The TLR4 agonist, LPS and the TLR2/Dectin-1 agonist, Zymosan both potently induced G-CSF secretion by PBMCs, which was significantly suppressed by co-incubation with IFN-α (data not shown). As we found that PBMCs isolated from patients on IFN-α/ribavirin therapy did MK-8669 mw not secrete high levels of G-CSF (Fig. 1b), we wished to determine whether PBMCs isolated from these individuals could produce G-CSF in response to in vitro stimulation

with a TLR7/8 agonist that effectively drives G-CSF secretion by PBMCs (Fig. 3a). Therefore, we stimulated PBMCs isolated from HCV-infected patients receiving IFN-α/ribavirin therapy at week 24 of their treatment with CL097 and found that they secreted high levels of G-CSF in response to this stimulation (Fig. 4). Interferon-α has potent anti-viral activity and is the mainstay of anti-viral therapy for patients with chronic HCV infection. However, IFN-α has

significant toxic effects on the hematopoietic system. IFN-induced neutropenia frequently causes dose reduction or Adriamycin manufacturer treatment discontinuation. Bone marrow suppression contributes to the development of IFN-induced cytopenias.7 However, the effect of IFN-α on the expression of the hematopoietic growth factors that affect the development and efflux of neutrophils from bone marrow has not been studied in detail. G-CSF regulates neutrophil development. Mice lacking G-CSF have chronic neutropenia, granulocyte and macrophage progenitor cell deficiency and impaired neutrophil mobilization.17 Therefore, we investigated the effects of IFN-α on G-CSF production by PBMCs both in vitro and ex vivo from patients who had received therapeutic IFN-α to treat chronic HCV infection. We found that PBMCs isolated from patients on IFN-α/ribavirin therapy gradually lose the ability to produce G-CSF over the course of the treatment (Fig. 1b). The decline in the ability of patients’ PBMCs to produce G-CSF in culture paralleled the reduction in ANC over the course of IFN-α treatment, suggesting that suppressed G-CSF production by PBMCs may contribute to

click here IFN-α-induced neutropenia. Reduced G-CSF production by PBMCs may explain the suppressive effect of IFN-α on progenitor cell proliferation in bone marrow.7 The precise mechanism by which IFN-α exerts its suppressive effect on G-CSF production is unclear, in part because the mechanisms underlying the regulation of G-CSF production in vivo remain poorly defined.18 However, our finding that a TLR7/8 agonist induces G-CSF production in human monocytes suggests that NFκB has a role in the regulation of its expression. This is further confirmed by the recent finding that serum amyloid A (SAA) induces G-CSF production in mouse macrophages via a TLR2 dependent pathway.19 G-CSF stimulates angiogenesis and tumor growth.

Recently, subjects with low vitamin D levels are associated with metabolic syndrome and diabetes.

However, associations between low vitamin levels, and ultra-sonographically diagnosed http://www.selleckchem.com/products/MG132.html NAFLD and advanced fibrosis in patients with NAFLD have not been completely established. The aim of this study was to characterize the relationship between vitamin D levels and NAFLD in the large, national, general population. Methods: The Third National Health and Nutrition Examination Survey (NHANES) from 1988 to 1994 were utilized in this study. A total of 11,808 participants without known liver disease were finally selected and analyzed. NAFLD was defined as ultrasonography diagnosed hepatic steatosis without other known liver diseases. The presence and absence of advanced fibrosis in NAFLD was determined by the NAFLD fibrosis score. Results: The prevalence of ultrasonography diagnosed NAFLD was 22.9%. NAFLD was significantly inversely associated with vitamin levels after adjusted for age and sex [odds ratio (OR) 0.85 95% confidence interval (CI) 0.75-0.96]. The age, sex-adjusted prevalence of NAFLD decreased steadily with increasing levels of vitamin D [OR 0.53 95% LY2109761 mw CI 0.40-0.70, lowest quintile vs. highest quintile, p for trend <0.001]. Multivariate regression analysis after adjustment for known

risk factors showed that NAFLD was statistically significantly inversely associated with vitamin levels (>20 ng/ml) [OR 0.79, 95% CI, 0.65-0.95] and the grade of vitamin levels in a dose-dependent manner [OR 0.76, 95% CI, 0.58-1.00 in 5th quintile vs. lowest quintile, p for trend=0.001]. With regards to advanced fibrosis, age, sex-adjusted advanced fibrosis in patient with NAFLD decreased steadily with increasing levels of vitamin D [OR 0.23 95% CI 0.08-0.66, highest tertile vs. lowest tertile, p for trend <0.001]. Multivariate

regression analysis after adjustment for known risk factors showed that NAFLD was statistically significantly inversely associated with the grade of vitamin levels in a dose-dependent manner [OR 0.17, 95% CI, 0.24-0.74 in highest tertile vs. lowest tertile]. Conclusions: Serum vitamin D, even in the range of normal levels, was found to be inversely related to NAFLD in a dose-dependent manner. Vitamin D is inversely selleck associated with advanced fibrosis in patients with NAFLD independently of known metabolic risk factors. Vitamin D might have protective effect for NAFLD and advanced fibrosis. Disclosures: The following people have nothing to disclose: Donghee Kim, Won Kim, Hwa Jung Kim Background: There is an urgent need to validate non-invasive markers to quantify fibrosis because of invasive nature and complications of Liver Biopsy. Aim: To compare various non invasive markers available for liver fibrosis including Lok score, Fibro Index(FI)score, King score, Fibro Q score, AST to Platelet Ratio Index(APRI), FIB 4 score and AST to ALT ratio(AAR)with Ishak stage(IS).

Mounting evidence implicates that Mesenchymal Stem Cells (MSCs) have properties of low immunogenicity, immunomodulation and anti-inflammatory in vitro and in vivo, especially regulate T-cell responses. Therefore, we transplanted MSCs into an experimental model of IBD to investigate their potential therapeutic effects in vivo. Methods: A rectal enema of trinitrobenzene sulfonic acid (TNBS) (100 mg/kg body weight) was applied

to female BALB/c mice. Bone marrow mesenchymal stem cells (BMSCs) were derived from male green fluorescent protein (GFP) transgenic mice and were transplanted intravenously into animal models after onset of disease. Clinical activity scores, histological changes were evaluated. GFP and Sex determining Region Cilomilast mw Y gene (SRY) expression were used for cell tracking. Ki67 positive cells and Lgr5-expressing cells were detected to observe proliferative activity.

Inflammatory response was determined by measuring levels of different inflammatory mediators in colon and serum. The inflammatory cytokines include tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ),interleukin-2(IL-2),IL-6,IL-17,IL-4,IL-10,transforming growth factor(TGF-β).Master regulators of Th1 cell (T-box Selleckchem ACP-196 expressed in T cells,T-bet),Th17 cell(retinoid related orphan receptor gammat,RORγt),Th2 cell (GATA family of transcription factors 3,GATA3) and regulatory T cell selleck screening library (forkhead box P3,Foxp3) were also detected. Results: Systemic infusion of GFP-BMSCs ameliorated the clinical and histopathologic severity of colitis, including body weight loss, diarrhea, inflammation and increasing survival (P < 0.05). Cell tracking

study showed that MSCs homed to the injured colon. MSCs promoted proliferation of intestinal epithelial cells and differentiation of intestinal stem cell (P < 0.01). This therapeutic effect was mainly mediated by down-regulating both Th1-Th17 driven autoimmune and inflammatory responses (IL-2, TNF-α, IFN-γ, T-bet; IL-6, IL-17, RORγt), while by up-regulating Th2 activities (IL-4, IL-10, GATA-3) (P < 0.05). MSCs also induced activated CD4 + CD25 + Foxp3+ regulatory T cells (TGF-β, IL-10, Foxp3) with suppressive capacity on Th1-Th17 effecter responses and promoted Th2 differentiation in vivo (P < 0.05). Conclusion: MSCs emerge as key regulators of immune and inflammatory responses and as an attractive candidate for a cell-based therapy for IBD. Key Word(s): 1. MSCs; 2. IBD; 3. transplantation; 4. immunomodulation; Presenting Author: XUESONG YANG Additional Authors: LIPING ZHANG, WEI FU, AIYING WANG Corresponding Author: XUESONG YANG Affiliations: No Objective: To study the clinical characters and endoscopic manifestation of ulcerative colitis associated colorectal cancer (UC-CRC).

Mounting evidence implicates that Mesenchymal Stem Cells (MSCs) have properties of low immunogenicity, immunomodulation and anti-inflammatory in vitro and in vivo, especially regulate T-cell responses. Therefore, we transplanted MSCs into an experimental model of IBD to investigate their potential therapeutic effects in vivo. Methods: A rectal enema of trinitrobenzene sulfonic acid (TNBS) (100 mg/kg body weight) was applied

to female BALB/c mice. Bone marrow mesenchymal stem cells (BMSCs) were derived from male green fluorescent protein (GFP) transgenic mice and were transplanted intravenously into animal models after onset of disease. Clinical activity scores, histological changes were evaluated. GFP and Sex determining Region LY2606368 Y gene (SRY) expression were used for cell tracking. Ki67 positive cells and Lgr5-expressing cells were detected to observe proliferative activity.

Inflammatory response was determined by measuring levels of different inflammatory mediators in colon and serum. The inflammatory cytokines include tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ),interleukin-2(IL-2),IL-6,IL-17,IL-4,IL-10,transforming growth factor(TGF-β).Master regulators of Th1 cell (T-box Selleck BGB324 expressed in T cells,T-bet),Th17 cell(retinoid related orphan receptor gammat,RORγt),Th2 cell (GATA family of transcription factors 3,GATA3) and regulatory T cell click here (forkhead box P3,Foxp3) were also detected. Results: Systemic infusion of GFP-BMSCs ameliorated the clinical and histopathologic severity of colitis, including body weight loss, diarrhea, inflammation and increasing survival (P < 0.05). Cell tracking

study showed that MSCs homed to the injured colon. MSCs promoted proliferation of intestinal epithelial cells and differentiation of intestinal stem cell (P < 0.01). This therapeutic effect was mainly mediated by down-regulating both Th1-Th17 driven autoimmune and inflammatory responses (IL-2, TNF-α, IFN-γ, T-bet; IL-6, IL-17, RORγt), while by up-regulating Th2 activities (IL-4, IL-10, GATA-3) (P < 0.05). MSCs also induced activated CD4 + CD25 + Foxp3+ regulatory T cells (TGF-β, IL-10, Foxp3) with suppressive capacity on Th1-Th17 effecter responses and promoted Th2 differentiation in vivo (P < 0.05). Conclusion: MSCs emerge as key regulators of immune and inflammatory responses and as an attractive candidate for a cell-based therapy for IBD. Key Word(s): 1. MSCs; 2. IBD; 3. transplantation; 4. immunomodulation; Presenting Author: XUESONG YANG Additional Authors: LIPING ZHANG, WEI FU, AIYING WANG Corresponding Author: XUESONG YANG Affiliations: No Objective: To study the clinical characters and endoscopic manifestation of ulcerative colitis associated colorectal cancer (UC-CRC).

Using this technique, changes in regional tissue volume can be detected on the basis of the deformation field derived from the warping subject to the template image. A voxelwise estimation of the morphological differences http://www.selleckchem.com/products/atezolizumab.html between the two groups can be acquired after applying a threshold (P < .001, uncorrected) to the statistic maps. Regions with enlarged volume in the brains of blind subjects are mainly localized in the left associated visual cortex, posterior cingulated cortex, and cerebellum,

whereas volume reductions are primarily localized in the left early visual cortex. DBM is an effective method for detecting entire brain structural changes in blindness. Visual deprivation actually alters the local structural organization

during the early critical periods of neurodevelopment. Volume increases outside the occipital lobe detected with DBM may suggest compensatory adaptations. Blindness provides a rare model to investigate the effects of visual experience on the functional and structural organization of the human brain.[1] Many studies have demonstrated that the human brain can adapt to changes in the environment through functional reorganization rather than structural plastic changes. The cross-modal plasticity in functional reorganization induced by vision deprivation has also been reported in previous Tanespimycin in vivo neuroimaging. Plastic changes have been reported in the visual cortex both in the resting state and imaginary, auditory, and tactile tasks,[1-3] and in the early sensory areas of spared modalities after visual deprivation in early life.[4] Compared with functional studies, investigations on the structural reorganization of the brain in blind subjects are few and have only attracted this website research attention in recent years. Evidence from the nonhuman primate literature proves that early visual deprivation leads

to changes in the structural anatomy of the visual cortex at the microscopic level.[5], [6] With the development of the imaging technique, the identification of structural changes in the brain on magnetic resonance imaging (MRI) scans is becoming increasingly important in the study of neurological science. Using voxel-based morphometry (VBM) method,[7] previous MRI studies on blindness have reported decreased gray matter (GM) density and increased white matter (WM) density in the sensory–motor system.[8-10] Another useful tool aside from VBM for the analysis of brain morphology from MRI is the deformation-based morphometry (DBM) method which provides an unbiased automated examination of the entire brain with high regional sensitivity.[11] Unlike VBM which focuses on an analysis of differences in GM and WM, DBM can detect the differences in local structures using high-dimensional nonrigid registration. Leporé and colleagues used this technique with fast fluid registration to examine whole brain volumetric changes in both early- and late-onset blind subjects.

07). The actuarial probability of developing clinical decompensation was significantly different among the three BMI groups (log-rank 7.60, P = 0.022), being highest in obese patients, intermediate in overweight patients, and lowest in those with a normal BMI (Fig. 1B). The cumulative probability of clinical decompensation at 2 and 5 years for each BMI group was: normal weight 0% (95% confidence interval [CI] 0%-0%) and 13% (95% CI 3%-23%), respectively; overweight patients 14% (95% CI 6%-22%) and 28% (95% CI 17%-39%), respectively; obese patients 21% (95% CI 10%-32%) and 37% (95% CI 23%-50%), respectively. In a sensitivity

analysis, the increased risk of decompensation of obese patients was documented both in American patients (5-year probability: 35%; 95% CI 18%-53%) and in European patients (5-year probability: 39%; check details 95% CI 18%-61%). To evaluate whether BMI is an independent predictor of decompensation, we performed a Cox regression analysis including previously defined predictors of decompensation (HVPG, albumin, and MELD)2 and variables that could potentially act as confounders on the association (etiology and treatment). Therefore, variables introduced into the analysis were: etiology (alcoholic versus nonalcoholic); MELD score, albumin, HVPG; BMI; and treatment group (timolol or placebo). Table

2 shows the results of the uni- and multivariate Cox analysis. As shown, HVPG (per 1 mmHg increase hazard ratio, HR: 1.14 [95% CI 1.07-1.20], P < 0.0001), albumin (per 1 g/dL decrease selleck compound HR 4.54 [2.44-8.33], P < 0.0001), and BMI (per 1 unit increase HR 1.06 [1.01-1.12], P = 0.02) remained independently associated with clinical decompensation in the final model, whereas MELD

score was excluded. Therapeutic group (timolol or placebo) was unrelated to outcome (Table 2). The results were similar when the analysis was restricted to the subgroup of patients with HCV-related cirrhosis (n = 103), with HVPG, albumin, and BMI being the only variables independently associated with clinical decompensation: HVPG (per 1 mmHg increase HR: 1.19 [95% CI 1.09-1.30], P < 0.0001), albumin (per 1 g/dL decrease HR 2.78 [1.06-7.14], P = 0.04) and BMI (per 1 unit increase HR 1.09 [1.01-1.19], P = 0.03). One hundred eighteen patients (30 normal BMI, 47 overweight, and 41 obese) underwent a second HVPG measurement click here after 1 year of follow-up. The 1-year change in HVPG was linearly correlated with baseline BMI (r = 0.348, P < 0.01) and 12-month BMI (r = 0.306, P < 0.01). Although patients with a normal BMI had a significant reduction in HVPG (mean reduction of 14.3 ± 26.8%; 95% CI 4.3%-23.7%; median reduction 15.2%, P = 0.007 versus baseline), as did overweight patients (mean reduction 7.9 ± 16.4%; 95% CI 2.3%-14.7%; median reduction 11.5%; P = 0.14 versus normal BMI, P = 0.002 versus baseline), obese patients had a slight, nonsignificant increase in HVPG (mean increase of 5.4 ± 32.4%; 95% CI −5.1% to 15.1%); median 0%; P = .004 versus normal BMI; P = 0.

Previously, two Korean studies13,14 reported on the predictors of intraoperative bleeding during gastric ESD. Jang et al. reported that the only factor that correlated with an ‘increased risk’ of bleeding with ESD was the

presence Vemurafenib nmr of gastric malignancy.13 Jeon et al. demonstrated that older age and lesions located in the antrum were associated with a ‘lower frequency’ of bleeding.14 These clinical findings might be associated with vascular factors; the vasculature of malignancies is more tortuous and abundant than that of benign lesions. Moreover, submucosal arteries of the upper third of the stomach are larger than in other areas.8 Therefore, Kuroki et al. revealed this correlation as a model using EUS.12 One of the limitations of ESD is its

technical CP-868596 concentration difficulty. Endoscopists performing ESD need to develop the ability to diagnose margins of the lesion and to perform hemostasis perfectly. Many endoscopists will want to learn how to perform ESD; however, training in an apprentice system is required. Most beginners start ESD at the lower part of stomach, because this part has less vascularity and easier accessibility for the endoscope.15 In the education program, we believe that initial success is important for long-standing success. However, unexpected intraoperative bleeding can cause failure and frustration. Therefore, EUS performed by an expert before the beginner will be helpful to ensure successful ESD. ESD as a curative method for gastric neoplasms should be performed all over the world and is rapidly being introduced. Preventing bleeding is an important factor for successful ESD, and the risk is correlated with the status of submucosal

vascularity. In addition to the role of EUS for diagnosing this website T and N staging of gastric cancer, Kikuchi et al. have shown that we can predict vascular status using EUS.12 We can expect that EUS will also play a useful role as an ESD training tool. In summary, EUS is expecting to improve the feasibility and safety of ESD. “
“Symptomatic gallstone disease (SGSD) induced several inflammatory responses and affected extrahepatic bile ducts. Although the pathology and environmental risk factors of gallstone disease are well documented, immune or inflammatory responses in SGSD development are still inconclusive. Interleukin 18 (IL18) is a pro-inflammatory cytokine that plays an important role in immune, infectious, and inflammatory diseases because of the induction of interferon-γ. In this study, we investigated whether polymorphisms of the IL18 gene were associated with SGSD susceptibility. Genomic DNA was isolated from the whole blood samples of 445 patients with SGSD and 1121 gallstone-free controls. The IL18 rs549908T>G, rs5744247C>G, rs187238G>C, rs1946518T>G, and rs360719A>G polymorphisms were genotyped using predeveloped TaqMan allelic discrimination assay.

h-α-SMA was more strongly expressed than mouse α-SMA, as measured by real-time polymerase chain reaction (PCR), and in drug-treated animals the human isoform of α-SMA but not the murine isoform was down-regulated, suggesting that injected PTFs were still present and functionally active at the end of the experiment, and also that the presence of host/resident myofibroblasts did not significantly affect results (Fig. 6D). In conclusion, we demonstrated that LPA secreted by HCC cells recruits

and activates PTFs, orchestrating their differentiation FK866 to a CAF-like myofibroblastic phenotype which, in turn, accelerates HCC progression. Finally, we aimed to validate these findings in HCC patients. We therefore analyzed LPA serum levels in 60 patients with HCC (30 patients with and 30 without metastases), and in 50 patients with liver cirrhosis. We found that LPA serum levels were higher in HCC compared with liver cirrhosis patients (P < 0.05). Among HCC patients, Dabrafenib LPA serum levels were significantly (P < 0.05) higher in those with metastasis compared with those

without (Fig. 6A). Patients with higher (P < 0.001) serum levels of LPA also have larger HCC tumors (>4 cm) and shorter survival compared with those with lower LPA serum concentrations (Fig. 6B,C). To validate our LPA data in HCC patients, publicly accessible microarray data were analyzed for a correlation between ATX and clinical outcome in HCC patients. ATX expression was significantly increased in HCC patients with more advanced disease, in particular in those with metastatic invasion (P < 0.001) (Fig. 6D),13 and was more strongly expressed in tumoral compared with paired nontumoral tissues (Fig. 6E,F) . In addition, we compared the expression of ATX and LPA receptors in epithelial and stromal components of the same HCC tissues microdissected

using the laser capture microscope technique check details (Fig. 7A,B). We found similar expression levels of ATX in both the epithelial and the stromal component of HCC. However, LPA receptors were essentially expressed in the stromal rather than the epithelial component, indicating the stroma as the main target of the LPA paracrine loop (Fig. 7C). Finally, the ACTA2 gene was significantly expressed in tumoral compared with paired nontumoral tissues (Fig. 7D). This is consistent with publicly accessible microarray data published by Wang (Fig. 7E). Taken together, these data show that the stromal component represents the main target of LPA in patients with HCC, and that α-SMA–positive cells are predominant within the tumor stroma, as shown by the increased expression of the ACTA2 gene.

Interestingly, mitochondria, nuclei, and endoplasmic reticulum remained morphologically unchanged. Cholesterol and neutral lipids (TG and cholesterol esters) were quantified by way of gas/liquid chromatography (Fig. 3). Whereas tetracycline caused no significant changes in TG content after 24 hours,

a six-fold increase was induced by a 50 μM concentration after 14 days. By contrast, BMS-777607 cholesterol and cholesterol esters content remained unchanged. A dose-dependent increase in TG content was also observed, and cholesterol esters were slightly augmented in HepaRG cells treated by amiodarone for 14 days. In addition, phospholipids (phosphatidylethanolamine, phosphatidylcholine, sphingomyelin, phosphatidylserine, and phosphatidylinositol) were measured by way of HPLC in HepaRG cells treated with 20 μM amiodarone for 24 hours or 14 days (Fig. 4). Whereas no significant change was observed in phospholipid content after acute exposure, phosphatidylethanolamine and phosphatidylcholine levels were strongly enhanced, and sphingomyelin, phosphatidylserine, and phosphatidylinositol levels were slightly augmented after 14 days. Impairment of mitochondrial fatty acid oxidation (FAO) is considered one of the major mechanisms of liver steatosis.21 FAO was evaluated by measuring [14C]-labeled acid-soluble

β-oxidation products in HepaRG cells after 24-hour and 14-day treatments using either 20 μM tetracycline or 50 μM amiodarone (Fig. 5). A 20% diminution of FAO was observed after both acute and chronic PLX-4720 amiodarone treatments, and only after chronic tetracycline exposure. To characterize gene expression changes associated with induction of phospholipidosis and steatosis, the transcriptome of HepaRG cells was analyzed after 24-hour and 14-day treatments with 20 μM amiodarone using pangenomic

oligonucleotide microarrays. Significantly modulated genes were extracted with a fold change >1.5 or <−1.5 and P ≤ 0.01 as filters. Their total numbers reached 547 and 594 with up-regulated genes representing 48% and 44%, after 24-hour and 14-day exposure, respectively (Supporting Tables 1 and 2); 176 genes were in common at the two time points. Functional analysis revealed that expression of many genes involved in the regulation of lipid metabolism (including ACOT12, ADFP, ALDH3A1, APOA2, FASN, MOGAT1, SREBP1, find more and THRSP) or related to phospholipidosis (such as LSS, LPIN1, ASML3A, and GDPD3) was significantly altered. Various genes regulating growth/proliferation, cell death, assembly/organization, and inflammation were also substantially deregulated. To validate and complete this microarray analysis, changes in the expression of 29 genes, which are key players in lipid metabolism and/or liver-specific functions, were further examined by way of RT-qPCR in HepaRG cells exposed to several concentrations of amiodarone (5-20 μM), tetracycline (10-100 μM), and oleic acid (100-500 μM) for 24 hours or 14 days. The data are displayed in Table 2.

Interestingly, mitochondria, nuclei, and endoplasmic reticulum remained morphologically unchanged. Cholesterol and neutral lipids (TG and cholesterol esters) were quantified by way of gas/liquid chromatography (Fig. 3). Whereas tetracycline caused no significant changes in TG content after 24 hours,

a six-fold increase was induced by a 50 μM concentration after 14 days. By contrast, GSK-3 activation cholesterol and cholesterol esters content remained unchanged. A dose-dependent increase in TG content was also observed, and cholesterol esters were slightly augmented in HepaRG cells treated by amiodarone for 14 days. In addition, phospholipids (phosphatidylethanolamine, phosphatidylcholine, sphingomyelin, phosphatidylserine, and phosphatidylinositol) were measured by way of HPLC in HepaRG cells treated with 20 μM amiodarone for 24 hours or 14 days (Fig. 4). Whereas no significant change was observed in phospholipid content after acute exposure, phosphatidylethanolamine and phosphatidylcholine levels were strongly enhanced, and sphingomyelin, phosphatidylserine, and phosphatidylinositol levels were slightly augmented after 14 days. Impairment of mitochondrial fatty acid oxidation (FAO) is considered one of the major mechanisms of liver steatosis.21 FAO was evaluated by measuring [14C]-labeled acid-soluble

β-oxidation products in HepaRG cells after 24-hour and 14-day treatments using either 20 μM tetracycline or 50 μM amiodarone (Fig. 5). A 20% diminution of FAO was observed after both acute and chronic PF-6463922 nmr amiodarone treatments, and only after chronic tetracycline exposure. To characterize gene expression changes associated with induction of phospholipidosis and steatosis, the transcriptome of HepaRG cells was analyzed after 24-hour and 14-day treatments with 20 μM amiodarone using pangenomic

oligonucleotide microarrays. Significantly modulated genes were extracted with a fold change >1.5 or <−1.5 and P ≤ 0.01 as filters. Their total numbers reached 547 and 594 with up-regulated genes representing 48% and 44%, after 24-hour and 14-day exposure, respectively (Supporting Tables 1 and 2); 176 genes were in common at the two time points. Functional analysis revealed that expression of many genes involved in the regulation of lipid metabolism (including ACOT12, ADFP, ALDH3A1, APOA2, FASN, MOGAT1, SREBP1, selleck kinase inhibitor and THRSP) or related to phospholipidosis (such as LSS, LPIN1, ASML3A, and GDPD3) was significantly altered. Various genes regulating growth/proliferation, cell death, assembly/organization, and inflammation were also substantially deregulated. To validate and complete this microarray analysis, changes in the expression of 29 genes, which are key players in lipid metabolism and/or liver-specific functions, were further examined by way of RT-qPCR in HepaRG cells exposed to several concentrations of amiodarone (5-20 μM), tetracycline (10-100 μM), and oleic acid (100-500 μM) for 24 hours or 14 days. The data are displayed in Table 2.