In the present study, we investigated the molecular mechanisms by which NS4B targets RIG-I–induced and STING-mediated IFN-β production signaling. IFN-β promoter reporter assay showed that IFN-β promoter activation induced by RIG-I or Cardif was significantly suppressed by both NS4B and NS3/4A, whereas STING-induced IFN-β activation was suppressed by NS4B but not by NS3/4A, suggesting that NS4B had a distinct point of interaction. Immunostaining showed that STING colocalized with NS4B in the endoplasmic reticulum.

Immunoprecipitation and bimolecular fluorescence complementation (BiFC) assays demonstrated that NS4B specifically bound STING. Intriguingly, NS4B expression check details blocked the protein interaction between STING and Cardif, which is required for robust IFN-β activation. NS4B truncation assays showed that its N terminus, containing the STING homology domain, was necessary for the suppression of IFN-β promoter activation. NS4B suppressed residual IFN-β activation by an NS3/4A-cleaved Cardif (Cardif1-508), suggesting that NS3/4A and NS4B may cooperate in the blockade of IFN-β production. Conclusion:

NS4B suppresses RIG-I–mediated IFN-β production signaling through a direct protein interaction with STING. Disruption of that interaction may restore cellular antiviral responses and may constitute a novel therapeutic strategy for the eradication of HCV. (HEPATOLOGY 2013) Type I interferon (IFN) plays a central role in eliminating hepatitis C virus (HCV) both under physiological conditions and when used as a therapeutic intervention.1-3 In experimental acute-resolving HCV infection in chimpanzees, MCE numerous Cetuximab IFN-related genes are expressed during clinical course of infection.4 Viruses are recognized by cellular innate

immune receptors, such as toll-like receptors, and a family of RIG-I–like receptors, such as retinoic-acid-inducible gene I (RIG-I) and melanoma-differentiation-associated gene 5 (MDA-5); host antiviral responses are then activated, resulting in the production of cytokines such as type I and type III IFNs.5 RIG-I is activated through recognition of short double-strand RNA (dsRNA) or triphosphate at the 5′ end of dsRNA as pathogen-associated molecular patterns,6, 7 forming a homo-oligomer that binds with the caspase recruitment domain (CARD) of Cardif (also known as MAVS, VISA, or IPS-1).8-11 Cardif subsequently recruits TANK binding kinase 1 (TBK1) and IκB kinase ϵ (IKKϵ) kinases, which catalyze phosphorylation and activation of IFN regulatory factor-3 (IRF-3).12 Activation of TBK1 and IKKϵ results in the phosphorylation of IRF-3 or IRF-7, translocation to the nucleus, and induction of IFN-β mRNA transcription. Several HCV proteins can block host cellular antiviral responses. HCV core protein blocks IFN signaling by interacting with signal transducer and activator of transcription protein-1 (STAT1).

CIMT was graded 0: normal; 1: <1 mm wall thickening without CP; 2: moderate CP (≥1-≤2.5 mm), MK0683 price 3: CP>2.5 mm). Progression was defined as transition to the next, higher class. Steatosis and fibrosis were assessed by the Fatty Liver Index (FLI, NAFLD present when >60), and by FibroTest (FT). Results.2169 patients were enrolled: 56% males, 52 year-old, BMI 25.4 kg/m2; mean FRS 12±9%, mean CIMT 0.62±0.14 mm; 40.5% had CP and 24% had a FLI60. Pts with NAFLD had higher CIMT (0.64±0.15 vs.0.61 ±0.14 mm, p=0.001), higher prevalence of CP (30% vs.24%, p=0.005) and higher

FRS (17±9% vs.10±8%, p<0.001). FLI was associated with baseline CIMT (p=0.002) and FLI≥60 with CP at baseline (OR=1.27, p=0.04), both independent of age, sex, smoking, diabetes and hypertension. The median f/u was 8 yrs, 6.4 yrs in NAFLD pts and 8.5 yrs in non-NAFLD pts (p<0.001). During f/u, CIMT Anti-infection Compound Library nmr increased

from 0.62 to 0.65 mm, p<0.001, prevalence of CP increased from 41% to 58%, p<0.001 while 38% of pts developed CP. NAFLD at baseline was associated with the progression and occurrence of CP independent of age, sex or the FRS (HR 1.30 and 1.34, respectively both p<0.01). Among non-NAFLD pts at baseline, those who developed NAFLD during f/u had a larger increase in CIMT than those who stayed NAFLD-free.455 pts were evaluated by FT, they were not different from the 1714 untested pts for age, BMI, CIMT and CP prevalence.2% of these patients had a FT>0.48 compatible with bridging fibrosis. Unexpectedly, bridging fibrosis was associated with the presence of CP at baseline and with progression of CP at f/u (HR 3.83, p<0.01), both independent of FRS and FLI>60. Conclusion: In patients at high CV risk, NAFLD and in particular bridging fibrosis contribute to early atherosclerosis and progression thereof, independent of traditional CV risk factors. Disclosures: Pascal Lebray – Grant/Research Support: Schering Plough; Speaking and Teaching:

Janssen, MSD, Gilead Mona Munteanu – Employment: Biopredictive Thierry Poynard – Advisory Committees or Review Panels: Merck; Grant/Research Support: BMS, Gilead; Stock Shareholder: Biopredictive Vlad Ratziu – Advisory Committees or Review Panels: GalMed, MCE Abbott, Genfit, Enterome, Gilead; Consulting: Astellas, Axcan, Pfizer, Sanofi-Synthelabo, Genentech, Nycomed The following people have nothing to disclose: Raluca Pais, Philippe Giral, Hugo Perazzo, Jean-Francois Khan, Larysa Fedchuk, David Rosenbaum Background & Aims: Epicardial adipose tissue (EAT) has been implicated in the pathogenesis of coronary atherosclerosis, and its measurement during echocardiography proposed as a new index of cardiac and visceral adiposity. EAT was found increased in patients with metabolic syndrome, of which nonalcoholic fatty liver disease (NAFLD) represents the hepatic manifestation.

Hopefully, the initiation FK506 of the World Health Organization International Clinical Trials Registry Platform will facilitate such

assessments for future trials.41, 42 Another limitation in this review was insufficient reporting. Investigators of future trials are therefore well advised to adhere to the Consolidated Standards for Reporting of Trials in order to improve the quality of trial reports.43 These potential limitations and concerns may lower our confidence in the estimates of intervention effect. However, in our meta-analysis for SVR there is no apparent heterogeneity (I2 = 0%), and the direction of the treatment effect is the same across all included trials. Further research is unlikely to change our confidence in the estimate of the effect. It is a common misconception that large RCTs are generally more reliable than meta-analyses. The reason this misconception has prevailed is due to a number of highly

cited papers that compared high-quality large trials with collections of low-quality small trials (an unfair comparison). In empirical studies where high-quality large trials are compared with a collection of high-quality small trials, the results from the two are typically nondiscrepant. In the case of the IDEAL trial,3 the results still show an effect—albeit small—in favor of peginterferon alpha-2a. There are many examples of large trials that underestimate the treatment effect simply by chance. Current evidence suggests that peginterferon alpha-2a is significantly superior to find more peginterferon alfa-2b regarding benefits (SVR, which is clearance of the virus from the blood). However, there is insufficient evidence to detect any differences regarding harms (mortality and adverse events). Future trials must further the correlation between achieving SVR and clinically relevant outcomes such as risk of cirrhosis, hepatocellular carcinoma, and mortality. MCE公司 We thank the patients and investigators who participated

in the included trials, with special thanks to the investigators who responded to our inquiries. We also thank our colleagues Dimitrinka Nikolova and Sarah Louise Klingenberg. “
“Dorrell C, Erker L, Schug J, Kopp JL, Canaday PS, Fox AJ, et al. Prospective isolation of a bipotential clonogenic liver progenitor cell in adult mice. Genes Dev 2011;25:1193-1203. (Reprinted with permission.) The molecular identification of adult hepatic stem/progenitor cells has been hampered by the lack of truly specific markers. To isolate putative adult liver progenitor cells, we used cell surface-marking antibodies, including MIC1-1C3, to isolate subpopulations of liver cells from normal adult mice or those undergoing an oval cell response and tested their capacity to form bilineage colonies in vitro.

Hopefully, the initiation ABT-263 clinical trial of the World Health Organization International Clinical Trials Registry Platform will facilitate such

assessments for future trials.41, 42 Another limitation in this review was insufficient reporting. Investigators of future trials are therefore well advised to adhere to the Consolidated Standards for Reporting of Trials in order to improve the quality of trial reports.43 These potential limitations and concerns may lower our confidence in the estimates of intervention effect. However, in our meta-analysis for SVR there is no apparent heterogeneity (I2 = 0%), and the direction of the treatment effect is the same across all included trials. Further research is unlikely to change our confidence in the estimate of the effect. It is a common misconception that large RCTs are generally more reliable than meta-analyses. The reason this misconception has prevailed is due to a number of highly

cited papers that compared high-quality large trials with collections of low-quality small trials (an unfair comparison). In empirical studies where high-quality large trials are compared with a collection of high-quality small trials, the results from the two are typically nondiscrepant. In the case of the IDEAL trial,3 the results still show an effect—albeit small—in favor of peginterferon alpha-2a. There are many examples of large trials that underestimate the treatment effect simply by chance. Current evidence suggests that peginterferon alpha-2a is significantly superior to Cisplatin cell line peginterferon alfa-2b regarding benefits (SVR, which is clearance of the virus from the blood). However, there is insufficient evidence to detect any differences regarding harms (mortality and adverse events). Future trials must further the correlation between achieving SVR and clinically relevant outcomes such as risk of cirrhosis, hepatocellular carcinoma, and mortality. medchemexpress We thank the patients and investigators who participated

in the included trials, with special thanks to the investigators who responded to our inquiries. We also thank our colleagues Dimitrinka Nikolova and Sarah Louise Klingenberg. “
“Dorrell C, Erker L, Schug J, Kopp JL, Canaday PS, Fox AJ, et al. Prospective isolation of a bipotential clonogenic liver progenitor cell in adult mice. Genes Dev 2011;25:1193-1203. (Reprinted with permission.) The molecular identification of adult hepatic stem/progenitor cells has been hampered by the lack of truly specific markers. To isolate putative adult liver progenitor cells, we used cell surface-marking antibodies, including MIC1-1C3, to isolate subpopulations of liver cells from normal adult mice or those undergoing an oval cell response and tested their capacity to form bilineage colonies in vitro.

Results: We found that TG (P=0.004) and LDL-C (P=0.02) but not HDL-C GWA SNP sets were enriched in NAFLD. We identified 58 pathways that were enriched in lipid GWAS data. Three of these were also enriched in the NAFLD GWAS (N=7,

126) and one, FXR/RXR activation, also showed significant enrichment in the second independent NAFLD GWAS (N=3, 124). None of the three Doxorubicin in vitro original NAFLD enriched pathways were enriched for associations in control publically available GWAS analyses of diastolic and systolic blood pressure (N=275, 000), body mass index (N=249, 796), and waist to hip ratio (N=77, 167) suggesting that the enrichment was specific to NAFLD. Genes associated with NAFLD (P<0.05) in FXR/RXR activation fell into three functional categories: (1)VLDL Assembly: MTTP, APOB, APOC3, (2) Nuclear Related Processes: PPAR-a, HNF1 a, NR0B2/SHP and (3) Hepatic Transport: MRP2, AE2, ABCG8, ABCG5, OAT2. Conclusions: Using a novel approach, we found that human genetic variation in or near genes involved in FXR/RXR activation affects both blood lipids and NAFLD in humans. These results suggest that genes that play a role in lipoprotein assembly, nuclear receptor biology, and hepatic transport when altered may affect NAFLD and thus could provide possible therapeutic targets for NAFLD prevention or treatment. Disclosures:

The following people have nothing to disclose: Yindra M. Puentes, Corey Opaganib cost C. Powell, Laura M. Yerges-Armstrong, Mary F. Feitosa, Lawrence F. Bielak, Albert V. Smith, Tamara B. Harris, Jiankang Liu, Solomon K. Musani, Ingrid B. Borecki, Patricia A. Peyser, Elizabeth K. Speliotes Increased circulating soluble CD36 (sCD36), a cell-free form of fatty acid translocase CD36, clusters with insulin resistance and surrogate markers of fatty liver in population studies but no evidence exists on its

relationship with hepatic fat content. The aim of the present study was to elucidate whether circulating sCD36 is linked to the amount of lipids within the liver in nonalcoholic fatty liver disease (NAFLD) and chronic hepatitis C (CHC) patients. This study comprised an overall population of 399 patients (227 with NAFLD, 87 with CHC, and 85 with histologically normal liver [NL]) who underwent a liver MCE公司 biopsy either by a percutaneous route for diagnostic purposes or during programmed abdominal surgery for gastroplasty or cholecystectomy. Steatosis was graded by Kleiner histological scoring system. Serum sCD36 levels and hepatic CD36 expression were assessed by immunoassay and immunohistochemistry, respectively. In NAFLD patients, serum sCD36 levels were significantly higher in those with simple steatosis than in NL subjects (361.4 ± 286.4 versus 173.9 ± 137.4 pg/mL, respectively; P < 0.001) but not in patients with steatohepatitis (229.

Our results suggest that the consumption of several prey categories fluctuates significantly year to year. Few data are available to indicate abundance of the main prey categories, although fishery statistical

data from ICES subarea IX (west of the Iberian Peninsula) suggest that ommastrephid (virtually all of which will be Illex coindetii and Todaropsis eblanae, Pierce et al. 2010b) abundance has fluctuated widely. Landings in the early 1990s were low, as little as 250 tons LBH589 in 1993, before rising to a peak of almost 3,000 tons in 1997 before declining again reach slightly over 300 tons in 2007. A similar trend was seen in Bay of Biscay waters (ICES 2000, 2011). Our dietary data are clearly inadequate to test whether diet has tracked prey abundance, selleck but there was evidence of a decline in the numerical importance of Illex and Todaropsis in pilot whale diet during approximately 2000 to 2005. The higher importance of octopus in the diet of pilot whales found in the present study (and by Spitz et al. 2011) compared to most previous studies probably reflects a latitudinal trend, with squids (mainly ommastrephids) dominating the diet at higher latitudes

while octopods are more important at lower latitudes. These differences could relate to differences in prey availability, but there are no relevant abundance estimates for these cephalopod groups and this hypothesis is not presently testable. Improving our knowledge of the factors affecting the diet of deep divers such as pilot whales could help us to understand the trophic links within these systems and also the relationships between oceanic and shelf waters that this predator seems to be able to exploit simultaneously.

It would be interesting to understand why the whales appear to take mostly prey species of relatively low energy density. Few data exist on the calorific values of oceanic cephalopods although some figures are available for MCE neritic species. For example, Spitz et al. (2011) gave values of 4.7 kJ/g for E. cirrhosa and 4.4 kJ/g for squid of the family Ommastrephidae (only Illex coindetti and Todaropsis eblanae were analyzed). These values are similar to those for fish of the family Gadidae but are quite low when compared with the energetic content of some other fish such as clupeids and some myctophids. In principle, diet selection is expected to reflect a trade-off between calorific content of the prey and the energetic cost of capturing them, suggesting that prey species such as Eledone cirrhosa may be particularly abundant and/or easy to capture. However, it is also true that not all biases can be accounted for when inferring the diet of a species by the analysis of the stomach contents of stranded individuals, e.g.

Changes in the S/L proteins ratio may lead to the retention of surface proteins within the hepatocytes, although this event may not affect virion secretion.18-23 Several recent studies suggest that circulating HBsAg levels might reflect intrahepatic HBV activity and that their decline under treatment might predict a response to antiviral therapies in chronic hepatitis B (CHB) patients.24-27 HBsAg titer has been proposed as a surrogate marker for HBV covalently closed circular DNA (cccDNA),24, 25, 27 the intranuclear replicative intermediate that constitutes the reservoir responsible

for viral persistence. However, studies evaluating HBsAg titers have ZD1839 purchase not considered the possible emergence of preS/S HBV mutants as dominant infecting viral populations, and in particular, they have not

taken into consideration the possible influence of preS/S gene variability on circulating levels of HBsAg. The aims of this study were to investigate whether infection with preS/S variants may influence the amounts of circulating HBsAg and HBV DNA in CHB patients and to perform a phenotypic characterization of naturally occurring preS/S variant isolates. aa, amino acid; anti-HBe, antibody to hepatitis B e antigen; BCP, basal core Selleck PCI-32765 promoter; cccDNA, covalently closed circular DNA; CHB, chronic hepatitis B; ER, endoplasmic reticulum; HBeAg, hepatitis e antigen; HBsAg, hepatitis B surface antigen; HBV, hepatitis B virus; L, large; M, medium; mt, mutant; mRNA, messenger RNA; PC, precore; PCR, polymerase chain reaction; S, small; WT, wild-type. We studied medchemexpress serum samples collected

prior to any antiviral treatment from 40 patients with HBV-related chronic liver disease (26 men and 14 women; mean age, 43 ± 14.6 years), consecutively admitted to the outpatient service of the Liver Unit of the Messina University Hospital (Italy) in 2008. All of the patients were Italian and were infected with HBV genotype D (n = 36 patients) or A (n = 4). Eleven of the patients were HBV e antigen (HBeAg)-positive, and 29 were positive for the corresponding antibody (anti-HBe). Thirteen patients had cirrhosis and 27 had CHB. The diagnosis was performed through needle liver biopsy in 17 cases and through clinical/biochemical/ultrasonographic evaluation in 23 cases (Table 1). All patients were negative for hepatitis delta virus, hepatitis C virus, and human immunodeficiency virus serum markers. The study protocol was performed according to the principles of the Declaration of Helsinki, and written informed consent was obtained from all patients. Serum HBV DNA was quantified using the COBAS AmpliPrep-COBAS TaqMan HBV test (Roche Diagnostics, Monza, Italy) with a lower limit of detection of 12 IU/mL. HBsAg was quantified on the same sera using the ARCHITECT HBsAg Assay (Abbott; Abbott Laboratories, Chicago, IL) according to the manufacturer’s instructions.

Serial dilutions of HCVcc (H77/JFH genotype 1a/2a chimera) inhibited anti-CD3/CD28–stimulated IL-2 production in a dose-dependent manner (Fig. 2). The HuT 78 T cell line secretes IL-2 following stimulation with the phorbol ester PMA. We used this model system to investigate the mechanism of HCV E2–mediated effects on reduced IL-2 production. HCV E2 significantly reduced PMA-stimulated Hut 78 cell IL-2 release compared with untreated or recombinant core (C22 or C33)-treated cells (Fig. 3A). To confirm that this effect of HCV E2 was CD81-mediated, we confirmed that the BP could reverse the effect (Fig. 3B).

Previous reports have demonstrated that CD81 is costimulatory for IL-2 production,23 consistent with our data showing that anti-CD81 cross-linking cotemporaneously with an anti-CD3 stimulus promoted IL-2 production. However, ligation of CD81 with HCV E2 alone or soluble anti-CD81 prior to T cell stimulation Apoptosis antagonist inhibited IL-2 secretion (Supporting Information Fig. 5). To ascertain whether this inhibition was at a transcriptional level, we quantified IL-2 messenger RNA (mRNA) levels using real-time PCR (Fig. 3C). IL-2 mRNA levels in PMA-stimulated cells increased 126-fold over resting cells (P = 0.02)

but this was not inhibited by HCV E2. Immunofluorescent analysis revealed cytosolic IL-2 protein in HCV E2 pretreated cells stimulated with selleck chemical PMA that was not released externally (Supporting Information Figs. 6 and 7). To investigate whether this inhibition of secretion was IL-2–specific or associated with general targeting of the secretory machinery, we examined HCV E2 effects on secretion of other cytokines in PMA-treated HuT 78 cells (Fig. 4) and anti-CD3/anti-CD28–stimulated PBMCs (Supporting Information Fig. 8). HCV E2 inhibited the secretion of interferon-γ (IFNγ), tumor necrosis factor-α (TNFα), and IL-10 from both activated HuT 78 cells and PBMCs (Fig. 4A-C, Supporting Information Fig. 8), suggesting that E2 targets a secretory

process. In contrast, E2 had minimal effect on IFNγ mRNA levels in HuT 78 cells (Fig. 4D), although there was a modest decrease in both IL-2 and IFNγ mRNA in PBMCs pretreated with E2 (Supporting Information Fig. 8). Treatment of HuT 78 cells and PBMCs with E2 prior medchemexpress to activation attenuated stimulated levels of both TNFα and IL-10 mRNA (Fig. 4E-F, Supporting Information Fig. 8), suggesting that HCV E2 can target transcriptional activation of these cytokines in T cells. Overall, the data demonstrate that HCV E2 targets the T cell secretory machinery and can inhibit secretion of IL-2 and IFNγ, cytokines that are normally secreted directionally through the centrosome.24 We have reported previously that PKCβ is necessary for IL-2 export from PMA-stimulated HuT 78 cells.16 In resting HuT 78 cells, PKCβ displays a cytosolic distribution (Fig. 5A); however, after incubation with HCV E2, PKCβ localized to lipid rafts, colocalizing with GM-1 (Fig. 5B,C).

GeneSpring 6.2 (Silicon Genetics,

Inc., Redwood City, see more CA) was then used to evaluate the data obtained using CodeLink Expression Scanning Software. Total RNA was isolated from cell samples using the RNeasy mini kit (Qiagen). One microgram of purified, DNase-treated total RNA was used to synthesize biotinylated cRNA target preparation using the CodeLink™ Expression Assay Reagent Kit (Amersham) according to manufacturer’s instructions. Ten micrograms of fragmented cRNA was applied to the Uniset Mouse I Expression Bioarray, hybridized for 18 hours, and the arrays processed according to manufacturer’s instructions using Streptavidin-Alexa647 detection reagents. Slides were scanned using a GenePix 4000B scanner (Axon) and the images were analyzed with CodeLink™ Expression Analysis Software. To investigate the role played by Hex in hepatocyte lineage commitment during EB differentiation, we induced endoderm formation from wild-type (Hex+/+), heterozygous (Hex+/−), or Hex-deficient (Hex−/−) ESCs15 using the serum induction protocol that we have described.22 Consistent with our earlier findings, the hepatocyte genes Alb and Afp were both expressed in EBs generated from the Hex+/+ and Hex+/− ESCs. The levels of expression of both genes

were markedly reduced in the PLX-4720 solubility dmso Hex−/− EBs (Fig. 1A). These findings are in line with those from studies on the early embryo, demonstrating that Hex is required for development of the liver in vivo.13, 15 To further evaluate the role of Hex in hepatic specification in the ESC/EB model, we used an ESC line (AINV18)

that enables the regulated expression of a given gene under the control of a tet-inducible promoter. Using this system, we generated ESCs in which Hex expression was induced by the addition of the tetracycline analogue Dox (tet-Hex ESCs). Hex expression was induced in the cells by the addition of Dox (1 μg/mL) to the EB cultures either from days 6–10 or from days 6–22 of differentiation. Quantitative PCR analyses revealed that induction of Hex between days 6 and 10 of culture resulted in a significant up-regulation of Afp and Alb expression compared with the uninduced cultures (Fig. 1B,C). These levels of expression at day 14 represent 2.6% and MCE 2.5% of the expression found in the fetal liver, respectively. By contrast, when Hex was continuously expressed from day 6 to day 22, levels of Alb and Afp mRNA were diminished on day 22 (Fig. 1B,C), suggesting that prolonged Hex expression may disrupt hepatic differentiation or shift the tissue into another fate. Immunostaining showed the presence of clusters of Alb+ cells within the Hex-induced day 14 EBs (Fig. 1D). Hex induction also resulted in enhanced secretion of both Alb and transferrin by the EB-derived cells at day 14 of culture (Fig. 2A,B). These levels of secretion were 7.6% and 5.0% of that of day 14 fetal liver cells, respectively.

Expression analysis revealed that UDCA-LPE exerted profound anti-inflammatory properties in HFD and MCD diet-induced liver injury. Expression of monocyte chemoattractant protein-1 (MCP-1) was up-regulated 3.8-fold, vascular cell adhesion molecule-1 (VCAM-1), was increased 4.6-fold, and TNF-α was elevated 14-fold in HFD mice, Fluorouracil in vivo whereas all three genes were down-regulated nearly to normalization in HFD mice treated with UDCA-LPE (Fig. 5A,C,E). Steatohepatitis due to the MCD diet was accompanied by even higher expression levels: nine-fold for MCP1, 22.3-fold for VCAM1, and 22.6-fold for TNF-α (Fig. 5B,D,F). Nevertheless, administration of UDCA-LPE was capable of markedly decreasing the expression of these proinflammatory

genes by 50%-75% (Fig. 5B,D,F). These results were further confirmed on the protein level for MCP-1 in both mouse models (Supporting Fig. 3). Increased cellular content of the potent lipid mediator LPC has been implicated in different inflammatory diseases. Analysis of hepatic phospholipid composition in lipid extracts showed an abundance of proinflammatory LPC in both HFD and MCD mice with up to two- to five-fold increase

in LPC concentrations, respectively (Fig. 6A,B). In contrast, lipid extracts of liver tissue of HFD and MCD mice administered with UDCA-LPE showed a pronounced decrease in intrahepatic LPC pools down to baseline levels in HFD mice as well as a reduction of LPC by INK 128 order almost one-third in mice on the MCD diet (Fig. 6A,B). In addition to proinflammatory LPC, we studied lipid peroxidation in MCD mice as a consequence of bundant reactive oxygen species (ROS) formation in fatty livers. In contrast to HFD mice, which did not display increased lipid peroxidation (data not shown), our results showed a marked rise in lipid hydroperoxide concentrations by nearly 10-fold in MCD-induced NASH. UDCA-LPE treatment

strongly inhibited the generation of this proinflammatory lipid intermediate, with a decrease in lipid hydroperoxides of 73% (Fig. 6C). NAFLD is characterized by changes in hepatic lipid homeostasis and fatty acid metabolism. Therefore, medchemexpress we aimed to study the expression of genes involved in de novo lipogenesis, triglyceride synthesis, and desaturation of fatty acids. As expected for the HFD model, we found enhanced de novo lipogenesis with up-regulation of acetyl-CoA carboxylase 1 (ACC1), fatty acid synthetase (FASN), and their transcriptional regulator sterol regulatory element binding protein 1c (SREBP1c) (Fig. 7A). In contrast, HFD mice treated with UDCA-LPE showed a decrease in ACC1 and SREBP1c transcripts to levels of control mice, as well as a marked reduction in FASN expression (Fig. 7A) and protein levels (Supporting Fig. 4) of almost 50%. As for the MCD diet, which causes an impairment of de novo lipogenesis,22 UDCA-LPE administration resulted in partial restoration of expression levels of lipogenic genes (Supporting Fig. 5).