The recruitment of TNF receptor–associated factor 2 (TRAF2) media

The recruitment of TNF receptor–associated factor 2 (TRAF2) mediates the proinflammatory consequences of CD154/CD40 interaction.61, 62 As IRE1 recruits TRAF2 upon activation, TRAF2 may represent a potential link between the CD40 and IRE1 signalization pathways. Our study does not exclude other mechanisms through which CD154 may

interfere with the progression of liver steatosis. These may involve deregulation of the cytokine network. Indeed, CD154 induces inflammatory cytokines, some of which play a role in lipid metabolism, such as IL-6. IL-6 alleviates liver steatosis63 and IL-6−/− mice develop mature-onset obesity and are prone to hepatic steatosis and metabolic alterations.64, 65 According to the regulatory role of CD154 on IL-6 expression, we found that CD154KO PI3K Inhibitor Library purchase mice showed impaired induction of IL-6 following the olive oil–rich diet as shown by a reduced induction of plasma IL-6 levels and liver IL-6 mRNA (Supporting Fig. 9A,B). Hence, the down-regulation of IL-6 expression may provide another mechanism to explain the steatotic phenotype of olive oil–fed CD154KO mice. ER stress also leads to IL-6 production through XBP-1 signaling59, 66 and, accordingly, in HepG2 cells expressing a dominant negative form of IRE1, TM-induced expression of IL-6 was impaired. In this context, the CD154-dependent IL-6 induction selleck monoclonal humanized antibody inhibitor was preserved (Supporting Fig. 9A,B). Therefore, the control

of IL-6 expression is likely to represent another interface linking CD154, the UPR, and hepatic lipid metabolism. This observation suggests that several integrated signaling pathways are likely to account for the contribution of CD154 Urease in hepatic steatosis. In conclusion, our study shows that CD154 is a mediator involved in the natural history of hepatic steatosis. CD154 appears as a new link between lipid metabolism and inflammation in the liver, supporting the idea of interdependency between inflammation and metabolic disorders.27, 32 The authors thank Chantal Combe, Jérôme Gabet, Alexandra Nicou, and Antonio Palos Pinto for technical help. Additional Supporting Information may be found in the online version of this

article. “
“See article in J. Gastroenterol. Hepatol. 2012; 27: 789–796. Nonalcoholic fatty liver disease (NAFLD) is now the most common cause of chronic liver disease worldwide. Nonalcoholic steatohepatitis (NASH) is a progressive form of NAFLD with the potential for progression to cirrhosis. The pathogenesis of NASH is incompletely understood, but may involve hyperendotoxemia1 secondary to impaired phagocytotic function of Kupffer cells (KCs)2 and consequent KC overproduction of and increased sensitivity to cytokines such as tumor necrosis factor (TNF)-α and interleukin-1β (IL-1β).3 Impaired phagocytotic function of KCs may therefore lead to higher endotoxin levels in the systemic circulation, as has been observed in patients with NASH and in animal models of NASH.

Twelve studies were retrospective, observational and non-interven

Twelve studies were retrospective, observational and non-interventional studies. According to our meta-analysis, the rate of serological HBsAg− and anti-HBc+ was higher among HCC patients compared with non-HCC patients (odds ratio [OR], 1.55; 95% CI, 1.22–1.98). HCV patients that were anti-HBc+ had a greater chance of developing HCC than their anti-HBc− counterparts (OR, 2.15; 95% CI, 1.34–3.47). Conclusions:  The serological status of HBsAg− and anti-HBc+ appears to be correlated with a poor prognosis for chronic

HCV infection. Though the general quality of these references was low, and multiple confounding factors existed, the likelihood of a poorer outcome of HCV patients that are positive for anti-HBc should be considered by their physicians. “
“Hepatitis C virus (HCV) directly induces RO4929097 cell line oxidative stress and liver injury. Bach1, a basic leucine zipper mammalian transcriptional repressor, negatively regulates AZD2014 supplier heme oxygenase 1 (HMOX1), a key cytoprotective enzyme that has antioxidant and anti-inflammatory activities. microRNAs (miRNAs) are small noncoding RNAs (≈22 nt) that are important regulators of gene expression. Whether

and how miRNAs regulate Bach1 or HCV are largely unknown. The aims of this study were to determine whether miR-196 regulates Bach1, HMOX1, and/or HCV gene expression. HCV replicon cell lines (Con1 and 9–13) of the Con1 isolate and J6/JFH1-based HCV cell culture system were used in this study. The effects of miR-196 mimic on Bach1, HMOX1, and HCV RNA, and protein levels were measured by way of quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting, respectively. The Dual Glo Luciferase Assay System was used to determine reporter activities. miR-196 mimic significantly down-regulated Bach1 and up-regulated HMOX1 gene expression and inhibited HCV expression. Dual luciferase reporter assays demonstrated that transfection

of miR-196 mimic resulted in a significant decrease in Bach1 3′-untranslated region (UTR)–dependent luciferase activity but not in mutant Bach1 3′-UTR–dependent luciferase activity. Moreover, there was no detectable effect of mutant miR-196 on Bach1 3′-UTR–dependent luciferase activity. Conclusion: miR-196 directly acts on the 3′-UTR of Bach1 messenger RNA and translationally represses the expression Mannose-binding protein-associated serine protease of this protein, and up-regulates HMOX1. miR-196 also inhibits HCV expression in HCV replicon cell lines (genotype 1b) and in J6/JFH1 (genotype 2a) HCV cell culture system. Thus, miR-196 plays a role in both HMOX1/Bach1 expression and the regulation of HCV expression in human hepatocytes. Overexpression of miR-196 holds promise as a potential novel strategy to prevent or ameliorate hepatitis C infection, and to protect against liver injury in chronic HCV infection. (HEPATOLOGY 2010.) Hepatitis C virus (HCV) infection is a worldwide health problem.

The data available to date have implications for the use of BAY 9

The data available to date have implications for the use of BAY 94–9027 in persons with haemophilia. They suggest that keeping an open mind and vigilance are key aspects of ongoing and future clinical trials as the risk-benefit profiles evolve for these compounds. Findings from this literature of the safety and elimination of high molecular weight PEGylated proteins (PEG ≥ 30 kDa) are consistent with the toxicology study data presented, herein, for BAY 94–9027 as well as the 60 kDa PEG moiety. The data summarized, herein, indicate that long-term treatment with BAY 94–9027 is not expected

to result in an increased safety risk due to PEG. The currently available information shows a lack of preclinical toxicity for high molecular weight PEG molecules currently used in the mono-PEGylation SB203580 cell line of therapeutic PS-341 proteins. Data indicates that long-term chronic treatment with a 60 kDa PEGylated rFVIII would appear to be safe and should not result in PEG-related adverse events, specifically,

as the anticipated clinical dose of BAY 94–9027 contains only a very small amount of PEG. Confirmatory results from clinical trials are needed to support these conclusions. There is also need for more research regarding the long-term safety of modified coagulation products and the comparative safety of different biologics. National and international registries and other types of large databases are relevant sources for providing complementary evidence regarding the short- and longer term safety of these products. The authors thank Andrea Loewe, Anita Shah, Elke Dittrich-Wengenroth, Julia Franco, Klaus Buehner, Bernhard Beckermann and Friedrich-Wilhelm Jekat for their support, for their discussions and valuable input to this publication. We also thank the members of the Bayer International advisory board, and the Bayer US Hemophilia Council for their insight into the discussion of PEGylated proteins. Inge A. Ivens, Suplatast tosilate Prasad Mathew – Performed the research, analysed the data, wrote the paper. Andreas Baumann,

Thomas McDonald, Thomas Humphries, and Lisa Michaels – analysed the data, critiqued the paper and contributed to the acquisition, analysis and interpretation of data. All authors have approved the version of this manuscript. The authors are employees of Bayer Health Care Pharmaceuticals and Bayer Pharma AG. “
“Summary.  Establishing haemostasis for surgical procedures in children with inherited bleeding disorders is challenging. Providers are often hesitant to undertake surgeries in children with bleeding disorders out of fear of bleeding complications. To review the preoperative management and haemorrhagic complications of children with inherited bleeding disorders at our institution, we conducted a retrospective electronic medical record review from 1999 to 2010.

To quantitate relative AP2 membrane staining, random fields were

To quantitate relative AP2 membrane staining, random fields were visualized by epifluorescence and digitized. From micrographs, membrane fluorescence was traced using the segmented line tool, and intracellular staining

regions of interest (ROI) were measured using the ImageJ Measure ROI tool. The averaged background pixel intensity was subtracted from both the averaged membrane and intracellular intensities, and the ratio of basolateral versus intracellular fluorescence intensity was determined. The K+ depletion/repletion assays were performed as previously described.22 For ASGP-R antibody trafficking studies, K+-depleted cells Smoothened Agonist cell line were surface labeled with anti-ASGP-R antibodies (1:25) for 20 minutes on ice. Cells were reincubated in prewarmed medium supplemented with 10 mM of KCl, and the ASGP-R antibody-antigen complexes

were allowed to traffic for desired times at 37°C. Cells were fixed, permeabilized, and the trafficked ASGP-R antibodies were labeled with secondary antibodies. Cells were stained as described above and mounted in Tris-buffered saline (pH 10.5) containing 5% glycerol and 4 mg/mL of phenylenediamine. Fluorophores were excited with a 2.5-W Kr/Ar laser Lapatinib mouse (Spectra Physics, Irvine, CA) and visualized using an Olympus 1X 71 inverted microscope and total internal reflection fluorescence (TIRF) illuminator (Olympus, Center Valley, PA). Images were collected using a Photometrics Evolve EM-CCD (charge-coupled

device) camera (Photometrics, Tuscon, AZ) and Metamorph software (Molecular Devices, Sunnyvale, CA). Puncta were counted using the FociPicker three-dimensional ImageJ plugin. Fully covered 10,000 px2 ROIs were selected from random images. In general, five images/experiment were acquired and two to five fields/image were counted. For transmission electron microscopy (TEM), cells were fixed and processed using standard Epon embedding techniques. Ultrathin sections were cut and stained with uranyl acetate, followed by lead citrate. Grids were viewed on a Hitachi 7600 transmission electron microscope (Hitachi, Tokyo, Japan), and images were captured with an AMT CCD camera (Advanced Microscopy Techniques, Woburn, from MA). We previously determined that ethanol exposure led to the dramatic redistribution of ASGP-R from intracellular endosomes to the basolateral membrane in WIF-B cells.15 Closer examination using confocal microscopy revealed that the membrane-associated ASGP-R in ethanol-treated cells was present in discrete puncta (Fig. 1A). Because these puncta resembled clathrin-coated pits visualized at the light level, we examined the distributions of core clathrin-coat proteins. In control cells, CHC localized primarily to an intracellular compartment (Fig. 1A). As observed for ASGP-R, CHC redistributed to the basolateral membrane in discrete puncta in ethanol-treated cells.

Methods — In this observational study, adult patients with migrai

Methods.— In this observational study, adult patients with migraine

were consecutively recruited. Disability was measured with the MIDAS (Migraine Disability Assessment) and the WHO-DAS II (World Health Organization Disability Assessment Schedule), HRQoL with the SF-36 (Medical Outcome Survey 36-item Short-Form Health Survey). Spearman’s rank correlation between MIDAS score, CH5424802 datasheet SF-36 and WHO-DAS II scales was performed to evaluate the relationships between quality of life and disability. The impact of migraine on disability and HRQoL was assessed by comparing WHO-DAS II and SF-36 scores against Italian normative values, and by evaluating the different disability and HRQoL profiles in patients with different find more severity of migraine, defined according to migraine frequency and pain intensity. Results.— A total of 102 patients with migraine (87 females)

were enrolled. Mild to moderate correlations were reported between WHO-DAS II and SF-36′s PCS (r = −0.67, P < .01) and MCS (r = −0.36, P < .05) scales; MIDAS score correlations to SF-36's PCS (r = −0.44, P < .01) and MCS (not significant) were lower than WHO-DAS II summary score. The correlation between MIDAS score and the WHO-DAS II summary score was mild (r = −0.36, P < .05). The majority of HRQoL and disability scales (with the exception of SF-36's Physical Functioning, and WHO-DAS II Getting along with people scales) scored significantly lower than normative values. A trend towards worsening of both HRQoL and NADPH-cytochrome-c2 reductase disability, consistent with increasing migraine severity, was reported (Mann-Whitney’s U = 119.5 for MIDAS; U = 113.0 for WHO-DAS II summary score, both with P < .01; U = 152.9 for PCS; U = 171.0 for MCS, both with P < .05) Conclusions.— In migraineurs attending an Italian specialty headache clinic, disability scores were worse and HRQoL scores lower than those of the general population, and worsened consistently with increased

migraine severity. Measures of HRQoL and disability evaluate different psychosocial aspects of migraine and researchers should continue to employ them in public health and clinical research on migraine. They provide information on a poorly recognized part of migraine’s burden, where economic impact is minimal but there are important effects on patients’ daily lives in terms of interpersonal relationships, perceived quality of life and emotional status. “
“Nutrition must affect the structure and functioning of the brain. Since the brain has very high metabolic activity, what we consume throughout the day is likely to dramatically influence both its structure and moment to moment function.

Denervated transplanted livers lack acetylcholine modulation of p

Denervated transplanted livers lack acetylcholine modulation of proliferation of cells lining the canals of Hering. Hepatitis-injured transplanted livers also exhibit lower numbers of progenitor and reactive ductular cells than innervated matched controls.

Experiments in rats with galactosamine-damaged livers confirm that vagotomy induces impaired regeneration of progenitors and ductal reaction in cholangiocytes.43 Mechanotransduction mechanisms are another major set affecting lineage biology, most involving cytoskeletal rearrangements. The cytoskeleton is a ubiquitous cellular component with characteristics of amplification systems and connections with matrix. Some of these connections allow cells to sense microenvironment rigidity through nonmuscle myosin II, which

directs stiffness-dependent STI571 research buy differentiation in mesenchymal stem cells.44 Germ layer organization and cell sorting depends on cell adhesion forces and cortex tension relying on actomyosin network activity.45 Integrins connect the cytoskeleton to matrix substrata, recruit focal adhesions that adapt cells to mechanical stresses, bind ligands, and regulate intracellular signaling.46 Mechanical stretch in liver cells induces activation and synthesis of morphogens in the transforming growth factor beta (TGF-β) family of Activin/Nodal signaling.47 SMAD transcription factors regulate TGF-β signaling pathways and regulate gene expression through kinesin-mediated nucleocytoplasmic shuttling along intact microtubules.48, 49 Primary cilia in cells from soft organs also participate in mechanotransduction Pembrolizumab cost by probing and amplifying the effects of intraluminal flow above the cells apical surfaces. They mediate polarized signal transduction pathways that use the cytoskeleton to ensure specific and nondiffusable Reverse transcriptase signal trafficking to the nucleus.50 PDGRα and Hedgehog signaling take place in primary cilia51, 52 in livers of all ages5 through dynein-mediated shuttling of Gli transcription factors.53 Some chromatin targets of Gli transcription factors

include PTCH, WNT, and BMP genes, all involved in embryonic development and differentiation mechanisms.54-56 Hedgehog expression gradients also demarcate the extension of endodermal organs during development.52, 57 In conjunction, this information suggests primary cilia are relevant participants in endoderm maturation and differentiation. Bile secretion is an important mechanism for homeostatic control of tissue mass, operating as an inductor in mechanotransduction. Bile is a Newtonian fluid in normal physiological conditions with salt concentration-dependent viscosity.58 As hepatic parenchyma perform secretory functions; bile tonicity increases while flowing in the pericentral-to-periportal direction. Abnormal bile tonicity is characteristic of pathological conditions.

We found a significant interaction between HOMA-IR and ethnicity

We found a significant interaction between HOMA-IR and ethnicity (P < 0.001), and, because of this interaction, we examined the effect of HOMA-IR on NASH separately for Latinos and non-Latino whites, while adjusting for the variables selected from the stepwise logistic regression model (see

below). Interaction between HOMA-IR and ethnicity remained statistically significant when diabetic participants were excluded from the analyses (data not shown). Multivariate logistic regression: The results from the multivariate logistic regression analysis are show in Table 5. Factors positively associated with NASH included female gender (P = 0.001), AST (P < 0.0001), diabetes mellitus (P = 0.01), hypertension (P = 0.02), and triglyceride level (P = 0.02). Platelet count (P = 0.006) was negatively associated with selleck products NASH histology. We also found significant effect modification between ethnicity and HOMA-IR, with HOMA-IR being a significant risk factor for NASH among non-Latino whites (odds ratio [OR], 1.06; 95% CI: 1.01-1.1), but not among Latinos (OR, 0.93; 95% CI: 0.85-1.02) (Fig. 1). We investigated associations between advanced

fibrosis and clinical, laboratory, and sociodemographic factors among non-Latino whites and Latinos using univariate and multivariate logistic regression analyses. Univariate logistic regression: Univariate logistic regression demonstrated that the following risk factors were significantly associated with advanced fibrosis: ethnicity, age, gender, MLN0128 ic50 education level, income, hypertension, diabetes, metabolic syndrome, BMI, WC, SBP, DBP, AST, ALT, GGT, alkaline phosphatase, albumin, platelets,

LDL, HOMA-IR, and palmar erythema. We found no significant evidence for effect modification of ethnicity on patient characteristics with respect to advanced fibrosis. Multivariate logistic regression: Results from the multivariate logistic regression analysis are shown in Table 6. Factors positively associated with advanced fibrosis included age (P = 0.01), female gender Adenosine (P = 0.03), AST (P = 0.001), alkaline phosphatase (P = 0.002), hypertension (P = 0.0005), and HOMA-IR (P < 0.0001). Platelet count (P < 0.0001), ALT (P = 0.004), and total cholesterol (P = 0.004) were significantly inversely associated with risk for advanced fibrosis. The large NASH CRN cohort of patients with well-characterized, biopsy-proven disease allowed for a detailed analysis of the associations of ethnicity and race with clinical and histological features of NAFLD. We found that, among individuals with NASH histology, Latinos were younger, consumed more carbohydrate calories, and engaged in less physical activity, compared to non-Latino whites. Additionally, Latinos with NASH had lower income and lower prevalence of hypertension, compared to non-Latino whites, which may be a reflection of similar ethnic trends in the general U.S. adult population with respect to these two characteristics.

15 Therefore, we determined the TGF-β1 levels in the tumors of th

15 Therefore, we determined the TGF-β1 levels in the tumors of the different genotypes. A TGF-β1 ELISA was performed on lysates prepared from tumors and normal liver tissue (Fig. 4A,B). Low levels of TGF-β1 were detected

in the normal Control and Tgfbr2KO livers. Assessment of TGF-β1 levels in normal Trp53KO liver tissue demonstrated a small, but significant, increase over normal liver from the Tgfbr2KO mice (P = 0.0357). TGF-β1 levels were further increased in Trp53KO tumor tissue compared with normal Trp53KO liver (P = 0.0079). Comparison of TGF-β1 levels RO4929097 order in Trp53KO tumors versus Trp53KO;Tgfbr2KO tumors revealed that Trp53KO tumors have higher levels of TGF-β1 than Trp53KO;Tgfbr2KO tumors (P = 0.0079). These findings suggest that TGF-β signaling in the setting of p53 deletion may help promote tumor formation by inducing TGF-β1 expression. Because TGF-β1 levels were increased in Trp53KO tumors, we assessed the activation status of TGF-β signaling pathways in these tumors, including both Smad-dependent and Smad-independent pathways (Fig. 5). Immunoblot and immunohistochemistry analysis of liver tissue find more from both Trp53KO and Trp53KO;Tgfbr2KO mice detected the expression of phospho-Smad2 in both tumor genotypes (Supporting Fig. 2), thus indicating that the Smad2-dependent pathway is activated regardless of Tgfbr2 status,

perhaps through activin signaling in the Trp53KO;Tgfbr2KO mice. The status of Smad3 was also assessed in the tumor samples (Fig. 5A). In contrast to Smad2, increased total Smad3 protein was observed in the majority of tumors from Trp53KO mice as compared with tumors from Trp53KO;Tgfbr2KO mice. This increase in total Smad3 levels corresponded to an overall increase in phospho-Smad3 levels in the Trp53KO tumors and suggests that regulation of total Smad3 levels and subsequent Smad3-dependent signaling may promote the tumors of the Trp53KO mice. Next, we analyzed the activation status of the mitogen-activated protein kinase

(MAPK) pathway, another signaling cascade Atorvastatin that can be induced by TGF-β1 stimulation (Fig. 5B). Interestingly, we found that the MAPK pathway, as measured by phospho-extracellular signal-regulated kinase (ERK)1/2, is highly activated in the Trp53KO tumors compared with tumors lacking both p53 and Tgfbr2. Furthermore, increased ERK1/2 phosphorylation is also observed in the normal liver tissue in the Trp53KO mice as compared with the normal tissue from the Trp53KO;Tgfbr2KO mice. This increase in phosphorylated ERK1/2 in Trp53KO tumors was also observed by immunohistochemistry (Fig. 6; Supporting Fig. 3). TGF-β induces the expression of a number of downstream target genes that regulate various cellular processes including proliferation, angiogenesis, and tissue remodeling.

To visualize filamentous actin in order to document cell shape, F

To visualize filamentous actin in order to document cell shape, FITC-coupled phalloidin (1 μg/mL) was applied. Lysed samples from perfused liver or HepG2 cells were transferred to sodium dodecyl sulfate / polyacrylamide gel electrophoresis (SDS-PAGE) and proteins were then blotted to nitrocellulose membranes using a semidry transfer

apparatus (GE Healthcare, Freiburg, Germany). Blots were blocked for 1 hour in 5% (w/v) bovine serum albumin (BSA) or milk powder containing 20 mmol/L Tris, pH 7.5, 150 mmol/L NaCl and 0.1% Tween BYL719 mouse 20 (TBS-T), then incubated at 4°C overnight with the respective first antibody (antibodies used: anti-phospho-EGFR-Y845,

-Y1045, Y1173, anti-β1 integrin [1:2,500], anti-phospho-Erk-1/-2, anti-Erk-1/-2, anti-rat Ntcp [K4], anti-EGFR [1:5,000], and anti-γ-tubulin 91:10,000]). Following washing with TBS-T and incubation with horseradish peroxidase-coupled MAPK Inhibitor Library purchase antimouse, or antirabbit IgG antibody (all diluted 1:10,000) at room temperature for 2 hours, the blots were washed extensively and developed using enhanced chemiluminescent detection (GE Healthcare). Blots were then analyzed densitometrically and normalized to total protein amount. Two different FLAG tags were cloned to the NH2 terminus of rat Ntcp using partially overlapping oligonucleotides. The sequence of the first primer pair was 5′-ATG GAC TAC AAG GAC GAT GAC GAT AAG AGG-3′ and 5′-CTT ATC GTC ATC GTC CTT GTA GTC CAT CCT-3′ and coded for a start codon, followed by the conventional FLAG tag (DYKDDDDK). The second primer pair was 5′-CC GCC ATG GAC TAC AAG GAC GAT GAC GAT AAG AGG-3′ and 5′-CTT ATC

GTC ATC GTC CTT GTA GTC CAT GGC GGC CT-3′, which codes for a FLAG tag with an improved new start codon (Kozak sequence). The free ends of the annealed products were complementary to the restriction sites of the endonuclease Van91I. The products were cloned into the Ntcp-enhanced green fluorescent protein (EGFP) plasmid after digestion with Van91I in-frame to the 5′-site of Ntcp.25 The accuracy of the resulting plasmids was confirmed by sequencing. HepG2 cells were cultured in Dulbecco’s modified Eagle’s medium Nutrimix F12 (Biochrom, Berlin, Germany) containing 5% fetal calf serum (PAA, Coelbe, Germany), as described.26 FLAG-Ntcp-EGFP was transfected into HepG2 cells using Lipofectamine 2000 (Life Technologies, Darmstadt, Germany) according to the manufacturer’s guidelines. Stable cell lines were established with 0.5% of geneticin as selection agent.

Further work needs to be conducted to validate these preliminary

Further work needs to be conducted to validate these preliminary results. The results of this study will contribute to a better understanding of the pathophysiology of ischemic type biliary strictures and possible treatment options. KY MAK,1 R CHIN,3 J TORRESI,3 S CUNNINGHAM,4 I ALEXANDER,4 PW ANGUS,2 CB HERATH1 1Department of Medicine, The University of Melbourne, Melbourne, Australia, 2Liver Transplant Unit, Austin Health, Melbourne, Australia, 3Department of Infectious Disease, Austin Health, Melbourne, Australia, 4The Children’s Hospital at Westmead, University of Sydney, Sydney, Australia Background: Recent

selleck chemical studies suggest that the alternate arm of the RAS consisting of ACE2, angiotensin 1-7 (Ang-1-7) and its receptor, Mas, is a potential therapeutic target in liver fibrosis.1,2,3 this website ACE2 appears

to be a negative regulator of the RAS by degrading potentially deleterious vasoconstrictor and profibrotic actions angiotensin II (Ang II) to produce Ang-(1-7), a peptide that has anti-fibrotic activity. We therefore investigated a long-term therapeutic effect of ACE2 in mice with experimental liver disease. Methods: A single injection of recombinant AAV2/8 carrying murine ACE2 (rAAV2/8-ACE2) with a liver-specific promoter was intra-peritoneally administered to mice with liver disease induced by bile duct ligation (BDL), carbon tetrachloride (CCl4) injection and methionine and choline deficient (MCD) diet feeding. The mice were sacrificed 1 week (BDL) and 6 weeks (CCl4 and MCD) after ACE2 treatment. To determine hepatic fibrosis, gene and protein expressions of collagen and pro-fibrotic mediators, and effects on Ang II signaling

pathways were analyzed. Results: Untreated mice showed extensive hepatic fibrosis at 2 weeks after BDL and 8 weeks after and CCl4 Y27632 injections and MCD diet feeding. However, ACE2 therapy for 1 week (BDL) and 6 weeks (CCl4 and MCD) significantly reduced fibrosis, as reflected by marked reductions in liver hydroxyproline content and picrosirius red staining compared to controls. In both models gene expression of collagen 1 (COL1A1), alpha-smooth muscle actin (α-SMA), connective tissue growth factor (CTGF) and transforming growth factor beta (TGF-β) were significantly down-regulated in ACE2 treated mice. These changes were accompanied by increases in hepatic levels of the antifibrotic peptide Ang-(1-7) and reduced Ang II levels, with associated reductions in membrane translocation of the cytoplasmic p67phox NADPH oxidase subunit and activation of p38 MAP kinase. Conclusion: We conclude that rAAV2/8-ACE2 reduces fibrosis by changing the intrahepatic balance of Ang II and Ang-(1-7) production in the liver and may be an effective therapeutic option for the treatment of hepatic fibrosis. 1. Grace JA., et al., Update on new aspects of the renin-angiotensin system in liver disease: clinical implications and new therapeutic options.