Wnt-induced secreted protein 1/CCN4 in liver fibrosis both in vitro and in vivo.

BACKGROUND: Wnt-induced secreted protein-1 (WISP-1/CCN4) is a member of the CCN family growth factors, and its role in liver fibrosis is largely unknown. METHODS: For in vitro, hepatic stellate cells (HSCs) were isolated from Sprague-Dawley rats. Expression of WISP-1 during progressive activation of cultured rat HSCs was analyzed by qRT-PCR. The effects of TNF-a and TGF-beta1 on WISP-1 expression were analyzed in stellate cell lines HSC-T6 and LX-2. The effect of exogenous WISP-1 protein on LX-2 proliferation was examined. For in vivo, expressions of WISP-1 in fibrotic liver of a carbon tetrachloride (CCl4)-induced fibrosis rat model were analyzed by qRT-PCR and immunohistochemistry. RESULTS: In vitro, WISP-1 was increasingly expressed during progressive activation of cultured rat HSCs. WISP-1 was significantly induced in HSC-T6 cells by TNF-a and in LX-2 cells by TGF-beta1. Recombinant WISP-1 protein promoted LX-2 proliferation in a dose-dependent manner. In vivo, both mRNA and protein expression levels of WISP-1 were increased significantly in experimental hepatic fibrosis model. CONCLUSIONS: Our results showed the upregulation of WISP-1 in both in vitro and in vivo liver fibrosis models, and WISP-1 stimulated the proliferation of HSCs in vitro. These results may be helpful to elucidate the exact role of WISP-1 in liver fibrogenesis.

Noninvasive prediction of large esophageal varices in liver cirrhosis patients.

PURPOSE: Esophageal varices are a dangerous complication of liver cirrhosis. The development of cost effective, noninvasive means for prediction of large esophageal varices could reduce the use of upper gastrointestinal endoscopy in variceal screening and also provide an alternative way to confirm the results of conventional endoscopic diagnosis. Previously proposed predictive models are neither sensitive nor specific. METHODS: A retrospective study based on a group of 104 liver cirrhosis patients was performed. Multiple statistical approaches were used to evaluate the association of large esophageal varices with 20 individual and six compound clinical laboratory variables. A new predictive model was developed. RESULTS: Univariate analysis suggested that eight out of 26 variables were significantly associated with large esophageal varices. Further stepwise logistic regression eventually identified three variables (hemoglobin level, portal vein diameter and the ratio of platelet count/spleen diameter) that contributed significantly to the final regression model. Receiver operating characteristic (ROC) curve analysis showed that this new regression model achieved 77.8% and 72% of diagnostic sensitivity and specificity, respectively, for the prediction of large esophageal varices. In our study group, its diagnostic accuracy (AUROC=0.814) was found to be significantly higher than six predictive models previously published. CONCLUSIONS: No single variable offers self-sufficient predictive function for large esophageal varices. A comprehensive model using multiple variables significantly improves the predictive accuracy in screening the most at risk patients with potential variceal hemorrhage.

The effect of carvedilol and propranolol on portal hypertension in patients with cirrhosis: a meta-analysis.

PURPOSE: Several randomized controlled clinical trials have been conducted to investigate the role of carvedilol and propranolol on the effect of portal pressure in patients with cirrhosis, leading to controversial results. Current meta-analysis was performed to compare the efficacy of the two drugs on portal pressure. PATIENTS AND METHODS: Two-hundred and ninety eligible patients were recruited. Published studies were selected based on PubMed, the Cochrane Library, Chinese Journal Full-text Database, and Wanfang Database. The outcome measurements included the mean difference (MD) in the percentage of hepatic vein pressure gradient reduction (%HVPG reduction), the risk ratio (RR) of nonresponders in hemodynamic assessment, and the percentage of mean arterial pressure reduction (%MAP reduction). Subgroup analysis was performed. RESULTS: Seven trials were identified (including five acute and three long-term drug administration randomized controlled trials). A summary of pooled MD between the %HVPG reduction is as follows: overall -8.62 (confidence interval [CI] -11.76, -5.48, P<0.00001), acute -10.05 (CI -14.24, -5.86, P<0.00001), and long term -6.80 (CI -11.53, -2.07, P=0.005), while summary of pooled RR of hemodynamic nonresponders with carvedilol was as follows: overall 0.64 (CI 0.51, 0.81, P=0.0002), acute 0.63 (CI 0.47, 0.85, P=0.002), and long term 0.67 (CI 0.47, 0.97, P=0.03). Both of the outcome measurements favored carvedilol. Significant heterogeneity (P<0.1, I (2)=92%) existed between the two treatment groups in %MAP reduction. No considerable difference could be observed in the %MAP reduction through the poor overlapping CI boundaries. CONCLUSION: Carvedilol has a greater portal hypertensive effect than propranolol. Further comparative trials of the two drugs are required to identify the effect of MAP reduction.

Focal peliosis hepatis in a colon cancer patient resembling metastatic liver tumor.

Peliosis hepatis (PH) is a rare benign condition characterized by the presence of multiple, randomly distributed, blood filled cystic areas of variable size within the liver parenchyma. PH is difficult to recognize and may be mistaken for neoplasm, metastases or multiple abscesses. A 75-year-old female with a previous history of colon cancer was admitted when a liver mass in the right liver lobe was found 11 mo after surgery during the follow-up period. Computed tomography and magnetic resonance imaging scan of the abdomen were performed. The initial possible diagnosis was metastatic hepatocellular carcinoma. The patient underwent excision of the hepatic segment where the nodule was located. The pathological diagnosis of the surgical specimen was PH. PH should be considered in the differential diagnosis of new liver lesions in patients whose clinical settings do not clearly favor metastasization. Clinicians and radiologists must recognize these lesions to minimize the probability of misdiagnosis and inappropriate treatment.

Effect of oxymatrine on hepatic gene expression profile in experimental liver fibrosis of rats.

OBJECTIVE: To investigate the effects of oxymatrine on hepatic gene expression profile in a rat model of liver fibrosis. METHODS: Forty healthy male SD rats were randomly divided into three groups, a normal group (n=8), a model group (n=16), and an oxymatrine treatment group (n=16). Experimental hepatic fibrosis was induced by subcutaneous injection of carbon tetrachloride (CCl(4)). The rats in the treatment group received oxymatrine via celiac injection at a dosage of 40 mg/kg once a day at the same time. The rats in the model and normal groups received saline at the same dosage via celiac injection. Serum levels of aspartate aminotransferase (AST), alanine transaminase (ALT), alkaline phosphatase (AKP), hyaluronic acid (HA), and laminin (LN) were assayed. The deposition of collagen was observed with HE and Masson staining. Effect of oxymatrine on hepatic gene expression profile was detected by oligonucleotide microarray analysis with Affymetrix gene chip rat U230A. Quantitative real-time polymerase chain reaction (QRT-PCR) was carried out to confirm the expression changes of six genes. RESULTS: Oxymatrine significantly improved liver function, lowered serum levels of HA and LN, and decreased the degree of liver fibrosis, compared with the model group (P<0.05). A total of 754 differentially expressed genes were identified by gene chip between the model group and the normal group, among which 438 genes increased and 316 genes decreased over two folds. Compared with the model group, 86 genes were downregulated markedly in the oxymatrine group (P<0.05), including collagen I and other genes related to extracellular material (ECM), integrin signal transduction genes, early growth response factor genes, and proinflammatory genes; 28 genes were upregulated significantly (P<0.05), including cytochrome P450 (CYP450) superfamily genes, glycolipids metabolism and biological transformation related genes. Six genes were confirmed with QRT-PCR, consistent with the result from microarray. CONCLUSION: Oxymatrine could affect the expression of many functional genes and may be useful in the prevention and treatment of liver fibrosis.

Wnt5a participates in hepatic stellate cell activation observed by gene expression profile and functional assays.

AIM: To identify differentially expressed genes in quiescent and activated hepatic stellate cells (HSCs) and explore their functions. METHODS: HSCs were isolated from the normal Sprague Dawley rats by in suit perfusion of collagenase and pronase and density Nycodenz gradient centrifugation. Total RNA and mRNA of quiescent HSCs, and culture-activated HSCs were extracted, quantified and reversely transcripted into cDNA. The global gene expression profile was analyzed by microarray with Affymetrix rat genechip. Differentially expressed genes were annotated with Gene Ontology (GO) and analyzed with Kyoto encyclopedia of genes and genomes (KEGG) pathway using the Database for Annotation, Visualization and Integrated Discovery. Microarray data were validated by quantitative real-time polymerase chain reaction (qRT-PCR). The function of Wnt5a on human HSCs line LX-2 was assessed with lentivirus-mediated Wnt5a RNAi. The expression of Wnt5a in fibrotic liver of a carbon tetrachloride (CCl(4))-induced fibrosis rat model was also analyzed with Western blotting. RESULTS: Of the 28 700 genes represented on this chip, 2566 genes displayed at least a 2-fold increase or decrease in expression at a P < 0.01 level with a false discovery rate. Of these, 1396 genes were upregulated, while 1170 genes were downregulated in culture-activated HSCs. These differentially expressed transcripts were grouped into 545 GO based on biological process GO terms. The most enriched GO terms included response to wounding, wound healing, regulation of cell growth, vasculature development and actin cytoskeleton organization. KEGG pathway analysis revealed that Wnt5a signaling pathway participated in the activation of HSCs. Wnt5a was significantly increased in culture-activated HSCs as compared with quiescent HSCs. qRT-PCR validated the microarray data. Lentivirus-mediated suppression of Wnt5a expression in activated LX-2 resulted in significantly impaired proliferation, downregulated expressions of type I collagen and transforming growth factor-ß1. Wnt5a was upregulated in the fibrotic liver of a CCl(4)-induced fibrosis rat model. CONCLUSION: Wnt5a is involved in the activation of HSCs, and it may serve as a novel therapeutic target in the treatment of liver fibrosis.