Fibrosis In Non-alcoholic Steatohepatitis Patients Research Articles

Collagen-targeted protein nanomicelles for the imaging of non-alcoholic steatohepatitis

In vivo molecular imaging tools hold immense potential to drive transformative breakthroughs by enabling researchers to visualize cellular and molecular interactions in real-time and/or at high resolution. These advancements will facilitate a deeper understanding of fundamental biological processes and their dysregulation in disease states. Here, we develop and characterize a self-assembling protein nanomicelle called collagen type I binding – thermoresponsive assembled protein (Col1-TRAP) that binds tightly to type I collagen in vitro with nanomolar affinity. For ex vivo visualization, Col1-TRAP is labeled with a near-infrared fluorescent dye (NIR-Col1-TRAP). Both Col1-TRAP and NIR-Col1-TRAP display approximately a 3.8-fold greater binding to type I collagen compared to TRAP when measured by surface plasmon resonance (SPR). We present a proof-of-concept study using NIR-Col1-TRAP to detect fibrotic type I collagen deposition ex vivo in the livers of mice with non-alcoholic steatohepatitis (NASH). We show that NIR-Col1-TRAP demonstrates significantly decreased plasma recirculation time as well as increased liver accumulation in the NASH mice compared to mice without disease over 4 hours. As a result, NIR-Col1-TRAP shows potential as an imaging probe for NASH with in vivo targeting performance after injection in mice. Statement of significanceDirect molecular imaging of fibrosis in NASH patients enables the diagnosis and monitoring of disease progression with greater specificity and resolution than do elastography-based methods or blood tests. In addition, protein-based imaging probes are more advantageous than alternatives due to their biodegradability and scalable biosynthesis. With the aid of computational modeling, we have designed a self-assembled protein micelle that binds to fibrillar and monomeric collagen in vitro. After the protein was labeled with near-infrared fluorescent dye, we injected the compound into mice fed on a NASH diet. NIR-Col1-TRAP clears from the serum faster in these mice compared to control mice, and accumulates significantly more in fibrotic livers.This work advances the development of targeted protein probes for in vivo fibrosis imaging.

WED-436 - Validation and utility of artificial intelligence-based zonal annotations as an additional assessment tool for the histopathologic review of fibrosis in non-alcoholic steatohepatitis patients

WED-436 - Validation and utility of artificial intelligence-based zonal annotations as an additional assessment tool for the histopathologic review of fibrosis in non-alcoholic steatohepatitis patients

Correlation of serum markers with FibroScan® liver for assessment of hepatic fibrosis in patients with non-alcoholic steatohepatitis

Abstract Background: Newer non-invasive modalities such as FibroScan and laboratory testing methods such as aspartate aminotransferase (AST) platelet ratio index (APRI) scores, AST/alanine aminotransferase (ALT) ratio and fibrosis-4 (FIB-4) scores are being used for fibrosis assessment in non-alcoholic fatty liver disease (NAFLD) patients where facilities for liver biopsy are lacking. We studied the relationship between FibroScan® scores and serum fibrosis markers in non-alcoholic steatohepatitis (NASH) patients. Methods: One hundred patients who had undergone FibroScan of the liver over the past 2 years were enrolled. Demographic data were collected along with serum ALT, serum AST and platelet counts. The AST/ALT ratio, APRI score and FIB-4 were compared with the FibroScan fibrosis scores. Correlate between the degree of fibrosis in relation to age, platelet counts, serum ALT, serum markers and FibroScan scores was studied. Results: A significant positive correlation was noted between the FibroScan® results and the AST/ALT ratio, the APRI score and the FIB-4. No significant correlation was noted between age and fibrosis score. There was a highly significant correlation between platelet count and stiffness score. Conclusions: Our observations suggest that FibroScan®, along with the serum fibrosis markers, is helpful in assessing liver fibrosis in NASH patients, eliminating the need for liver biopsy in resource-poor settings.

FRI068 - Impact of artificial intelligence assistive tool on the histopathologic review of fibrosis in non-alcoholic steatohepatitis patients

FRI068 - Impact of artificial intelligence assistive tool on the histopathologic review of fibrosis in non-alcoholic steatohepatitis patients

Noninvasive biomarkers in predicting nonalcoholic steatohepatitis and assessing liver fibrosis: systematic review and meta-analysis.

Nonalcoholic steatohepatitis (NASH) is characterized by hepatic steatosis with inflammation, ballooned hepatocytes and possible fibrosis, which may progress to liver cirrhosis. Although liver biopsy, remains the diagnostic gold standard of NASH, several noninvasive biomarkers have been studied, to avoid the need for this invasive procedure. We performed a systematic review with meta-analysis to evaluate the accuracy of several noninvasive biomarkers in predicting NASH and assessing liver fibrosis in NASH patients. An electronic search on PubMed and Embase was systematically performed. The principal summary outcome was the area under the curve (AUC), assessing the accuracy of NashTest, BARD (Body Mass Index, AST/ALT ratio, diabetes) score, NAFLD fibrosis score (NFS), APRI (aspartate aminotransferase-to-Platelet Ratio Index), and Fibrosis-4 (FIB-4) Index in predicting NASH and assessing liver fibrosis. Thirteen studies involving 6557 adult patients were included in the qualitative assessment of this review, out of which, six studies were included in the quantitative assessment. Prediction of NASH was evaluated better using NFS (AUC of 0.687) and FIB-4 (AUC of 0.729). Fibrosis stages 0 vs. 1-4 was diagnosed better using NFS (AUC of 0.718) and FIB-4 (AUC of 0.723). Advanced fibrosis was assessed better by BARD (AUC of 0.673), APRI (AUC of 0.762), NFS (AUC of 0.787) and FIB-4 (AUC of 0.821). FIB-4 predicted NASH and quantified liver fibrosis, stages 0 vs. 1-4 more precisely compared to NFS, APRI, and BARD. However, considering that methodological quality of the assessed studies is limited, the results should be considered with caution.

Serum Wisteria floribunda agglutinin-positive Mac-2-binding protein expression predicts disease severity in nonalcoholic steatohepatitis patients.

The role of Wisteria floribunda agglutinin-positive Mac-2 binding protein (WFA+ -M2BP) in the prediction of disease severity in nonalcoholic fatty liver disease (NAFLD) remains elusive. This study evaluated the performance of WFA+ -M2BP in predicting fibrosis in patients with NAFLD. A total of 80 patients with biopsy-proven nonalcoholic steatohepatitis (NASH) were enrolled. Serum WFA+ -M2BP levels were measured using standard methods. The fibrosis-4 (FIB-4) index was also measured. The mean values of WFA+ -M2BP were 1.0, 1.0, 0.8, and 2.2 in Metavir fibrosis stage F0, F1, F2, and F3-4, respectively (linear trend p= 0.005). The optimal cut-off value of WFA+ -M2BP in predicting advanced fibrosis (F3-4) was 1.37 cut-off index (COI), yielding the sensitivity, specificity, positive predictive value (PPV), negative predictive value, and accuracy of 75.0, 79.4, 39.1, 94.7, and 78.7%, respectively (p< 0.001). Combining WFA+ -M2BP with FIB-4 significantly increased the diagnostic performance for advanced fibrosis, yielding specificity, PPV, and accuracy of 100, 100, and 93%, respectively. The significant factors predicting advanced liver fibrosis in the multivariate regression analysis were WFA+ -M2BP ≥ 1.37 COI (OR/confidence interval [CI]: 9.49/1.63-55.21, p= 0.01) and FIB-4≥ 2.80 (OR/CI: 38.18/4.89-297.93, p= 0.001). Monitoring WFA+ -M2BP is suitable for noninvasive assessment of liver fibrosis in NASH patients, particularly in combination with FIB-4.

Obeticholic acid (OCA) improves non-invasive markers of fibrosis in patients with non-alcoholic steatohepatitis (NASH): a secondary analysis of the phase 3 Regenerate study

REGENERATE showed that treatment with OCA improved histological liver fibrosis in NASH patients.1 However, liver biopsy is an impractical tool to monitor patients’ response to therapy. Here, we evaluate the potential utility of non-invasive tests in monitoring NASH patients with fibrosis during treatment. Patients with fibrosis stages 2 and 3 were randomized (1:1:1) to placebo, OCA 10mg or OCA 25mg QD. Change in non-invasive biomarker scores of fibrosis (FIB-4 index, AST to platelet ratio index [APRI]), FibroSure® and NASH (CK-18), and liver stiffness via transient elastography (TE, subset of patients), were assessed.

AEBP1 expression increases with severity of fibrosis in NASH and is regulated by glucose, palmitate, and miR-372-3p.

Factors governing the development of liver fibrosis in nonalcoholic steatohepatitis (NASH) are only partially understood. We recently identified adipocyte enhancer binding protein 1 (AEBP1) as a member of a core set of dysregulated fibrosis-specific genes in human NASH. Here we sought to investigate the relationship between AEBP1 and hepatic fibrosis. We confirmed that hepatic AEBP1 expression is elevated in fibrosis compared to lobular inflammation, steatosis, and normal liver, and increases with worsening fibrosis in NASH patients. AEBP1 expression was upregulated 5.8-fold in activated hepatic stellate cells and downregulated during chemical and contact induction of biological quiescence. In LX-2 and HepG2 cells treated with high glucose (25 mM), AEBP1 expression increased over 7-fold compared to normal glucose conditions. In response to treatment with either fructose or palmitate, AEBP1 expression in primary human hepatocytes increased 2.4-fold or 9.6-fold, but was upregulated 55.8-fold in the presence of fructose and palmitate together. AEBP1 knockdown resulted in decreased expression of nine genes previously identified to be part of a predicted AEBP1-associated NASH co-regulatory network and confirmed to be upregulated in fibrotic tissue. We identified binding sites for two miRNAs known to be downregulated in NASH fibrosis, miR-372-3p and miR-373-3p in the AEBP1 3’ untranslated region. Both miRNAs functionally interacted with AEBP1 to regulate its expression. These findings indicate a novel AEBP1-mediated pathway in the pathogenesis of hepatic fibrosis in NASH.

Validation of PNPLA3 polymorphisms as risk factor for NAFLD and liver fibrosis in an admixed population

Introduction and aimStudies have shown that two polymorphisms were associated with steatosis and progression of non-alcoholic fatty liver disease (NAFLD) in different populations: the Patatin-like Phospholipase Domain Containing 3 (PNPLA3) and Transmembrane 6 Superfamily Member 2 (TM6SF2). However, the frequency and significance of these polymorphisms in an admixed population, i.e., Brazilian, is unknown. Therefore, we aimed to evaluate them in healthy subjects in comparison to patients with NAFLD. Material and methodsThis was a multicenter cross-sectional study in 248 patients with biopsy-proven NAFLD and in 134 healthy controls from two tertiary centers in Brazil. PNPLA3 (rs738409 c.444C>G) and TM6SF2 (rs58542926 c.449C>T) polymorphisms were evaluated. ResultsIn controls, the frequencies of PNPLA3 CC and CG+GG were 49.25% and 50.74%, respectively; in NAFLD patients, this was 31.05% and 68.88% (p=0.0044, 95% CI 1.037–2.977). PNPLA3 GG subjects had an increased risk (3.29-fold) of having NAFLD when compared to CC subjects (p=0.0044, 95% CI 1.504–7.225). In patients with nonalcoholic steatohepatitis (NASH), PNPLA3 GG compared to CC was associated with higher AST levels [38.4±25.3 versus 36.7±40.1IU/L, p=0.0395)] and with the presence of liver fibrosis (≥F2 fibrosis, p=0.0272). TM6SF2 polymorphisms were not in Hardy-Weinberg equilibrium in our NAFLD group precluding further analysis. ConclusionWe demonstrated for the first time that PNPLA3 CG+GG increase the risk of NAFLD among Brazilian subjects. Moreover, PNPLA3 GG was associated with liver enzyme elevation and fibrosis in NASH patients.

Novel Interplay Between Sonic Hedgehog and Transforming Growth Factor-β1 in Human Nonalcoholic Steatohepatitis.

Nonalcoholic steatohepatitis (NASH) has the potential to progress to fibrosis, cirrhosis, and hepatocellular carcinoma. Upregulation of sonic hedgehog (Shh) has been documented in development of NASH through sustained cell stress. At the same time, transforming growth factor-β1 (TGF-β1), which is a central element in fibrogenic reactions in various diseases and sites, has been reported to be associated with hepatic inflammation and fibrotic reaction. To explore crosstalk between Shh and TGF-β1 in the development and progression of NASH, we investigated the expression of both these proteins in 135 human specimens of NASH, 35 fatty liver specimens, 35 specimens of alcoholic steatohepatitis with immunohistochemistry. Shh protein was expressed in the cytoplasm of ballooned hepatocytes with an ubiquitin-like pattern. In addition, a few scattered apoptotic hepatocytes in the inflammatory foci showed homogeneous cytoplasmic Shh expression. TGF-β1 protein was observed mainly in the activated hepatic stellate cells (HSCs) which were located in the inflammatory foci surrounding ballooned hepatocytes. Moreover, the mRNA levels of both Shh and TGF-β1 in the liver biopsy specimens from NASH patients was significantly increased compared with those in fatty liver patients. Statistically, there was a significant association of the expressions of Shh and TGF-β1 proteins in NASH (r=0.6, P<0.05). In addition, increased expression of Shh protein significantly parallels the severity of hepatocellular ballooning, lobular, and portal inflammatory responses and progression of fibrosis in NASH patients. Moreover, we found that much HSCs transformed into myofibroblast-like phenotype and migrated downward to HepG2 hepatocellular carcinoma cells with overexpression of Shh by transwell assay. We also observed overexpression of proteins of Shh and TGF-β1 in cultured activated HSCs with confocal microscopy. These findings strongly suggest there is interplay between Shh and TGF-β1 in hepatic inflammatory reactions. Shh secreted through damaged hepatocytes may result in activation of TGF-β1 and subsequent transformation of HSCs, which together modulate the progression of human NASH.

Cardiopulmonary protective effects of the selective FXR agonist obeticholic acid in the rat model of monocrotaline-induced pulmonary hypertension

Farnesoid X receptor (FXR) activation by obeticholic acid (OCA) has been demonstrated to inhibit inflammation and fibrosis development and even induce fibrosis regression in liver, kidney and intestine in multiple disease models. OCA also inhibits liver fibrosis in nonalcoholic steatohepatitis patients. FXR activation has also been demonstrated to suppress the inflammatory response and to promote lung repair after lung injury.This study investigated the effects of OCA treatment (3, 10 or 30mg/kg, daily for 5days a week, for 7 and/or 28 days) on inflammation, tissue remodeling and fibrosis in the monocrotaline (MCT)-induced pulmonary arterial hypertension (PAH) rat model. Treatment with OCA attenuated MCT-induced increased pulmonary arterial wall thickness and right ventricular hypertrophy, by i) blunting pathogenic inflammatory mechanisms (downregulation of interleukin 6, IL-6, and monocyte chemoattractant protein-1, MCP-1) and ii) enhancing protective mechanisms counteracting fibrosis and endothelial/mesenchymal transition. MCT-injected rats also showed a marked decrease of pulmonary artery responsiveness to both endothelium-dependent and independent relaxant stimuli, such as acetylcholine and a nitric oxide donor, sodium nitroprusside. Administration of OCA (30mg/kg) normalized this decreased responsiveness. Accordingly, OCA treatment induced profound beneficial effects on lung histology. In particular, both OCA doses markedly reduced the MCT-induced medial wall thickness increase in small pulmonary arteries. To evaluate the objective functional improvement by OCA treatment of MCT-induced PAH, we performed a treadmill test and measured duration of exercise. MCT significantly reduced, and OCA normalized treadmill endurance. Results with OCA were similar, or even superior, to those obtained with tadalafil, a well-established treatment of PAH.In conclusion, OCA treatment demonstrates cardiopulmonary protective effects, modulating lung vascular remodeling, reducing right ventricular hypertrophy and significantly improving exercise capacity. Thus, OCA can restore the balance between relaxant and contractile pathways in the lung, promoting cardiopulmonary protective actions.

Review article: emerging anti-fibrotic therapies in the treatment of non-alcoholic steatohepatitis.

Non-alcoholic fatty liver disease (NAFLD) can lead to non-alcoholic steatohepatitis (NASH) and cirrhosis. Fibrosis predicts worse outcomes and mortality. New treatments targeting fibrosis are being investigated to reverse disease progression. To review the new pipeline therapeutic agents targeting fibrosis in NASH patients, with particular focus on clinical trials in which reversing fibrosis and portal hypertension are the primary outcomes. The literature was searched in PubMed between January 2000 and January 2016 using search terms non-alcoholic fatty liver disease and NASH, with filters of 'English language'. We focused on fibrosis improvement as the key outcome. We also searched the ClinicalTrials.gov for promising agents that target fibrosis in NASH patients. Significant advances have been made on approaches targeting fibrosis in NASH patients. Many therapeutic agents are already in development, some of which have shown promising results in preclinical and phase I studies. Novel therapies have entered phase II and III studies targeting fibrosis reversal and/or improvement in portal hypertension. Innovative studies have also started looking into combining these agents, aiming at different mechanisms to maximise therapeutic outcomes. We found five clinical trials in phase II and one in phase III focusing on fibrosis in NASH patients as key outcomes. One of the phase II trials is using combination therapy to target fibrosis. Ongoing research studies are already investigating new pathways aimed at reversing fibrosis in NASH patients. Novel therapeutic agents are in development and are expected to offer unique options to NASH patients with advanced fibrosis.

Usefulness of Cytokeratin-18M65 in Diagnosing Non-Alcoholic Steatohepatitis in Japanese Population

Objective: The aim of this study was to evaluate cytokeratin-18M65 (CK-18M65) for distinguishing between simple steatosis (SS) and non-alcoholic steatohepatitis (NASH) against healthy individuals (HIs) in Japanese population. Methods: The serum from 24 HIs, 21 patients with SS and 20 patients with NASH were examined. Serum CK-18M65 was measured by enzyme-linked immunosorbent assay. Results: Aspartate aminotransferase was significantly different between NASH patients and HIs with p < 0.0001 (SS patients and HIs: p < 0.0001), as was alanine aminotransferase between NASH patients and HIs with p < 0.0001 (SS patients and HIs: p < 0.0001). Serum CK-18M65 increased in a stepwise fashion in HIs and also in SS and NASH patients. Multivariate logistic regression analysis revealed that NASH could be diagnosed with the use of CK-18M65 alone (p = 0.0285, OR 1.0038, 95% CI 1.0004-1.0073). At the optimal cut-off level of 548 U/l, CK-18M65 had an AUC value of 0.7369, 60.00% sensitivity and 85.70% specificity. In patients with NASH, no significant difference was observed between low fibrosis (Stage 0-1, 794.30 ± 454.41, n = 10) and high fibrosis (Stage 2-3, 809.70 ± 641.43, n = 10; p = 0.5967) and between slight steatosis (<33%, 512.89 ± 229.65, n = 9) and moderate steatosis (≥33%, 655.13 ± 480.78, n = 32) in patients with non-alcoholic fatty liver disease (NAFLD; p = 0.7647) with the use of CK-18M65. Conclusion: Serum CK-18M65 distinguished NASH from SS, but could not assess the severity of steatosis in NAFLD patients or the grade of fibrosis in NASH patients in Japanese population.

Total body weight loss of ≥ 10 % is associated with improved hepatic fibrosis in patients with nonalcoholic steatohepatitis.

Given the rising epidemics of obesity and metabolic syndrome, nonalcoholic steatohepatitis (NASH) is now the most common cause of liver disease in the developed world. Effective treatment for NASH, either to reverse or prevent the progression of hepatic fibrosis, is currently lacking. To define the predictors associated with improved hepatic fibrosis in NASH patients undergoing serial liver biopsies at prolonged biopsy interval. This is a cohort study of 45 NASH patients undergoing serial liver biopsies for clinical monitoring in a tertiary care setting. Biopsies were scored using the NASH Clinical Research Network guidelines. Fibrosis regression was defined as improvement in fibrosis score ≥1 stage. Univariate analysis utilized Fisher's exact or Student's t test. Multivariate regression models determined independent predictors for regression of fibrosis. Forty-five NASH patients with biopsies collected at a mean interval of 4.6 years (±1.4) were included. The mean initial fibrosis stage was 1.96, two patients had cirrhosis and 12 patients (26.7 %) underwent bariatric surgery. There was a significantly higher rate of fibrosis regression among patients who lost ≥10 % total body weight (TBW) (63.2 vs. 9.1 %; p = 0.001) and who underwent bariatric surgery (47.4 vs. 4.5 %; p = 0.003). Factors such as age, gender, glucose intolerance, elevated ferritin, and A1AT heterozygosity did not influence fibrosis regression. On multivariate analysis, only weight loss of ≥10 % TBW predicted fibrosis regression [OR 8.14 (CI 1.08-61.17)]. Results indicate that regression of fibrosis in NASH is possible, even in advanced stages. Weight loss of ≥10 % TBW predicts fibrosis regression.

Liver ultrasound scanning in the detection of hepatic steatosis and fibrosis in NASH patients

Abstract Background Nonalcoholic fatty liver disease (NAFLD) has become the most prevalent cause of liver disease in western countries. The development of nonalcoholic steatohepatitis (NASH) and fibrosis identifies the risk group with an increased incidence of liver-related deaths. Aim The aim of the present study is to investigate how accurately liver ultrasound (US) can contribute toward the prediction of the severity of liver damage in NAFLD, and to determine whether it may be an easily available, inexpensive, noninterventional, widely used screening method. Methods Fifty-four obese patients with variable degrees of BMI were recruited in the present study. Assessment of full medical history, anthropometric measurements, biochemical studies, abdominal US, liver biopsy for histological examination, and determination of the NAFLD activity score (NAS) score were carried out on all patients to identify NASH patients . Liver steatosis was evaluated using liver US, and graded according to a semiquantitative scale from 1 to 4. Liver histological examination was carried out to identify patients with NASH, borderline NASH, or non-NASH according to the NAS score. Results According to the NAS score, patients were divided into non-NASH patients (eight patients), borderline NASH patients (24 patients), NASH patients (20 patients), and patients with NASH and fibrosis (two patients). Alanine aminotransferase and γ-glutamyl transpeptidase were significantly higher in NASH patients. Correlating the grading of hepatic steatosis by liver US and NAS score, grade 1 was found in 37.5% of patients with non-NASH, 33.3% of patients with borderline NASH, and only in 5% of patients with NASH, whereas grade 4 steatosis was found in 20% of NASH patients and 4.2% of patients with borderline NASH; none of the non-NASH patients were diagnosed with grade 4 hepatic steatosis. The sensitivity of liver US in detecting grades of steatosis in liver biopsy was 61% in grade 1, 25% in grade 2, and 75% in grade 3. There was a direct correlation between grading of steatosis in the histological examination and the presence of NASH, P less than 0.000. Conclusion Liver US is not only sensitive in the detection of hepatic steatosis, but also in the prediction of the presence of NASH; therefore, it can be used as a simple, noninvasive, low-cost method for the screening of NAFLD and for the early identification of patients in need of aggressive intervention.

Liver Elasticity in NASH Patients Evaluated with Real-Time Elastography (RTE)

Liver elasticity as assessed by real-time elastography (RTE) has been shown to be correlated to liver fibrosis in various chronic liver diseases. The aim of our study was to assess the RTE performance in the evaluation of liver fibrosis in nonalcoholic steatohepatitis (NASH), as well as the histopathologic variables determining the eventual discordance between the RTE-predicted and the biopsy-proven fibrosis. Fifty-two consecutive biopsy proven NASH patients and 20 controls were studied. Liver tissue elasticity measurements were performed using the Hitachi EUB-8500 sonographer and the EUP-L52 Linear (3–7 MHz) probe. RTE liver tissue mean elasticity (TME) values were calculated and correlated to the histologic fibrosis, activity and steatosis scores. A decrease in TME was observed with increasing fibrosis (r = −0.75). Similarly, TME varied together consistently with steatosis (r = −0.3). In contrast, TME did not show any correlation with the severity of inflammation. Multiple regression analysis showed that fibrosis was the only variable able to significantly (p < 0.0001) modify TME values. The diagnostic accuracy of TME measurement for F > 0 evaluated by AUC-ROC analysis was 0.86. The diagnostic accuracy of TME measurement for F ≥ 2 was 0.92. We suggest that RTE could be used as a complementary imaging method to evaluate liver fibrosis in NASH patients. Future studies of larger patient cohorts are necessary for the validation of the technique.

Serum TGF-β, Serum MMP-1, and HOMA-IR as non-invasive predictors of fibrosis in Egyptian patients with NAFLD

Background/Aim:Non-alcoholic fatty liver disease (NAFLD) is an increasingly prevalent cause of chronic liver disease worldwide. A number of these patients progress to nonalcoholic steatohepatitis (NASH) which carries significant morbidity and mortality. The aim of this study is to evaluate the diagnostic value of serum levels of transforming growth factor beta-1 (TGF-β1) matrix metalloproteinase-1 (MMP-1), and insulin resistance as predictors of fibrosis in Egyptian NAFLD patients.Patients and Methods:Fifty patients with NAFLD and different stages of fibrosis were studied. Serum levels of TGF-β1, MMP-1, and fasting serum insulin were measured; calculation of the homeostasis model assessment for insulin resistance (HOMA-IR) was done.Results:TGF-β1 gives a sensitivity of 100% and specificity of 94.4% for stage 1 fibrosis, 100% and 93.9%, respectively, for stage 2 fibrosis, and 97.7% and 100%, respectively, for stage 3 fibrosis. MMP-1 showed sensitivity and specificity of 88% and 81.8%, respectively, for stage 2 fibrosis, 90.9% and 55.56%, respectively, for stage 3 fibrosis, but it is of no diagnostic value in stage 1 fibrosis.Conclusion:Serum TGF-β1, MMP-1, and insulin resistance (HOMA-IR) proved to be potentially useful noninvasive markers in predicting fibrosis in NASH patients.

Differential expression of miRNAs in the visceral adipose tissue of patients with non‐alcoholic fatty liver disease

Progression of non-alcoholic fatty liver disease (NAFLD) can be facilitated by soluble molecules secreted by visceral adipose tissue (VAT). MicroRNAs (miRNAs) are likely to regulate some of these molecular pathways involved in pathogenesis of NAFLD. To profile miRNA expression in the visceral adipose tissue of patients with NAFLD. Visceral adipose tissue samples were collected from NAFLD patients and frozen. Patients with biopsy-proven NAFLD were divided into non-alcoholic steatohepatitis (NASH) (n = 12) and non-NASH (n = 12) cohorts controlled for clinical and demographic characteristics. Extracted total RNA was profiled using TaqMan Human MicroRNA arrays. Univariate Mann-Whitney comparisons and multivariate regression analysis were performed to compare miRNA profiles. A total of 113 miRNA differentially expressed between NASH patients and non-NASH patients (P < 0.05). Of these, seven remained significant after multiple test correction (hsa-miR-132, hsa-miR-150, hsa-miR-433, hsa-miR-28-3p, hsa-miR-511, hsa-miR-517a, hsa-miR-671). Predicted target genes for these miRNAs include insulin receptor pathway components (IGF1, IGFR13), cytokines (CCL3, IL6), ghrelin/obestatin gene, and inflammation-related genes (NFKB1, RELB, FAS). In addition, two miRNA species, hsa-miR-197 and hsa-miR-99, were significantly associated with pericellular fibrosis in NASH patients (P < 0.05). Levels of IL-6 in the serum negatively correlated with the expression levels of all seven miRNAs capable of down regulating IL-6 encoding gene. miRNA expression from VAT may contribute to the pathogenesis of NAFLD - a finding which may distinguish relatively simple steatosis from NASH. This could help identify potential targets for pharmacological treatment regimens and candidate biomarkers for NASH.