Adipose Fatty Acid Binding Protein Research Articles

Development of a humanized anti-FABP4 monoclonal antibody for potential treatment of breast cancer

BackgroundBreast cancer is the most common cancer in women diagnosed in the U.S. and worldwide. Obesity increases breast cancer risk without clear underlying molecular mechanisms. Our studies demonstrate that circulating adipose fatty acid binding protein (A-FABP, or FABP4) links obesity-induced dysregulated lipid metabolism and breast cancer risk, thus potentially offering a new target for breast cancer treatment.MethodsWe immunized FABP4 knockout mice with recombinant human FABP4 and screened hybridoma clones with specific binding to FABP4. The potential effects of antibodies on breast cancer cells in vitro were evaluated using migration, invasion, and limiting dilution assays. Tumor progression in vivo was evaluated in various types of tumorigenesis models including C57BL/6 mice, Balb/c mice, and SCID mice. The phenotype and function of immune cells in tumor microenvironment were characterized with multi-color flow cytometry. Tumor stemness was detected by ALDH assays. To characterize antigen-antibody binding capacity, we determined the dissociation constant of selected anti-FABP4 antibodies via surface plasmon resonance. Further analyses in tumor tissue were performed using 10X Genomics Visium spatial single cell technology.ResultsHerein, we report the generation of humanized monoclonal antibodies blocking FABP4 activity for breast cancer treatment in mouse models. One clone, named 12G2, which significantly reduced circulating levels of FABP4 and inhibited mammary tumor growth, was selected for further characterization. After confirming the therapeutic efficacy of the chimeric 12G2 monoclonal antibody consisting of mouse variable regions and human IgG1 constant regions, 16 humanized 12G2 monoclonal antibody variants were generated by grafting its complementary determining regions to selected human germline sequences. Humanized V9 monoclonal antibody showed consistent results in inhibiting mammary tumor growth and metastasis by affecting tumor cell mitochondrial metabolism.ConclusionsOur current evidence suggests that targeting FABP4 with humanized monoclonal antibodies may represent a novel strategy for the treatment of breast cancer and possibly other obesity- associated diseases.

Association study between SNP markers located in meat quality candidate genes with intramuscular fat content in an endangered dual-purpose cattle population.

The aim of this study was to associate single nucleotide polymorphisms (SNP) of the bovine calcium-activated neutral protease µ-calpain, calpastatin, diacylglycerol-O-acyltransferase, adipose fatty acid binding protein, retinoic acid receptor-related orphan receptor C (RORC), and thyroglobulin (TG) gene with intramuscular fat content (IMF). Therefore, 542 animals of the cattle breed "Rotes Höhenvieh" (RHV) were phenotyped for IMF. Genotyping of the animals was performed using polymerase chain reaction-restriction fragment length polymorphism tests for six SNP from candidate genes for meat quality traits. In addition, we calculated allele substitution and dominance effects on IMF. A subgroup of animals (n = 44, reduced dataset) with extraordinary high IMF was analyzed separately. The mean IMF content was 2.5% (SD: 2.8) but ranged from 0.02% to 23.9%, underlining the breeds' potential for quality meat production. Allele and genotype frequencies for all SNP were similar in the complete and reduced dataset. Association analyses in the complete dataset revealed the strongest effects of RORC on IMF (P = 0.075). The log-transformed least-squares mean for IMF of genotype g.3290GG was 0.45 ± 0.16, 0.26 ± 0.14 for genotype g.3290GT, and 0.32 ± 0.14 for genotype g.3290TT. In the reduced dataset, we found a significant effect (P < 0.05) of the g.422C>T-SNP of TG on IMF, with highest IMF for genotype CT (0.91 ± 0.17), lowest IMF for genotype TT (0.37 ± 0.25), and medium IMF for genotype CC (0.59 ± 0.16; log-transformed values). Compared to the complete dataset, allele substitution effects increased in the reduced dataset for most of the SNP, possibly due to the selective genotyping strategy, with focus on animals with highest IMF implying strong phenotypic IMF contrast. Dominance effects were small in both datasets, related to the high heritability of IMF. Results indicated RHV breed particularities regarding the effects of meat quality genes on IMF. An explanation might be the breeding history of RHV with focus on adaptation and resilience in harsh outdoor systems. Consequently, it is imperative to develop breed-specific selection strategies. Allele substitution and dominance effects were in a similar direction in both datasets, suggesting the same breeding approaches for different RHV strains in different regions. Nevertheless, a selective genotyping approach (reduced dataset), contributed to more pronounced genotype effect differences on IMF and dominance values.

FABP4: A New Player in Obesity-Associated Breast Cancer

Obesity is known to increase breast cancer incidence and mortality, but the underlying mechanisms remain unsolved. Recent studies demonstrate that adipose fatty acid binding protein (FABP4) promotes obesity-associated breast cancer development, thus suggesting FABP4 as a novel player linking obesity and breast cancer risk.

Circulating Adipose Fatty Acid Binding Protein Is a New Link Underlying Obesity-Associated Breast/Mammary Tumor Development

It is clear that obesity increases the risk of many types of cancer, including breast cancer. However, the underlying molecular mechanisms by which obesity is linked to cancer risk remain to be defined. Herein, we report that circulating adipose fatty acid binding protein (A-FABP) promotes obesity-associated breast cancer development. Using clinical samples, we demonstrated that circulating A-FABP levels were significantly increased in obese patients with breast cancer in comparison with those without breast cancer. Circulating A-FABP released by adipose tissue directly targeted mammary tumor cells, enhancing tumor stemness and aggressiveness through activation of the IL-6/STAT3/ALDH1 pathway. Importantly, genetic deletion of A-FABP successfully reduced tumor ALHD1 activation and obesity-associated mammary tumor growth and development in different mouse models. Collectively, these data suggest circulating A-FABP as a new link between obesity and breast cancer risk, thereby revealing A-FABP as a potential new therapeutic target for treatment of obesity-associated cancers.

Cardiometabolic Biomarkers and Their Temporal Patterns Predict Poor Outcome in Chronic Heart Failure (Bio-SHiFT Study).

Multiple hormonal and metabolic alterations occur in chronic heart failure (CHF), but their proper monitoring during clinically silent progression of CHF remains challenging. Hence, our objective was to explore whether temporal patterns of six emerging cardiometabolic biomarkers predict future adverse clinical events in stable patients with CHF. In 263 patients with CHF, we determined the risk of a composite end point of heart failure hospitalization, cardiac death, left ventricular assist device implantation, and heart transplantation in relation to serially assessed blood biomarker levels and slopes (i.e., rate of biomarker change per year). During 2.2 years of follow-up, we repeatedly measured IGF binding proteins 1, 2, and 7 (IGFBP-1, IGFBP-2, IGFBP-7), adipose fatty acid binding protein 4 (FABP-4), resistin, and chemerin (567 samples in total). Serially measured IGFBP-1, IGFBP-2, IGFBP-7, and FABP-4 levels predicted the end point [univariable hazard ratio (95% CI) per 1-SD increase: 3.34 (2.43 to 4.87), 2.86 (2.10 to 3.92), 2.45 (1.91 to 3.13), and 2.46 (1.88 to 3.24), respectively]. Independently of the biomarkers' levels, their slopes were also strong clinical predictors [per 0.1-SD increase: 1.20 (1.11 to 1.31), 1.27 (1.14 to 1.45), 1.23 (1.11 to 1.37), and 1.27 (1.12 to 1.48)]. All associations persisted after multivariable adjustment for patient baseline characteristics, baseline N-terminal pro-hormone brain natriuretic peptide and cardiac troponin T, and pharmacological treatment during follow-up. The temporal patterns of IGFBP-1, IGFBP-2, IGFBP-7, and adipose FABP-4 predict adverse clinical outcomes during outpatient follow-up of patients with CHF and may be clinically relevant as they could help detect more aggressive CHF forms and assess patient prognosis, as well as ultimately aid in designing more effective biomarker-guided therapy.

Circulating adipose fatty acid binding protein is a new link underlying obesity-associated breast/ mammary tumor development

Circulating adipose fatty acid binding protein is a new link underlying obesity-associated breast/ mammary tumor development

Saturated Fatty Acids Induce Ceramide-associated Macrophage Cell Death

Macrophages highly express epidermal fatty acid-binding protein and adipose fatty acid-binding protein. They actively uptake saturated and unsaturated fatty acids, which might play a critical role in regulating their immune functions. Numerous studies have shown that various fatty acids, saturated or unsaturated, may possess different impacts on cell growth and function. However, the approaches used for fatty acid preparation vary, which may lead to non-physiological results. Serum albumin, a natural carrier for fatty acids in mammalian peripheral blood, is recommended for forming a conjugate complex with the sodium salt of fatty acids to study fatty acid function in mammalian cells, thus minimizing the toxicity of fatty acid soap. Thus, a simple, relatively quick heating and sonicating method is developed and presented here for BSA-fatty acid conjugate formation. We describe a protocol using saturated fatty acids, especially stearic acids to induce severe cell death in mouse bone-marrow derived macrophages. We further demonstrate that saturated fatty acid-induced cell death is positively associated with accumulated cellular ceramide levels. This method can be extended for studies of the impact of fatty acid on other mammalian cells.

Adipose Fatty Acid Binding Protein Promotes Saturated Fatty Acid-Induced Macrophage Cell Death through Enhancing Ceramide Production.

Macrophages play a critical role in obesity-associated chronic inflammation and disorders. However, the molecular mechanisms underlying the response of macrophages to elevated fatty acids (FAs) and their contribution to metabolic inflammation in obesity remain to be fully elucidated. In this article, we report a new mechanism by which dietary FAs, in particular, saturated FAs (sFAs), are able to directly trigger macrophage cell death. We demonstrated that excess sFAs, but not unsaturated FAs, induced the production of cytotoxic ceramides (Cers) in macrophage cell lines. Most importantly, expression of adipose FA binding protein (A-FABP) in macrophages facilitated metabolism of excess sFAs for Cer synthesis. Inhibition or deficiency of A-FABP in macrophage cell lines decreased sFA-induced Cer production, thereby resulting in reduced cell death. Furthermore, we validated the role of A-FABP in promoting sFA-induced macrophage cell death with primary bone marrow-derived macrophages and high-fat diet-induced obese mice. Altogether, our data reveal that excess dietary sFAs may serve as direct triggers in induction of Cer production and macrophage cell death through elevated expression of A-FABP, thus establishing A-FABP as a new molecular sensor in triggering macrophage-associated sterile inflammation in obesity.

Adipose Fatty Acid Binding Protein Promotes Saturated Fatty Acid-induced Cell Death through Ceramide Production in Macrophages

Abstract Macrophages play a critical role in obesity-associated chronic inflammation and disorders. However, the molecular mechanisms of how macrophages respond to elevated fatty acids (FAs) and contribute to sterile inflammation in obesity remain to be fully elucidated. Here, we report a new mechanism by which dietary FAs, in particular saturated FAs, were able to directly trigger macrophage cell death. We demonstrated that excess saturated FAs, but not unsaturated FAs, induced the production of cytotoxic ceramides in macrophages. Most importantly, expression of adipose fatty acid binding protein (A-FABP) in macrophages facilitated metabolism of excess saturated FAs for ceramide synthesis. Inhibition or deficiency of A-FABP in macrophages decreased saturated FA-induced ceramide production, thereby resulting in reduced cell death. Furthermore, A-FABP-deficient obese mice exhibited increased percentage of CD36+ macrophages in peripheral as compared to WT control mice. Altogether, our data suggest that excess saturated FAs in diets serve as new triggers in inducing ceramide production and macrophage cell death through elevated expression of A-FABP, thus contributing to sterile inflammation in obesity.

Generation of porcine fibroblasts overexpressing 11β-HSD1 with adipose tissue-specific aP2 promoter as a porcine model of metabolic syndrome

Metabolic syndrome arises from a combination of disorders that increase the risk of cardiovascular disease and diabetes. In previous studies, it was observed that overexpression of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) induced obesity and the insulin resistance that accompanies metabolic syndrome in rodent adipose tissue. Based on these observations, it was hypothesized that overexpression of 11β-HSD1 may be suitable for the generation of a porcine model of metabolic syndrome. It was evaluated that promoter activities of the porcine adipose fatty acid-binding protein (aP2) gene generates adipose tissue-specific 11β-HSD1 expression. In adipose tissue, the maximum promoter activity (-2,826 to +51 nt) of aP2 was 200-fold higher than that of a promoterless construct. In addition, 11β-HSD1 transcriptional levels were significantly increased following the introduction of the aP2 promoter into 3T3‑L1 adipocytes. These observations indicate that the aP2 promoter may facilitate 11β-HSD1 overexpression in porcine adipose tissue. Transgenic fibroblasts were generated containing 11β-HSD1 cDNA controlled by the aP2 promoter with two screening markers, green fluorescence protein and a neomycin-resistance gene. It was hypothesized that transgenic fibroblasts may be useful for generating a porcine model of metabolic syndrome.

Serum adipocyte fatty acid-binding protein in the critically ill

Sepsis due to unabated inflammation is common. Increased production of pro-inflammatory cytokines, free radicals, and eicosanoids has been detected in sepsis and other critical illnesses but could also be due to decreased synthesis and release of anti-inflammatory molecules. Increased serum adipose-fatty acid-binding protein (A-FABP) levels can cause insulin resistance and have been reported in the critically ill, serve as a marker of prognosis, and thus link metabolic homeostasis and inflammation. A-FABP can be linked to the expression of Toll-like receptors, macrophage activation, synthesis and release of pro-inflammatory cytokines interleukin-6 and tumor necrosis factor-alpha, activation of cyclooxygenase 2 (COX-2) expression, and eicosanoid synthesis, events that can cause insulin resistance and initiation and progression of inflammation and sepsis. Unsaturated fatty acids and their anti-inflammatory products, such as lipoxins, resolvins, and protectins, may suppress A-FABP expression, inhibit macrophage and COX-2 activation, and decrease production of pro-inflammatory cytokines and ultimately could lead to a decrease in insulin resistance and resolution of inflammation and recovery from sepsis. Serial measurement of these pro- and anti-inflammatory molecules and correlation of their levels to the progression to or recovery from (or both) sepsis and other inflammatory processes may form a new approach to predict prognosis in inflammatory conditions and eventually could lead to the development of new therapeutic strategies.

Abstract 190: Activation of Angiotensin Type 2 Receptor Alters High-fat Diet-Induced Lipid Metabolism and Rescues Adipocyte Size Increase in Young Male Mice

Recent studies demonstrated that the genetic deletion of the angiotensin type 2 receptor (AT2R) decreases adipocyte size and controls obesity, a major risk factor for metabolic syndrome. Current study was designed to investigate the effect of pharmacological activation of AT2R on adipocyte size. Male C57BL/6 mice (4-weeks old) were pretreated with the AT2R agonist C21 (0.3 mg/kg, daily i.p.) for 4 days. Thereafter the animals were placed on normal chow diet (ND) or high-fat diet (HFD) with concurrent drug treatment for next 10 days. In 10 days, the HFD significantly increased the body weight (ND:22+0.26 g vs HFD:24+0.40 g), epididymal fat pads weight (ND:0.23±0.03 g vs HFD:0.42±0.03 g), epididymal adipocyte size (ND:1407+176 μm2 vs HFD:2358+96 μm2), plasma free fatty acid (FFA) (ND:0.63+0.04 nmol/μL vs HFD:0.84+0.06 nmol/μL) and triglyceride (TG) (ND:0.31+0.01 nmol/μL vs HFD:0.40+0.03 nmol/μL). The pharmacological activation of AT2R rescued the HFD-induced increase in body weight (22+0.39 g), epididymal fat pads weight (0.30±0.02 g), epididymal adipocyte size (1831+95 μm2), plasma FFA (0.56+0.03 nmol/μL) and TG (0.18+0.02 nmol/μL). On one hand, HFD increased the protein expressions (measured by western blot) of the epididymal white adipose tissue (WAT) lipogenic regulators namely lipoprotein lipase (LPL) (142%), adipose fatty acid binding protein (FABP4) (199%), fatty acid synthase (FASN) (225%), peroxisome proliferator-activated receptor gamma (PPAR-γ) (176%). On the other hand, a differential regulation in the protein expressions of the epididymal WAT lipolytic regulators was observed. The HFD caused decrease in the protein expression of epididymal WAT adipose triglyceride lipase (ATGL) (62%) while increase in hormone-sensitive lipase (HSL) (191%) protein expression. C21 treatment altered all the HFD-induced changes in the protein expressions of LPL (83%), FABP4 (79%), FASN (54%), PPAR-γ (49%), ATGL (167%) and HSL (50%). In conclusion, the pharmacological activation of AT2R alters the HFD-induced lipid metabolism (lipogenesis and lipolysis) and rescues adipocyte size increase. This is the first study suggesting that the pharmacological activation of AT2R may serve as therapeutic strategy for controlling adipocyte size and obesity.

The calcium-sensing receptor promotes adipocyte differentiation and adipogenesis through PPARγ pathway

Adipocyte differentiation and adipogenesis are closely related to obesity and obesity-induced metabolic disorders. The calcium-sensing receptor (CaSR) has been reported to play an antilipolytic role in human adipocyte and regulate cell differentiation in many tissues. However, the effects of CaSR on adipocyte differentiation and adipogenesis have not been clarified. In the study, we observed that activation of CaSR significantly promoted adipocyte differentiation and adipogenesis in human SW872 adipocytes. Gene expression analysis revealed that the CaSR activation increased the transcription factor proliferator-activated receptor γ (PPARγ) and its downstream genes including CCAAT element binding protein α (C/EBPα), adipose fatty acid-binding protein (aP2), and lipoprotein lipase. The activity of glycerol-3-phosphate dehydrogenase was also increased after the stimulation of CaSR. In addition, levels of cyclic AMP and calcium which have been shown to regulate PPARγ gene expression were significantly affected by the activation of CaSR. These effects could be suppressed by CaSR small interfering RNA (CaSR-siRNA). In conclusion, our findings suggest that activation of CaSR promotes differentiation and adipogenesis in adipocytes, which might be achieved by upregulating PPARγ and its downstream gene expressions. Therefore, CaSR in adipocytes may be involved in the pathogenesis of obesity by promoting adipocyte differentiation and adipogenesis.

Unsaturated Fatty Acids Repress the Expression of Adipose Fatty Acid Binding Protein (aP2) in RAW 264.7 Macrophages

Adipocyte fatty acid binding protein, aP2, present in macrophages has been implicated in the integration of lipid metabolism and inflammatory response, contributing to development of insulin resistance and atherosclerosis. We investigated the modulation of aP2 expression by inflammatory insults and fatty acids in RAW 264.7 macrophages. When the cells were incubated with lipopolysaccharides (LPS; 100 ng/ml) or 10 ng/ml of tumor necrosis factor α for 18 h, aP2 mRNA and protein levels were drastically increased. Unsaturated fatty acids (100 μM of 18:1, 18:2, 18:3, 20:5 in complex with BSA), but not saturated fatty acids (16:0), significantly repressed the basal aP2 expression and abolished induction of aP2 expression by LPS. Trichstatin A (TSA), a histone deacetylase (HDAC) inhibitor, increased aP2 mRNA levels but abolish the repressive effect of 18:2 on aP2 expression in unstimulated and LPS‐stimulated macrophages. Depletion of HDAC3 or c‐Jun by siRNA significantly increased aP2 expression in unstimulated macrophages. In summary, our data suggest that basal aP2 expression in macrophages is likely to be repressed by c‐Jun in association with a corepressor complex containing HDAC3 and unsaturated fatty acids may inhibit the basal as well as LPS‐induced aP2 expression by modulating the recruitment or activity of co‐repressor complex to the promoter of aP2.Grant Funding Source: Faculty Seed Grant from University of Nebraska‐Lincoln Research Council

Association analysis of single nucleotide polymorphisms in DGAT1, TG and FABP4 genes and intramuscular fat in crossbred Bos taurus cattle

Previous studies have indicated that single nucleotide polymorphisms (SNPs) in the diacylglycerol-O-acyltransferase1 (DGAT1), thyroglobulin (TG) and adipose fatty acid binding protein (FABP4) genes are associated with intramuscular fat (IMF) levels or marbling scores in beef. The objectives were to estimate the frequency of SNPs in these candidate genes in purebred Irish cattle (n=459) and to determine if individual SNPs are associated with IMF values of longissimus thoracis et lumborum (LTL) and semimembranosus (SM) muscle of crossbred animals (n=138). Results indicated no significant association between the SNPs and IMF, despite the power of this study being sufficient to detect an association with SNPs in the DGAT1 and FABP4 genes. The results confirm the lack of an association found by many other studies and suggest that these SNPs are not influential on the divergent IMF levels in the crossbred population tested.

Association of adipocyte fatty acid-binding protein with obesity and metabolism syndrome

Adipocyte fatty acid-binding protein(A-FABP)belongs to the fatty acid-binding protein super-family, and it is highly expressed in adipose tissue.The physiologic function of A-FABP is involved in the intracellular trafficking and targeting of fatty acids inside cells.Recent research indicated that this protein may be all important regulator of systemic insulin sensitivity as well as lipid and glucose metabolism,indicating that A-FABP participates in the pathogenesis of obesity and metabolic syndrome. Key words: Adipose fatty acid binding protein; Obesity; Metabolic syndrome

Osthole, a potential antidiabetic agent, alleviates hyperglycemia in db/db mice

Osthole is an agent isolated from Cnidium monnieri (L.) Cusson and Angelica pubescens and has been used to treat several diseases, including metabolic syndromes. To investigate the hypoglycemic effects of osthole in diabetic db/db mice and the underlying mechanisms of these effects by in vitro assay, diabetic db/db mice and cell experiments were utilized to understand its possible effects. Osthole significantly activated both PPARα and PPARγ in a dose-dependent manner based on the results of the transition transfection assay. The activation of PPARα and PPARγ by osthole also resulted in an increase in the expression of PPAR target genes such as PPAR itself, adipose fatty acid-binding protein 2, acyl-CoA synthetases, and carnitine palmitoyltransferase-1A. In vitro results suggested that osthole might be a dual PPARα/γ activator, but its chemical structure differed from that of the thiazolidinedione class of antidiabetic drugs. In addition, osthole markedly activated the AMP-activated protein kinase and its downstream acetyl CoA carboxylase molecules by increasing their phosphorylation levels. Finally, obese diabetic db/db mice were treated with osthole by different administered routes, and osthole was found to markedly reduce blood glucose level. Interestingly, osthole did not reduce the blood insulin or lipid levels, two phenomena that did occur in animals treated with insulin sensitizers like PPAR agonists. These results suggest that osthole can alleviate hyperglycemia and could be potentially developed into a novel drug for treatment of diabetes mellitus.

Identification of a Functional Polymorphism at the Adipose Fatty Acid Binding Protein Gene (FABP4) and Demonstration of its Association with Cardiovascular Disease: A Path to Follow

Fatty acid binding proteins (FABPs) are proteins that reversibly bind fatty acids and other lipids. So far, nine tissue-specific cytoplasmic FABPs have been identified. Adipose tissue FABP (FABP4) has been suggested to be a bridge between inflammation and other pathways related to the metabolic syndrome. In this regard, genetic variability at the FABP4 locus has been shown to be associated with plasma lipid levels, type 2 diabetes, and coronary heart disease risk.

Effect of dermcidin, an antimicrobial peptide, on body fat mobilization in normal mice

Dermcidin (DCD), an antimicrobial peptide that is secreted by sweat glands, is reportedly a human homolog of mouse proteolysis-inducing factor. This study was conducted to investigate the effect of DCD on body fat mobilization. The expression level of DCD in the livers of Ad-DCD-injected mice was higher than in those of Ad-beta-galactosidase (Ad-beta-gal)-injected mice 7 days after injection. In addition, injection with the Ad-DCD virus led to decreased body weight and epididymal fat mass when compared with controls. The plasma triglyceride level was decreased, whereas the free fatty acid and glycerol levels were increased in the Ad-DCD-injected group. Epididymal adipose tissues obtained from Ad-DCD-injected mice consisted of smaller adipocytes than tissues obtained from Ad-beta-gal-injected mice. The gene expression profiles revealed an upregulation of hormone-sensitive lipase and adipose fatty acid-binding protein, both of which are involved in adipocyte lipolysis, in Ad-DCD-injected mice, and this lipolytic effect of DCD paralleled the increase of circulating tumor necrosis factor-alpha (TNF-alpha) level that was observed. The perilipin levels in adipose tissue were decreased in Ad-DCD-injected mice when compared with those of the control mice. Taken together, these results suggest that DCD-mediated body fat reduction might occur as a result of TNF-alpha-induced downregulation of perilipin in adipose tissue.

The establishment of a novel non-alcoholic steatohepatitis model accompanied with obesity and insulin resistance in mice

Non-alcoholic steatohepatitis (NASH) is a hepatic manifestation of the metabolic syndrome that can progress to liver cirrhosis. The major aim of this study was to establish a novel NASH mouse model accompanied by obesity and insulin resistance, then explore the molecular mechanisms of NASH and evaluate the effects of both the peroxisome proliferator-activated receptor α (PPARα) agonist fenofibrate and the PPARγ agonist rosiglitazone in this established NASH model. The novel model was induced in C57BL/6 mice by 23 weeks of ad libitum feeding of a modified high-fat diet (mHFD), with lower methinione and choline and higher fat content. In comparison to the controls, the model animals developed pronounced obesity, dyslipidemia and insulin resistance. Marked liver lesions characterized by severe steatosis, inflammation, fibrosis, increased hepatic triglyceride content, and elevated serum alanine aminotransferase (ALT) levels were observed in the models. In this novel model, treatment with fenofibrate or rosiglitazone significantly improved insulin sensitivity and corrected dyslipidemia; however, fenofibrate was more effective than rosiglitazone in improving hepatic morphology and ALT levels. Further study showed that long-term feeding of mHFD significantly increased expression of mRNA for hepatic PPARγ, adipose fatty acid binding protein (ap2) and CD36 and suppressed expression of mRNA for hepatic PPARα and carnitine palmitoyl transferase-1a (CPT-1a). These results showed the successful establishment of the combined NASH and obese-insulin resistance mouse model. Additionally, aberrant expressions of hepatic PPARα and PPARγ may play a major role in the pathogenesis of NASH by affecting hepatic lipogenesis and fatty acid oxidation in this novel model.