Hepatic Fatty Acid Oxidation Research Articles

Farnesol Decreases Serum Triglycerides in Rats: Identification of Mechanisms Including Up‐Regulation of PPARα and Down‐Regulation of Fatty Acid Synthase in Hepatocytes

Obesity is associated with impaired fatty acid (FA) oxidation and increased de novo hepatic lipogenesis that may contribute to the development of hypertriglyceridemia, an important risk factor for the development of cardiovascular disease. Strategies to improve hepatocyte FA metabolism, including dietary interventions, are therefore important for the prevention of obesity-associated co-morbidities. Farnesol is consumed in the diet as a component of plant products. In the present study, we administered farnesol orally to rats for seven days and found significantly reduced serum triglyceride concentrations compared with controls. Potential mechanisms underlying the hypotriglyceridemic effect of farnesol were investigated using clone-9 cultured rat hepatocytes. Farnesol significantly upregulated expression of peroxisome proliferator-activated receptor alpha (PPARalpha) and the PPARalpha-regulated genes fatty acyl-CoA oxidase and carnitine palmitoyl transferase 1a, suggesting that increased hepatic FA oxidation may contribute to serum triglyceride lowering in rats. Farnesol did not change SREBP-1c mRNA levels, but significantly down-regulated fatty acid synthase (FAS) mRNA and protein levels and activity, indicating that attenuated lipogenesis may also contribute to hypotriglyceridemic effects of farnesol in vivo. Rescue experiments revealed that down-regulation of FAS by farnesol was not related to activation of PPARalpha, but rather was caused by a 9-cis retinoic acid mediated mechanism that involved down-regulation of retinoid X receptor beta. Diets rich in plant products are associated with a lower risk of cardiovascular disease. Our findings suggest that farnesol may contribute to this protective effect by lowering serum TG levels.

Apolipoprotein AII Is a Regulator of Very Low Density Lipoprotein Metabolism and Insulin Resistance

Apolipoprotein AII (apoAII) transgenic (apoAIItg) mice exhibit several traits associated with the insulin resistance (IR) syndrome, including IR, obesity, and a marked hypertriglyceridemia. Because treatment of the apoAIItg mice with rosiglitazone ameliorated the IR and hypertriglyceridemia, we hypothesized that the hypertriglyceridemia was due largely to overproduction of very low density lipoprotein (VLDL) by the liver, a normal response to chronically elevated insulin and glucose. We now report in vivo and in vitro studies that indicate that hepatic fatty acid oxidation was reduced and lipogenesis increased, resulting in a 25% increase in triglyceride secretion in the apoAIItg mice. In addition, we observed that hydrolysis of triglycerides from both chylomicrons and VLDL was significantly reduced in the apoAIItg mice, further contributing to the hypertriglyceridemia. This is a direct, acute effect, because when mouse apoAII was injected into mice, plasma triglyceride concentrations were significantly increased within 4 h. VLDL from both control and apoAIItg mice contained significant amounts of apoAII, with approximately 4 times more apoAII on apoAIItg VLDL. ApoAII was shown to transfer spontaneously from high density lipoprotein (HDL) to VLDL in vitro, resulting in VLDL that was a poorer substrate for hydrolysis by lipoprotein lipase. These results indicate that one function of apoAII is to regulate the metabolism of triglyceride-rich lipoproteins, with HDL serving as a plasma reservoir of apoAII that is transferred to the triglyceride-rich lipoproteins in much the same way as VLDL and chylomicrons acquire most of their apoCs from HDL.

S1894 Physical Activity and Short-Term Inactivity Modulate Hepatic Fatty Acid Oxidation and Lipogenesis Proteins in Otsuka Long-Evans Tokushima Fatty (Oletf) Rats

S1894 Physical Activity and Short-Term Inactivity Modulate Hepatic Fatty Acid Oxidation and Lipogenesis Proteins in Otsuka Long-Evans Tokushima Fatty (Oletf) Rats

Effects of soy protein and isoflavone on hepatic fatty acid synthesis and oxidation and mRNA expression of uncoupling proteins and peroxisome proliferator-activated receptor γ in adipose tissues of rats

Soy protein rich in isoflavones profoundly affects lipid metabolism in experimental animals. To distinguish the roles of the protein and isoflavone components of a soy protein preparation in regulating lipid metabolism, we compared the effects of diets containing methanol-washed soy protein low in isoflavone supplemented with a 0-, 0.5- and 4-g/kg isoflavone preparation on hepatic fatty acid metabolism and adipose tissue gene expression in rats. Diets containing soy protein irrespective of the isoflavone levels decreased the activities and mRNA expression of enzymes involved in hepatic fatty acid synthesis to similar levels. Methanol-washed soy protein compared to casein increased the mRNA expression of peroxisome proliferator-activated receptor (PPAR) α, and supplementing the soy protein diet with isoflavone further increased this parameter dose-dependently. However, methanol-washed soy protein compared to casein was totally ineffective in altering the activities and mRNA levels of enzymes involved in fatty acid oxidation. Supplementation of soy protein diets with isoflavone slightly increased these parameters. The mRNA level of uncoupling protein (UCP) 1 in brown adipose tissue was significantly increased and mRNA levels of UCP2 and 3, and PPARγ2 tended to be higher in rats fed methanol-washed soy protein not supplemented with isoflavone than in the animals fed casein. Adding isoflavone to the soy protein diets dose-dependently increased these parameters. These results suggested that the protein rather than isoflavone component is primarily responsible for the physiological activity of soy protein rich in isoflavones in reducing hepatic lipogenesis. However, isoflavones may have a role in regulating heptic fatty acid oxidation and adipose tissue gene expression.

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.

Transplacental induction of fatty acid oxidation in newborn pigs by clofibrate

This study examined the effect of maternal feeding of clofibrate on fatty acid oxidation in newborn pigs. Pregnant sows were fed a commercial diet with or without 0.8% clofibrate for 7 days. Blood samples were collected from the utero‐ovarian artery of the sows and the umbilical vein of the intact term fetuses as they were removed from the sows by c‐section on day 113 of gestation. HPLC analysis clearly identified clofibric acid was present in the plasma of the clofibrate fed sow and the fetuses. The maternal‐fed clofibric acid had no impact on the liver weight of the newborn pigs, but hepatic fatty acid oxidation examined in fresh homogenates showed that clofibrate significantly increased 14C‐accumulation in CO2 and ASP by 2.9‐fold from [1‐14C]‐oleic acid and 1.6‐fold from [1‐14C]‐lignoceric acid. Correspondingly, hepatic carnitine palmitoyltransferase (CPT) and fatty acid oxidase (FAO) activities were increased also by 36% and 42% compared to the newborn pigs from the control sow. These data demonstrate that clofibrate crosses the placental membrane of the sow and enters fetal circulation to induce hepatic fatty acid oxidation by increasing CPT and FAO activities of the newborn. Supported by CSREES, USDA NRI program award 2007‐35206‐17897.

Fast/Glycolytic Muscle Fiber Growth Reduces Fat Mass and Improves Metabolic Parameters in Obese Mice

In contrast to the well-established role of oxidative muscle fibers in regulating whole-body metabolism, little is known about the function of fast/glycolytic muscle fibers in these processes. Here, we generated a skeletal muscle-specific, conditional transgenic mouse expressing a constitutively active form of Akt1. Transgene activation led to muscle hypertrophy due to the growth of type IIb muscle fibers, which was accompanied by an increase in strength. Akt1 transgene induction in diet-induced obese mice led to reductions in body weight and fat mass, resolution of hepatic steatosis, and improved metabolic parameters. Akt1-mediated skeletal muscle growth opposed the effects of a high-fat/high-sucrose diet on transcript expression patterns in the liver and increased hepatic fatty acid oxidation and ketone body production. Our findings indicate that an increase in fast/glycolytic muscle mass can result in the regression of obesity and metabolic improvement through its ability to alter fatty acid oxidation in remote tissues.

Dissociation of mercaptoacetate’s effects on feeding and fat metabolism by dietary medium- and long-chain triacylglycerols in rats

Objective and Methods Mercaptoacetate (MA) inhibits hepatic fatty acid oxidation (FAO) and stimulates feeding in rats fed fat-rich diets. To test whether the feeding stimulation by MA depends on hepatic FAO, we compared the effects of intraperitoneally injected MA (45.6 mg/kg body weight) with saline in rats fed diets containing 18% predominately long-chain triacylglycerols (LCTs; ≥90% 16 C) or 18% medium-chain triacylglycerols (MCTs; 51% 10–12 C). We hypothesized that, because medium-chain fatty acids reach the liver and are oxidized faster than long-chain fatty acids, if MA’s feeding-stimulatory effect depends on hepatic FAO, MA should stimulate feeding more in MCT-fed rats than in LCT-fed rats. Results Although MA injected in mid-light phase stimulated feeding similarly in MCT- and LCT-fed rats, MA injected at light onset initially stimulated food intake (1 h) only in LCT- and not in MCT-fed rats. To investigate MA’s metabolic effects during the initial hour, rats were sacrificed 30 min after light-onset injections. At this time plasma β-hydroxybutyrate appeared to be higher in MCT- than in LCT-fed rats and to be increased by MA. In a final experiment, MA did not affect fatty acid content in liver and duodenum tissues but increased fatty acids in duodenal tissue mitochondria from 12 h-fasted rats fed chow. Conclusion In light-onset tests, adaptation to the MCT diet increased hepatic FAO but not the feeding-stimulatory effect of MA in comparison with adaptation to the LCT diet, suggesting that at this time MA does not act in the liver to stimulate feeding or that this effect is not due to FAO inhibition. Inhibition of duodenal mitochondrial FAO may be another metabolic process through which MA stimulates feeding.

Liver and muscle metabolic changes induced by dietary energy content and genetic selection in rainbow trout (Oncorhynchus mykiss)

We combined genetic selection and dietary treatment to produce a model to study metabolic pathways involved in genetic and nutritional control of fat deposition in fish muscle. Two experimental lines of rainbow trout, selected for a lean (L) or fat (F) muscle, were fed with diets containing either 10 or 23% lipids from the first feeding, up to 6 mo. At the end of the feeding trial, trout were distinguished by very different muscle fat content (from 4.2 to 10% wet weight), and line x diet interactions were observed for parameters related to fat storage. We analyzed the activity and gene expression of key enzymes involved in lipid metabolism (fatty acid synthase, hydroxyacyl-CoA dehydrogenase, carnitine palmitoyltransferase 1 isoforms, and peroxisome proliferator-activated receptor alpha) and glycolysis (hexokinase 1 and pyruvate kinase) as well as energy production (isocitrate dehydrogenase, citrate synthase, and cytochrome oxidase) in the liver and the white muscle of rainbow trout. The lipid-rich diet repressed the activity of the lipogenic enzymes and stimulated enzymes involved in fatty acid oxidation and glycolysis in liver but had little effect on muscle enzymes assessed in this study. Regarding the selection effect, enzyme activity and expression suggest that compared with the L line, the F line presented reduced hepatic fatty acid oxidation as well as reduced mitochondrial oxidative capacities and enhanced glucose utilization in both liver and muscle. Very few line x diet interactions were found, suggesting that the two factors (i.e., dietary energy content and selection) used in this study to modify muscle lipid content exerted some additive but mostly independent effects on these metabolic actors.

Utilization of different dietary lipid sources at high level in herbivorous grass carp (Ctenopharyngodon idella): mechanism related to hepatic fatty acid oxidation

Herbivorous grass carp (Ctenopharyngodon idella) has been reported to exhibit low capacity to utilize high dietary lipid, but different lipid sources might affect this limited capacity. In order to compare the effects of different lipid sources with different lipid levels, juvenile grass carp were fed one of nine diets containing three oils [lard, plant oil mixed by maize and linseed oil, and n-3 high unsaturated fatty acid-enriched (HUFA-enriched) fish oil] at three lipid levels (20, 60 and 100 g kg(-1) dry diet) for 8 weeks. Decreased feed intake, poor growth performance, hepatic pathology and higher blood lipid peroxidation were found in 60 and 100 g kg(-1) fish oil groups. Conversely, in lard and plant oil groups, even at 100 g kg(-1) dietary lipid level, feed intake and growth performance did not decrease, despite histological observation revealed hepatic pathology in these groups. Plasma triglyceride and cholesterol contents increased significantly in all 100 g kg(-1) dietary lipid groups. In the comparison of hepatic FA beta-oxidation among three oil groups at 60 g kg(-1) dietary lipid level, impaired mitochondrial and peroxisomal FA oxidation capacity was observed in fish oil group. The results confirmed the relatively low capacity of grass carp to utilize high dietary lipid, and furthermore excess HUFA intake will result in more serious adverse effects than other FA.

Daily exercise increases hepatic fatty acid oxidation and prevents steatosis in Otsuka Long-Evans Tokushima Fatty rats

Exercise training is commonly prescribed for treatment of nonalcoholic fatty liver disease (NAFLD). We sought to determine whether exercise training prevents the development of NAFLD in Otsuka Long-Evans Tokushima Fatty (OLETF) rats and to elucidate the molecular mechanisms underlying the effects of exercise on hepatic steatosis. Four-week-old OLETF rats were randomly assigned to either a sedentary control group (Sed) or a group given access to voluntary running wheels for 16 wk (Ex). Wheels were locked 2 days before euthanasia in the Ex animals, and both groups were euthanized at 20 wk old. Voluntary wheel running attenuated weight gain and reduced serum glucose, insulin, free fatty acids, and triglycerides in Ex animals compared with Sed (P < 0.001). Ex animals exhibited significantly reduced hepatic triglyceride levels and displayed fewer lipid droplets (Oil Red O staining) and reduced lipid droplet size compared with Sed. Wheel running increased by threefold the percent of palmitate oxidized completely to CO(2) in the Ex animals but did not alter AMP-activated protein kinase-alpha (AMPKalpha) or AMPK phosphorylation status. However, fatty acid synthase and acetyl-coenzyme A carboxylase (ACC) content were significantly reduced (approximately 70 and approximately 35%, respectively), and ACC phosphorylation and cytochrome c content were significantly elevated (approximately 35 and approximately 30%, respectively) in the Ex animals. These results unequivocally demonstrate that daily physical activity attenuates hepatic steatosis and NAFLD in an obese rodent model and suggest that this effect is likely mediated, in part, through enhancement of hepatic fatty acid oxidation and reductions in key protein intermediates of fatty acid synthesis.

Rat liver mitochondrial carnitine palmitoyltransferase-I, hepatic carnitine, and malonyl-CoA: Effect of starvation

Hepatic mitochondrial fatty acid oxidation and ketogenesis increase during starvation. Carnitine palmitoyltransferase I (CPT-I) catalyses the rate-controlling step in the overall pathway and retains its control over β-oxidation under fed, starved and diabetic conditions. To determine the factors contributing to the reported several-fold increase in fatty acid oxidation in perfused livers, we measured the Vmax and Km values for palmitoyl-CoA and carnitine, the Ki (and IC50) values for malonyl-CoA in isolated liver mitochondria as well as the hepatic malonyl-CoA and carnitine contents in control and 48 h starved rats. Since CPT-I is localized in the mitochondrial outer membrane and in contact sites, the kinetic properties of CPT-I also was determined in these submitochondrial structures. After 48 h starvation, there is: (a) a significant increase in Ki and decrease in hepatic malonyl-CoA content; (b) a decreased Km for palmitoyl-CoA; and (c) increased catalytic activity (Vmax) and CPT-I protein abundance that is significantly greater in contact sites compared with outer membranes. Based on these changes the estimated increase in mitochondrial fatty acid oxidation is significantly less than that observed in perfused liver. This suggests that CPT-I is regulated in vivo by additional mechanism(s) lost during mitochondrial isolation or/and that mitochondrial oxidation of peroxisomal β-oxidation products contribute to the increased ketogenesis by bypassing CPT-I. Furthermore, the greater increase in CPT-I protein in contact sites as compared to outer membranes emphasizes the significance of contact sites in hepatic fatty acid oxidation.

904 IMIDAZOLYL DERIVATIVES IMPROVED RAT ALCOHOLIC (ASH) AND NON-ALCOHOLIC (NASH) STEATOHEPATITIS INDUCED BY LIQUID DIETS

In our recently published study we have detected in livers from SpragueDawley rats given fructose-enriched diet (FED) and developed NAFLD, increased hepatic triglycerides (+198%), increased hepatic cholesterol (+89%) and hypertriglyceridemia (+223%). The genetic risk factors involved in the pathogenesis of NAFLD are poorly understood. Methods: We investigated the gene expressions of: 1. Microsomal triglyceride transfer protein (MTP) involved in hepatic lipid synthesis, storage and export. 2. Manganese-dependent superoxide dismutase 2 (MnSOD2), encoding proteins involved in the response to oxidant stress. 3. Peroxisome proliferator-activated receptor (PPAR)-a involved in hepatic fatty acid oxidation, in rats that were administered FED (n = 6) or standard rat chow (n = 3) for five weeks. Intrahepatic messenger RNA (mRNA) levels of MTP, MnSOD2 and PPAR-a were measured by real-time polymerase chain reaction using Taqman gene expression assays (Applied Biosystems, Foster City, CA) in FED rats and in standard rat chow. Results: In the FED rats, liver histology demonstrated diffused macrovesicular and micro-vesicular steatosis. 1. MTP mRNA level was statistically significant decreased in FED rats (by 85%) (P< 0.0001) compared with the standard rat chow. 2. MnSOD2 mRNA level was statistically significant decreased in FED rats (by 80%) (P< 0.001) compared with the standard rat chow. 3. PPAR-a mRNA level was statistically significant decreased in FED rats (by 95%) (P< 0.0001) compared with the standard rat chow. Conclusions: the reduced expression of MTP, MnSOD2 and PPAR-a in the FED rats implies these genes are involved in the pathogenesis of NAFLD. Further studies are needed to confirm our results.

Alterations in CPT‐1 mRNA and fatty acid profile in hepatic cell lines in response to treatment with t10,c12‐ or c9,t11‐conjugated linoleic acid

AbstractTrans10,cis12‐conjugated linoleic acid (t10,c12‐CLA) increases liver weights and hepatic lipids in mice. The purpose of this study was to determine the effects of CLA isomers (t10,c12‐CLA or c9,t11‐CLA) and carnitine palmitoyl transferase‐1 (CPT‐1) inhibitors (etomoxir or hemipalmitoylcarnitinium) on CPT‐1 mRNA, fatty acid profile, and cholesterol synthesis in AML‐12 and HepG2 cells. t10,c12‐CLA was incorporated to a greater extent in both cell lines than c9,t11‐CLA. In addition, t10,c12‐CLA increased the free cholesterol content of AML‐12 and HepG2 cells four‐ and fivefold, respectively. Cells incubated with medium containing CPT‐1 inhibitors or t10,c12‐CLA had higher levels of mRNA for CPT‐1 in both cell lines, indicating an increased fatty acid oxidation in hepatic cell lines due to t10,c12‐CLA. Following treatment withdrawal, percentages of c9,t11‐CLA or t10,c12‐CLA remained elevated in cells initially treated with c9,t11‐CLA or t10,c12‐CLA, suggesting a potential for carryover effects of the CLA isomers. The results presented here demonstrate a potential role for t10,c12‐CLA in the modulation of hepatic fatty acid oxidation and cholesterol synthesis.

Hypertriglyceridemia and Hepatic Steatosis in Senescence-Accelerated Mouse Associate to Changes in Lipid-Related Gene Expression

Aged rodents show increasing plasma and tissue triglycerides, and reductions in liver peroxisome proliferator-activated receptor alpha (PPARalpha) and its target genes. We determined whether a similar situation is present in a model of accelerated aging, the senescence-accelerated prone (SAM-P8) mouse. Five-month-old SAM-P8 mice were hypertriglyceridemic, and exhibited hepatic steatosis and reduced fatty acid oxidation versus control 5-month-old senescence-accelerated resistant (SAM-R1) mice, with no differences in PPARalpha expression and binding activity; in fact, fenofibrate administration to SAM-P8 mice induced a clear PPARalpha-driven response. Complementary DNA (cDNA) microarray analysis (Affymetrix Mouse Genome 430A 2.0 GeneChip array), Western blot, and electrophoretic mobility shift assay (EMSA) experiments indicated, among other changes, a deficit in farnesoid X receptor (FXR) expression and binding activity in the livers of SAM-P8 mice with respect to SAM-R1 controls. Triglyceride accretion and a deficit in hepatic fatty acid oxidation, features of the aging process in mammals, associate to a deficit in hepatic FXR activity in the SAM-P8 mice.

Fenofibrate modulates cardiac and hepatic metabolism and increases ischemic tolerance in diet-induced obese mice

Peroxisome proliferator-activated receptors (PPARs) play an important role in the transcriptional regulation of lipid utilization and storage in several organs, including liver and heart. Our working hypothesis is that treatment of obesity/hyperlipedemia with the PPARα ligand fenofibrate leads to drainage of plasma lipids by the liver, resulting in reduced myocardial lipid supply, reduced myocardial fatty acid oxidation and improved myocardial tolerance to ischemic stress. Thus, we investigated changes in substrate utilization in heart and liver, as well as post-ischemic functional recovery in hearts from diet-induced obese (DIO) mice following long-term (11–12 weeks) treatment with fenofibrate. The present study shows that DIO mice express increased plasma lipids and glucose, as well as increased myocardial fatty acid oxidation and a concomitant decrease in glucose oxidation. The lipid-lowering effect of fenofibrate was associated with increased hepatic mitochondrial and peroxisomal fatty acid oxidation, as indicated by a more than 30% increase in hepatic palmiotyl-CoA oxidation and more than a 10-fold increase in acyl-CoA oxidase (ACO) activity. In line with an adaptation to the reduced myocardial lipid supply, isolated hearts from fenofibrate-treated DIO mice showed increased glucose oxidation and decreased fatty acid oxidation, as well as reduced ACO activity. Fenofibrate treatment also prevented the diet-induced decrease in cardiac function and improved post-ischemic functional recovery. We also found that, while fenofibrate treatment markedly increased the expression of PPARα target genes in the liver, there were no such changes in the heart. These data demonstrate that fenofibrate results in a direct activation of PPARα in the liver with increased hepatic drainage of plasma lipids, while the cardiac effect of the compound most likely is secondary to its lipid-lowering effect.

Suppression of Diacylglycerol Acyltransferase-2 (DGAT2), but Not DGAT1, with Antisense Oligonucleotides Reverses Diet-induced Hepatic Steatosis and Insulin Resistance

Nonalcoholic fatty liver disease (NAFLD) is a major contributing factor to hepatic insulin resistance in type 2 diabetes. Diacylglycerol acyltransferase (Dgat), of which there are two isoforms (Dgat1 and Dgat2), catalyzes the final step in triglyceride synthesis. We evaluated the metabolic impact of pharmacological reduction of DGAT1 and -2 expression in liver and fat using antisense oligonucleotides (ASOs) in rats with diet-induced NAFLD. Dgat1 and Dgat2 ASO treatment selectively reduced DGAT1 and DGAT2 mRNA levels in liver and fat, but only Dgat2 ASO treatment significantly reduced hepatic lipids (diacylglycerol and triglyceride but not long chain acyl CoAs) and improved hepatic insulin sensitivity. Because Dgat catalyzes triglyceride synthesis from diacylglycerol, and because we have hypothesized that diacylglycerol accumulation triggers fat-induced hepatic insulin resistance through protein kinase C epsilon activation, we next sought to understand the paradoxical reduction in diacylglycerol in Dgat2 ASO-treated rats. Within 3 days of starting Dgat2 ASO therapy in high fat-fed rats, plasma fatty acids increased, whereas hepatic lysophosphatidic acid and diacylglycerol levels were similar to those of control rats. These changes were associated with reduced expression of lipogenic genes (SREBP1c, ACC1, SCD1, and mtGPAT) and increased expression of oxidative/thermogenic genes (CPT1 and UCP2). Taken together, these data suggest that knocking down Dgat2 protects against fat-induced hepatic insulin resistance by paradoxically lowering hepatic diacylglycerol content and protein kinase C epsilon activation through decreased SREBP1c-mediated lipogenesis and increased hepatic fatty acid oxidation.

Fasting-dependent glucose and lipid metabolic response through hepatic sirtuin 1

In the fasted state, induction of hepatic glucose output and fatty acid oxidation is essential to sustain energetic balance. Production and oxidation of glucose and fatty acids by the liver are controlled through a complex network of transcriptional regulators. Among them, the transcriptional coactivator PGC-1alpha plays an important role in hepatic and systemic glucose and lipid metabolism. We have previously demonstrated that sirtuin 1 (SIRT1) regulates genes involved in gluconeogenesis through interaction and deacetylation of PGC-1alpha. Here, we show in vivo that hepatic SIRT1 is a factor in systemic and hepatic glucose, lipid, and cholesterol homeostasis. Knockdown of SIRT1 in liver caused mild hypoglycemia, increased systemic glucose and insulin sensitivity, and decreased glucose production. SIRT1 knockdown also decreased serum cholesterol and increased hepatic free fatty acid and cholesterol content. These metabolic phenotypes caused by SIRT1 knockdown tightly correlated with decreased expression of gluconeogenic, fatty acid oxidation and cholesterol degradation as well as efflux genes. Additionally, overexpression of SIRT1 reversed many of the changes caused by SIRT1 knockdown and depended on the presence of PGC-1alpha. Interestingly, most of the effects of SIRT1 were only apparent in the fasted state. Our results indicate that hepatic SIRT1 is an important factor in the regulation of glucose and lipid metabolism in response to nutrient deprivation. As these pathways are dysregulated in metabolic diseases, SIRT1 may be a potential therapeutic target to control hyperglycemia and hypercholesterolemia.

Cideb Regulates Diet-Induced Obesity, Liver Steatosis, and Insulin Sensitivity by Controlling Lipogenesis and Fatty Acid Oxidation

Our previous study suggests that Cidea, a member of Cide family proteins that share sequence homology with the DNA fragmentation factor and are expressed at high levels in brown adipose tissue, plays an important role in the development of obesity. Cideb, another member of Cide family protein, is highly expressed in the liver. We would like to understand the physiological role of Cideb in the regulation of energy expenditure and lipid metabolism. We generated Cideb-null mice by homolog recombination and then fed both wild-type and Cideb-null mice with high-fat diet (58% fat). We then characterized the animals' adiposity index, food intake, whole-body metabolic rate, liver morphology, rate of fatty acid synthesis and oxidation, insulin sensitivity, and gene expression profile. Cideb-null mice had lower levels of plasma triglycerides and free fatty acids and were resistant to high-fat diet-induced obesity and live steatosis. In addition, Cideb mutant mice displayed significantly increased insulin sensitivity and enhanced rate of whole-body metabolism and hepatic fatty acid oxidation. More importantly, Cideb-null mice showed decreased lipogenesis and reduced expression levels of acetyl-CoA carboxylase, fatty acid synthase, and stearol-CoA desaturase. We further demonstrated that expression levels of sterol response element binding protein 1c was significantly decreased in Cideb-deficient mice. Our data demonstrate that Cideb is a novel important regulator in lipid metabolism in the liver. Cideb may represent a new therapeutic target for the treatment of obesity, diabetes, and liver steatosis.

Mercaptoacetate's feeding stimulatory effect is not increased in rats fed a medium-chain triglyceride-rich diet.

Intraperitoneal (ip) injection of mercaptoacetate (MA) inhibits hepatic fatty acid oxidation (FAO) and stimulates feeding in rats fed diets containing substantial amounts of lipid. Because it remains unclear if the reduction in hepatic FAO causes the increase in feeding, we compared the effects of MA (45.6 mg/kg BW; injected ip at light onset) vs. saline control in rats adapted to diets containing 18% either predominately long-chain (about 95% 16 C and more) or medium-chain (about 70% 10C–14C) fatty acid triglycerides (LCT and MCT, respectively). If MA's feeding stimulatory effect depends on FAO, MA should stimulate feeding more in MCT-fed rats because medium-chain fatty acids are oxidized faster than long-chain fatty acids. To assess) possible metabolic effects of the diets and of MA, rats were killed 30 min after light-onset injections for blood and tissue (liver, duodenum) samples. Relative concentrations of fatty acids in the duodenal tissue matched the patterns in the diets fed. Two results suggested that hepatic FAO proceeded more rapidly in MCT-fed rats. First, plasma β-hydroxybutyrate levels were higher in MCT—than in LCT-fed rats. Second, the relative hepatic content of fatty acids in the liver compared to the diet was reduced more for medium-chain fatty acids than for long-chain fatty acids. Contrary to our hypothesis, however, MA increased 1-h food intake more in LCT than MCT-fed rats (290% vs. 54%, p