tool to assess liver fibrosis or cirrhosis in this setting. Nevertheless, because in the first week there is a transient increase in ELF score, caution in interpretation is warranted.

Progress and Prospects of Elastography Techniques in the evaluation of fibrosis in chronic liver disease

The precise assessment of liver fibrosis degree is imperative for monitoring and managing chronic liver diseases. Traditionally, liver biopsy and quantitative blood biomarkers have been the mainstays for fibrosis assessment, albeit each with its inherent limitations. However, the emergence of elastography presents a promising non-invasive alternative, potentially supplanting or complementing traditional biomarkers. In contrast to conventional biomarkers, elastography offers several advantages. Firstly, it is non-invasive, sidestepping the risks and discomfort associated with liver biopsy. Secondly, this technique provides real-time and intuitive fibrosis assessments, characterized by user-friendly operation and high reproducibility. Thirdly, elastography excels in diagnosing moderate to severe fibrosis, vital for determining treatment strategies and monitoring disease progression. Notably, MRE stands out as the most accurate non-invasive method for fibrosis assessment, especially suitable for advanced fibrotic stages. This article provides an overview of the advancements in elastography as a viable quantitative biomarker for chronic liver disease fibrosis burden.

The Utility of a Novel Stacked Microvascular Imaging for Enhanced Detection of Fibrosis in Chronic Liver Diseases

Ultrasonographic imaging plays a primary role to detect fibrotic changes in patients with chronic liver disease (CLD). To enhance detectability of fibrosis in its early stage, we developed a novel stacked microvascular imaging (SMVI) that enables continuous visualization of fibrotic changes in intrahepatic vessels. SMVI was produced by accumulating 3-5 seconds of high-definition color images in tilted-scan mode. An SMVI score was devised by quantitating three hallmark vascular changes in liver fibrosis in 0-2 grades (total 0-6): narrowing, caliber irregularity, and tortuosity. To evaluate the clinical utility of the SMVI score, 469 well-defined CLD patients were enrolled and subgrouped by the stage of liver fibrosis defined based on elastography: F0-1Low, F0-1High, F2, F3, and F4. The diagnostic performance of the SMVI score was compared to conventional B-mode liver morphology score and various laboratory test markers of fibrosis. Unlike conventional microvascular imaging that relies on a single image, SMVI enabled an undisrupted view of intrahepatic vessels for easy detection of fibrotic changes. SMVI detected microvascular narrowing in 92% at stage F0-1High. While detection rates for caliber irregularity and tortuosity were low at early stages but increased proportionately in advanced stages. Multiple logistic regression analysis revealed that SMVI score was most accurate in distinguishing F0-1Low from F0-1High cases compared to B-mode or laboratory test scores. SMVI provides enhanced vascular images of liver fibrosis in CLD, especially in its early stage. The SMVI score can be used as a primary tool for determining fibrotic stages in CLD.

Economic evaluation of non-invasive liver tests for the diagnosis of liver fibrosis in chronic liver diseases: a systematic review protocol.

The objective of this review is to determine the costs and benefits of non-invasive liver tests vs liver biopsy in patients with chronic liver diseases. Hepatic diseases can lead to liver fibrosis, cirrhosis, and hepatocellular carcinoma. In the past, liver biopsy was the only option for diagnosing fibrosis degree. Liver biopsy is an invasive procedure that depends on the sample size to be able to deliver an accurate diagnosis. In recent years, non-invasive liver tests have been increasingly used to estimate liver fibrosis degree; however, there is a lack of economic assessments of technology implementation outcomes. This review will include partial (cost studies) and complete economic evaluation studies on hepatitis B, hepatitis C, alcoholic liver disease, and non-alcoholic fatty liver disease that compare non-invasive liver tests with liver biopsies. Studies published in English, French, Spanish, German, Italian, or Portuguese will be included. No date limits will be applied to the search. This review will identify published and unpublished studies. Published studies will be identified using MEDLINE (PubMed), Cochrane Library (CENTRAL), Embase, Web of Science, Scopus, and LILACS. Sources of unpublished studies and gray literature will include sources from health technology assessment agencies, clinical practice guidelines, regulatory approvals, advisories and warnings, and clinical trial registries, as well as Google Scholar. Two independent reviewers will screen and assess studies, and extract and critically appraise the data. Data extracted from the included studies will be analyzed and summarized to address the review objective using narrative text, and the JBI dominance ranking matrix. PROSPERO CRD42023404278.

Imaging-based noninvasive liver disease assessment for staging liver fibrosis in chronic liver disease: A systematic review supporting the AASLD Practice Guideline.

Transient elastography (TE), shear wave elastography, and/or magnetic resonance elastography (MRE), each providing liver stiffness measurement (LSM), are the most studied imaging-based noninvasive liver disease assessment (NILDA) techniques. To support the American Association for the Study of Liver Diseases guidelines on NILDA, we summarized the evidence on the accuracy of these LSM methods to stage liver fibrosis (F). A comprehensive search for studies assessing LSM by TE, shear wave elastography, or MRE for the identification of significant fibrosis (F2-4), advanced fibrosis (F3-4), or cirrhosis (F4), using histopathology as the standard of reference by liver disease etiology in adults or children from inception to April 2022 was performed. We excluded studies with <50 patients with a single disease entity and mixed liver disease etiologies (with the exception of HCV/HIV coinfection). Out of 9447 studies, 240 with 61,193 patients were included in this systematic review. In adults, sensitivities for the identification of F2-4 ranged from 51% to 95%, for F3-4 from 70% to 100%, and for F4 from 60% to 100% across all techniques/diseases, whereas specificities ranged from 36% to 100%, 74% to 100%, and 67% to 99%, respectively. The largest body of evidence available was for TE; MRE appeared to be the most accurate method. Imaging-based NILDA outperformed blood-based NILDA in most comparisons, particularly for the identification of F3-4/F4. In the pediatric population, imaging-based NILDA is likely as accurate as in adults. LSM from TE, shear wave elastography, and MRE shows acceptable to outstanding accuracy for the detection of liver fibrosis across various liver disease etiologies. Accuracy increased from F2-4 to F3-4 and was the highest for F4. Further research is needed to better standardize the use of imaging-based NILDA, particularly in pediatric liver diseases.

Accuracy of blood-based biomarkers for staging liver fibrosis in chronic liver disease: A systematic review supporting the AASLD Practice Guideline.

Blood-based biomarkers have been proposed as an alternative to liver biopsy for noninvasive liver disease assessment in chronic liver disease. Our aims for this systematic review were to evaluate the diagnostic utility of selected blood-based tests either alone, or in combination, for identifying significant fibrosis (F2-4), advanced fibrosis (F3-4), and cirrhosis (F4), as compared to biopsy in chronic liver disease. We included a comprehensive search of databases including Ovid MEDLINE(R), EMBASE, Cochrane Database, and Scopus through to April 2022. Two independent reviewers selected 286 studies with 103,162 patients. The most frequently identified studies included the simple aspartate aminotransferase-to-platelet ratio index and fibrosis (FIB)-4 markers (with low-to-moderate risk of bias) in HBV and HCV, HIV-HCV/HBV coinfection, and NAFLD. Positive (LR+) and negative (LR-) likelihood ratios across direct and indirect biomarker tests for HCV and HBV for F2-4, F3-4, or F4 were 1.66-6.25 and 0.23-0.80, 1.89-5.24 and 0.12-0.64, and 1.32-7.15 and 0.15-0.86, respectively; LR+ and LR- for NAFLD F2-4, F3-4, and F4 were 2.65-3.37 and 0.37-0.39, 2.25-6.76 and 0.07-0.87, and 3.90 and 0.15, respectively. Overall, the proportional odds ratio indicated FIB-4 <1.45 was better than aspartate aminotransferase-to-platelet ratio index <0.5 for F2-4. FIB-4 >3.25 was also better than aspartate aminotransferase-to-platelet ratio index >1.5 for F3-4 and F4. There was limited data for combined tests. Blood-based biomarkers are associated with small-to-moderate change in pretest probability for diagnosing F2-4, F3-4, and F4 in viral hepatitis, HIV-HCV coinfection, and NAFLD, with limited comparative or combination studies for other chronic liver diseases.

Cirrhotic cardiomyopathy: a subject that's always topical.

Cirrhosis is the final stage in the development of hepatic fibrosis in chronic liver disease. It is associated with major hemodynamic disturbances defining the hyperdynamic circulation and may be complicated by specific cardiac involvement or cirrhotic cardiomyopathy which is a silent clinical condition that typically remains asymptomatic until the late stages of liver disease. Recently, new criteria defining CC, based on modern concepts and knowledge of heart failure, have been proposed. Despite knowledge of the main mechanisms behind this entity, there is no specific treatment available for cirrhotic cardiomyopathy. The management approach for symptomatic cardiomyopathy in cirrhotic patients is similar to that for left ventricular failure in non-cirrhotic individuals.

Diagnostic performance of shear wave measurement in the detection of hepatic fibrosis: A multicenter prospective study.

This study aimed to establish the shear wave measurement (SWM) cut-off value for each fibrosis stage using magnetic resonance (MR) elastography values as a reference standard. We prospectively analyzed 594 patients with chronic liver disease who underwent SWM and MR elastography. Correlation coefficients (were analyzed, and the diagnostic value was evaluated by the area under the receiver operating characteristic curve. Liver stiffness was categorized by MR elastography as F0 (<2.61kPa), F1 (≥2.61kPa, <2.97kPa, any fibrosis), F2 (≥2.97kPa, <3.62kPa, significant fibrosis), F3 (≥3.62kPa, <4.62kPa, advanced fibrosis), or F4 (≥4.62kPa, cirrhosis). The median SWM values increased significantly with increasing fibrosis stage (p<0.001). The correlation coefficient between SWM and MR elastography values was 0.793 (95% confidence interval 0.761-0.821). The correlation coefficients between SWM and MR elastography values significantly decreased with increasing body mass index and skin-capsular distance; skin-capsular distance values were associated with significant differences in sensitivity, specificity, accuracy, or positive predictive value, whereas body mass index values were not. The best cut-off values for any fibrosis, significant fibrosis, advanced fibrosis, and cirrhosis were 6.18, 7.09, 8.05, and 10.89kPa, respectively. This multicenter study in a large number of patients established SWM cut-off values for different degrees of fibrosis in chronic liver diseases using MR elastography as a reference standard. It is expected that these cut-off values will be applied to liver diseases in the future.

The Diagnostic Performance of Serum Hyaluronic Acid and Collagen III in Differentiating Liver Fibrosis in HCV-Induced Patients

Abstract Introduction/Objective Chronic liver diseases may lead to inflammation and progressive scarring, causing irreversible hepatic damage (cirrhosis). To avoid the complications of liver puncture biopsy, Collagen III and Hyaluronic acid are studied as non-invasive markers of liver fibrosis in chronic liver diseases. Aim of the work: Discovering the diagnostic role of serum Hyaluronic acid and Collagen III in liver fibrosis associated with chronic HCV patients. Methods/Case Report The study was performed on 118 patients with liver fibrosis associated with chronic HCV (60 males and 58 females) and 50 age-matched healthy subjects (27 males and 23 females). Liver function tests were measured, including ALT, AST, [AST/ ALT] ratio, albumin, total bilirubin, and platelets counts. APRI was also calculated as [AST/ (upper limits of normal)/ platelets count 109/L]×100. Serum levels of Collagen III (ng/ml) and hyaluronic acid (µg/L) were determined. Plotted receiver-operating characteristic (ROC) curves were used to assess and compare the diagnostic accuracy of biomarkers. Results (if a Case Study enter NA) The area under ROC curve of collagen III in discriminating liver fibrosis in patients associated with chronic HCV infection and healthy ones was 0.805; P&amp;lt; 0.0001. The Sensitivity and Specificity were 62% and 96%, respectively. While the area under ROC curve of hyaluronic acid in discriminating liver fibrosis patients associated with chronic HCV infection from healthy volunteers was 0.832; P &amp;lt; 0.0001. The Sensitivity and Specificity were 72% and 94%, respectively. Also, there was no significant correlation between levels of collagen III and hyaluronic acid; (r=0.01 &amp; P=0.714). Conclusion Collagen III and Hyaluronic acid can identify patients with liver fibrosis with a detection rate of 62% and 72%, respectively. So, both biomarkers are considered powerful non-invasive methods in discriminating and diagnosing liver fibrosis in chronic HCV patients.

Gut Microbiota and Bacterial Translocation in the Pathogenesis of Liver Fibrosis.

Cirrhosis is the end result of liver fibrosis in chronic liver diseases. Studying the mechanisms of its development and developing measures to slow down and regress it based on this knowledge seem to be important tasks for medicine. Currently, disorders of the gut-liver axis have great importance in the pathogenesis of cirrhosis. However, gut dysbiosis, which manifests as increased proportions in the gut microbiota of Bacilli and Proteobacteria that are capable of bacterial translocation and a decreased proportion of Clostridia that strengthen the intestinal barrier, occurs even at the pre-cirrhotic stage of chronic liver disease. This leads to the development of bacterial translocation, a process by which those microbes enter the blood of the portal vein and then the liver tissue, where they activate Kupffer cells through Toll-like receptor 4. In response, the Kupffer cells produce profibrogenic cytokines, which activate hepatic stellate cells, stimulating their transformation into myofibroblasts that produce collagen and other elements of the extracellular matrix. Blocking bacterial translocation with antibiotics, probiotics, synbiotics, and other methods could slow down the progression of liver fibrosis. This was shown in a number of animal models but requires further verification in long-term randomized controlled trials with humans.

Magnetic susceptibility and R2*-based texture analysis for evaluating liver fibrosis in chronic liver disease

PurposeTo explore potential feasibility of texture features in magnetic susceptibility and R2* maps for evaluating liver fibrosis. MethodsThirty-one patients (median age 46 years; 22 male) with chronic liver disease were prospectively recruited and underwent magnetic resonance imaging (MRI), blood tests, and liver biopsy. Susceptibility and R2* maps were obtained using a 3-dimensional volumetric interpolated breath-hold examination sequence with a 3T MRI scanner. Texture features, including histogram, gray-level co-occurrence matrix (GLCM), gray-level dependence matrix (GLDM), gray-level run length matrix (GLRLM), gray-level size zone matrix (GLSZM), and neighboring gray tone difference matrix (NGTDM) features, were extracted. Texture features and blood test results of non-significant (Ishak-F < 3) and significant fibrosis patients (Ishak-F ≥ 3) were compared, and correlations with Ishak-F stages were analyzed. Areas under the curve (AUCs) were calculated to determine the efficacy for evaluating liver fibrosis. ResultsNine texture features of susceptibility maps and 19 features of R2* maps were significantly different between non-significant and significant fibrosis groups (all P < 0.05). Large dependence high gray-level emphasis (LDHGLE) of GLDM and long run high gray-level emphasis (LRHGLE) of GLRLM in R2* maps showed significantly negative and good correlations with Ishak-F stages (r = –0.616, P < 0.001; r = –0.637, P < 0.001). Busyness (NGTDM) in susceptibility maps, LDHGLE of GLDM and LRHGLE of GLRLM in R2* maps yield the highest AUCs (AUC = 0.786, P = 0.007; AUC = 0.807, P = 0.004; AUC = 0.819, P = 0.003). ConclusionTexture characteristics of susceptibility and R2* maps revealed possible staging values for liver fibrosis. Susceptibility and R2*-based texture analysis may be a useful and noninvasive method for staging liver fibrosis.

Immunohistochemical expression of NRF2 is correlated with the magnitude of inflammation and fibrosis in chronic liver disease.

The nuclear factor E2-related factor 2-Kelch-like Ech-associated protein (NRF2-KEAP1) pathway is a major cellular defense mechanism against oxidative stress. However, the role of NRF2-KEAP1 signaling in the development of chronic liver disease remains unclear. Clinical liver specimens from 50 hepatocellular carcinoma (HCC) developed from non-alcoholic steatohepatitis (NASH), 49 HCCs developed from chronic viral hepatitis C (CHc), and 48 liver metastases of colorectal cancer (CRC) from both tumorous and non-tumorous areas were collected during hepatic resection surgery. They were evaluated by immunohistochemical analyses of hematoxylin-eosin, Masson's trichrome, NRF2, and KEAP1, and compared with clinicopathological information. Hepatic inflammation and fibrosis were more severe in the low-intensity NRF2 group than in the high-intensity NRF2 group both between CRC and NASH (Low vs. High: inflammation; p = 0.003, fibrosis; p = 0.014), and between CRC and CHc (Low vs. High: inflammation; p = 0.031, fibrosis; p = 0.011), which could indicate that NRF2 expression in cytosol of hepatocytes was inversely correlated with liver inflammation and fibrosis in non-tumorous areas. The dense staining of NRF2 in the nuclei of non-tumor hepatocytes positively correlated with liver inflammation (CRC and NASH; R = 0.451, p < 0.001, CRC and CHc; R = 0.502, p < 0.001) and fibrosis (CRC and NASH; R = 0.566, p < 0.001, CRC and CHc; R = 0.548, p < 0.001) in both NASH and CHc, and was inversely correlated with hepatic spare ability features such as platelet count (R = -0.253, p = 0.002) and prothrombin time (R = -0.206, p = 0.012). However, KEAP1 expression was not correlated with NRF2 expression levels and nuclear staining intensity. Nuclear translocation of NRF2 was correlated with the magnitude of liver inflammation and fibrosis in chronic liver disease. These results suggest that NRF2 plays a protective role in the development of chronic liver diseases such as NASH and CHc.

Human liver stem cell-derived extracellular vesicles modulate long non-coding RNA expression profile in an in vivo model of non-alcoholic steatohepatitis

Aim: Modifications in long non-coding RNA (lncRNA) expression are associated with inflammation and fibrosis in chronic liver diseases. It has been recently demonstrated that human liver stem cells (HLSCs) and their extracellular vesicles (EVs) can effectively reduce inflammation and fibrosis in a murine model of non-alcoholic steatohepatitis (NASH). Now it has been evaluated whether EVs can modify the expression of inflammation-related lncRNAs in NASH liver. Methods: To induce NASH, severe combined immunodeficient mice were fed with a methionine-choline-deficient diet for 4 weeks. After 2 weeks of diet, 2.5 × 109 EVs were intravenously injected twice a week. An array of 84 inflammation-related lncRNAs was performed on the RNA isolated from NASH livers, and the expression of 14 selected lncRNAs was then validated by real-time polymerase chain reaction (PCR) analysis. Expression levels of maternally expressed gene 3 (Meg3) were further evaluated in vitro, in an activated human hepatic immortalized stellate cell line (LX-2) stimulated with EVs. Results: The screening showed an altered lncRNA expression profile in the liver of NASH mice, in respect to control healthy mice. EV treatment modulated several inflammation-related lncRNAs in NASH livers. Real-time PCR validation of array results indicated that EVs restored to normal levels the expression of 10 lncRNAs altered in NASH. In particular, EV stimulation reduced Meg3 expression levels, which were increased in NASH as well as in activated LX-2. Conclusions: HLSC-EVs regulate the expression of inflammation-related lncRNAs impaired in NASH livers and in an in vitro model of liver fibrosis.

Artesunate Induces Ferroptosis in Hepatic Stellate Cells and Alleviates Liver Fibrosis via the ROCK1/ATF3 Axis.

Development of fibrosis in chronic liver disease requires activation of hepatic stellate cells (HSCs) and leads to a poor outcome. Artesunate (Art) is an ester derivative of artemisinin that can induce ferroptosis in HSCs, and activated transcriptional factor 3 (ATF3) is an ATF/CREB transcription factor that is induced in response to stress. In this study, we examined the role of the Rho-associated protein kinase 1 (ROCK1)/ATF3 axis in Art-induced ferroptosis in HSCs. HSC activation and ferroptosis were studied in vitro by western blotting, polymerase chain reaction, immunofluorescence, and other assays. ATF3 electrophoretic mobility and ROCK1 protein stability were assayed by western blotting. Immunoprecipitation was used to detect the interaction of ROCK1 and ATF3, as well as ATF3 phosphorylation. A ubiquitination assay was used to verify ROCK1 degradation. Atf3-interfering and Rock1-overexpressing mice were constructed to validate the anti-hepatic fibrosis activity of Art in vivo. Art induced ferroptosis in HSCs following glutathione-dependent antioxidant system inactivation resulting from nuclear accumulation of unphosphorylated ATF3 mediated by ROCK1-ubiquitination in vitro. Art also decreased carbon tetrachloride-induced liver fibrosis in mice, which was reversed by interfering with Atf3 or overexpressing Rock1. The ROCK1/ATF3 axis was involved in liver fibrosis and regulation of ferroptosis, which provides an experimental basis for further study of Art for the treatment of liver fibrosis.

The role of sound touch elastography in assessment of liver fibrosis in chronic liver disease keeping APRI as the reference standard.

The study aims to determine the role of Sound Touch Elastography [STE] technique in staging liver fibrosis and predicting clinically significant gastro-esophageal varices among patients with chronic liver disease [CLD] keeping aspartate aminotransferase to platelet ratio index [APRI] as the reference standard. A prospective short-term study including 60 eligible patients with CLD were staged as non-significant fibrosis [NSF], significant fibrosis [SF] and cirrhosis [C] based on APRI values. STE was performed on each patient obtaining multiple readings as per pre-defined standards. The intra-observer reliability between each measurement and its association with APRI staging was evaluated using relevant statistical variables. Further, Youden's index was used to define the optimum cut-off values on STE in differentiating the stages of fibrosis and in predicting clinically significant gastro-esophageal varices. Based on APRI cut-off values, 41.7% [n=25] of the study population had cirrhosis, while 45% [n=27] had significant fibrosis and 13.3% [n=8] had NSF. The STE values in kPa showed a positive correlation with APRI values [(rs)=0.837, p<0.001]. The intra-class correlation estimates based on a mean rating [k=5] was found to be 0.97 [0.95-0.99], implying an excellent agreement between the measurements. Optimum cut-off values in staging SF and C were 7.26 kPa [J=0.73, sensitivity-85.19%, specificity-87.5%; 95% CI] and 13.79 kPa [J=0.84, sensitivity-96.0%, specificity-88.89%; 95% CI]. The AUROC for each of these stages were 0.926 [0.785-0.987] and 0.976 [0.890-0.999], respectively. 23.3% [n=14] of the study population had clinically significant gastro-esophageal varices with a value above 18.84 kPa [J=0.88] showing a sensitivity of 92.85% and a specificity of 95.65% in predicting the same. The novel STE technique shows good accuracy in staging liver fibrosis as determined by APRI values and in prediction of clinically significant gastro-esophageal varices with excellent reliability. It shows promising prospects and can be integrated widely in clinical practice for assessment and staging of fibrosis in CLD.