The miniature pig is a very suitable donor species in xenotransplantation of human organs. Lipid metabolism is an important process that involves the creation and degradation of lipids, which is associated with the function of the gastro-intestinal tract. However, the distribution of lipid metabolism related molecules in the gastro-intestinal tract in the miniature pig is unclear. The present study examined the expression of farnesoid X-receptor (FXR), liver X- receptor (LXR), retinoid X-receptor (RXR), liver fatty acid binding protein (L-FABP), fatty acid synthase (FAS) mRNA in the digestive organs of miniature pigs. FXR and LXR mRNA were not expressed in the stomach but were expressed at high and low density in the small and large intestines, respectively. RXR mRNA was expressed in stomach with moderate density, small intestine with high density and in the large intestine with low density. L-FABP and FAS mRNA were expressed in the stomach and large intestine with low density and in the small intestine with high density. L-FABP mRNA was expressed in the liver and kidney with high density, and in pancreas with low density. FAS mRNA was expressed in the liver with high density, and in pancreas and kidney with low density.
Fatty Acid Synthetase Complex ; Fatty Acid-Binding Proteins ; Humans ; Intestine, Large ; Intestine, Small ; Intestines ; Kidney ; Lipid Metabolism ; Liver ; Pancreas ; RNA, Messenger ; Stomach ; Swine ; Tissue Donors ; Transplantation, Heterologous

The miniature pig is a very suitable donor species in xenotransplantation of human organs. Lipid metabolism is an important process that involves the creation and degradation of lipids, which is associated with the function of the gastro-intestinal tract. However, the distribution of lipid metabolism related molecules in the gastro-intestinal tract in the miniature pig is unclear. The present study examined the expression of farnesoid X-receptor (FXR), liver X- receptor (LXR), retinoid X-receptor (RXR), liver fatty acid binding protein (L-FABP), fatty acid synthase (FAS) mRNA in the digestive organs of miniature pigs. FXR and LXR mRNA were not expressed in the stomach but were expressed at high and low density in the small and large intestines, respectively. RXR mRNA was expressed in stomach with moderate density, small intestine with high density and in the large intestine with low density. L-FABP and FAS mRNA were expressed in the stomach and large intestine with low density and in the small intestine with high density. L-FABP mRNA was expressed in the liver and kidney with high density, and in pancreas with low density. FAS mRNA was expressed in the liver with high density, and in pancreas and kidney with low density.
Fatty Acid Synthetase Complex ; Fatty Acid-Binding Proteins ; Humans ; Intestine, Large ; Intestine, Small ; Intestines ; Kidney ; Lipid Metabolism ; Liver ; Pancreas ; RNA, Messenger ; Stomach ; Swine ; Tissue Donors ; Transplantation, Heterologous

BACKGROUND: Amniotic fluid is a rich source of fetal mesenchymal stem cells (MSCs). However, little is known about whether bisphosphonates affect the differentiation into adipocytes. Therefore, this study was aimed to investigate whether zoledronate influences the differentiation of AFMSCs into adipocytes. METHODS: Amniotic fluid cells samples were obtained from 6 pregnant women by second trimester amniocentesis for performing fetal karyotyping. The cells were treated with various concentration (10(-10), 10(-8), 10(-6) M) of zoledronate and the cells were analyzed over 21 days of culture. Differentiation into adipocytes was determined by oil-red O staining and for fatty acid synthase (FAS), acetyl CoA carboxylase 1 (ACC1) and sterol regulatory elementary binding protein-1 (SREBP-1). RESULTS: Differentiation of AFMSCs into adipocytes was found by oil-red O staining. Zoledronate influenced the differentiation of AFMSCs into adipocytes in a dose- and time-dependent manner. At 7 days of culture, the expressions of FAS and SREBP-1 showed no significant differences compared to that of the control regardless of the dose of zoledronate. Very little ACC1 expression was found. However, the expressions of these three markers were remarkably increased at 14 days of culture. Of them, the ACC1 expression was significantly increased by 10(-8) M and 10(-6) M of zoledronate. At 21 days of culture, there were no effects of zoledronate on the expressions of FAS and SREBP-1. However, the ACC1 expression was decreased with an increasing dose of zoledronate (P<.05). CONCLUSION: This study shows that AFMSCs can be differentiated into adipocytes. The induction of this differentiation following zoledronate treatment appears to be dose dependent and time-of-culture dependent.
Acetyl-CoA Carboxylase ; Adipocytes ; Amniocentesis ; Amniotic Fluid ; Diphosphonates ; Fatty Acid Synthetase Complex ; Female ; Humans ; Imidazoles ; Karyotyping ; Mesenchymal Stromal Cells ; Pregnancy ; Pregnancy Trimester, Second ; Pregnant Women ; Stem Cells

BACKGROUND: Insulin resistance is very common in patients with nonalcoholic fatty liver disease (NAFLD). Glitazones improve insulin sensitivity by acting as a selective agonist of the peroxisome proliferators -activated receptor gamma (PPAR gamma), and were shown to activate AMP-activated protein kinase (AMPK) in skeletal muscle and the liver. Glitazones were also shown to reduce hepatic lipogenesis. The aim of this study was to investigate whether the protective mechanism of rosiglitazone on NAFLD is associated with AMPK activation. METHODS: Twelve OLETF rats were divided into 2 groups (control, treatment, n = 6 each). LETO rats served as controls. At 35 weeks of age, treatment group received rosiglitazone 4 mg/kg daily for 3 days. Fasting plasma glucose, insulin, free fatty acid, lactate and triglycerides were measured. Liver tissues from each group were processed for histological and hepatic triglyceride content analysis and western blotting. RESULTS: Fasting plasma glucose, insulin and triglycerides levels were significantly lower in treatment group than in control group. Histologic examination disclosed decreased hepatic steatosis in treatment group. Hepatic triglyceride content was also decreased in treatment group. Sterol regulatory binding protein-1c (SREBP-1c) and fatty acid synthase (FAS) expression were increased and AMPK phosphorylation was reduced in OLETF rats compared with LETO rats, and these changes were reversed by rosiglitazone treatment. CONCLUSION: Rosiglitazone reduced hepatic steatosis in OLETF rats, and activated AMPK in the liver. These results suggest the role of AMPK activation in the protective action of rosiglitazone on NAFLD.
AMP-Activated Protein Kinases ; Animals ; Fasting ; Fatty Acid Synthetase Complex ; Fatty Liver ; Glucose ; Humans ; Insulin ; Insulin Resistance ; Lactic Acid ; Lipogenesis ; Liver ; Muscle, Skeletal ; Peroxisome Proliferators ; Phosphorylation ; Plasma ; Rats ; Rats, Inbred OLETF ; Thiazolidinediones ; Triglycerides

The regulation of fatty acid synthase in rat liver was investigated at transcriptional and post-transcriptional levels. When rats were fasted for 3 days and refed on a high-carbohydrate diet, the amounts of FAS in liver cytosol began to increase at 12 hours and further increased until 48 hours. The amount of mRNA for FAS began to increase at 6 hours and reached to a maximum level at 12 hours, indicating that the expression of mRNA for FAS precedes the increase of FAS protein pool. After 12 hours the amounts of mRNA gradually decreased and remained at a much lowered level between 24 and 48 hours. The elevated amount of FAS mRNA reflected on the amount of FAS protein in the first 24 hours, but these two parameters were not paralleled thereafter, probably due to the changes in the translational efficiencies. The run-on transcriptional activity of FAS gene began to increase at 4 hours after refeeding a high-carbohydrate diet and further increased to reach a maximum level 25 fold of the initial level at 12 hours, followed by a 16 fold level between 24 and 48 hours. The elevation of run-on transcriptional activity of FAS gene preceded the increase of FAS mRNA in the liver cytosol by 2 hours, and a similar increasing pattern was observed until 12 hours. However, FAS mRNA concentration decreased gradually after 12 hours, while the transcriptional activity remained at a high level until 48 hours. The changes in FAS mRNA content in the cytosol of rat liver were closely related to the transcriptional activity of FAS gene in the early phase of induction, but another regulatory mechanism seems to operate in the decrease of mRNA after 12 hours.
Animal ; DNA/genetics ; Dietary Carbohydrates/administration & dosage ; Fatty Acid Synthetase Complex/*biosynthesis/genetics ; *Gene Expression Regulation, Enzymologic ; Liver/*enzymology ; Male ; RNA, Messenger/metabolism ; Rats ; Rats, Sprague-Dawley ; *Transcription, Genetic

BACKGROUND: S-methylmethionine sulfonium chloride was originally called vitamin U because of its inhibition of ulceration in the digestive system. Vitamin U is ubiquitously expressed in the tissues of flowering plants, and while there have been reports on its hypolipidemic effect, its precise function remains unknown. OBJECTIVE: This study was designed to evaluate the anti-obesity effect of vitamin U in 3T3-L1 pre-adipocyte cell lines. METHODS: We cultured the pre-adipocyte cell line 3T3L1 to overconfluency and then added fat differentiation-inducing media (dexamethasone, IBMX [isobutylmethylxanthine], insulin, indomethacin) and different concentrations (10, 50, 70, 90, 100 mM) of vitamin U. Then, we evaluated changes in the levels of triglycerides (TGs), glycerol-3-phosphate dehydrogenase (G3PDH), AMP-activated protein kinase (AMPK), adipocyte-specific markers (peroxisome proliferator-activated receptor gamma [PPAR-gamma], CCAAT/enhancer-binding protein alpha [C/EBP-alpha], adipocyte differentiation and determination factor 1 [ADD-1], adipsin, fatty acid synthase, lipoprotein lipase) and apoptosis-related signals (Bcl-2, Bax). RESULTS: There was a gradual decrease in the level of TGs, C/EBP-alpha, PPAR-gamma, adipsin, ADD-1 and GPDH activity with increasing concentrations of vitamin U. In contrast, we observed a significant increase in AMPK activity with increasing levels of vitamin U. The decrease in bcl-2 and increase in Bax observed with increasing concentrations of vitamin U in the media were not statistically significant. CONCLUSION: This study suggests that vitamin U inhibits adipocyte differentiation via down-regulation of adipogenic factors and up-regulation of AMPK activity.
1-Methyl-3-isobutylxanthine ; Adipocytes ; AMP-Activated Protein Kinases ; Cell Line ; Complement Factor D ; Digestive System ; Down-Regulation ; Fatty Acid Synthetase Complex ; Flowers ; Glycerolphosphate Dehydrogenase ; Insulin ; Lipoproteins ; Triglycerides ; Ulcer ; Up-Regulation ; Vitamin U ; Vitamins

This study was conducted to investigate the effects of powdered young barley leaf and its water extract on body weight and lipid metabolism in high-fat fed mice. Male mice were divided into normal group, high-fat (HF) group, highfat group supplemented with powdered young barley leaf (HF-YBL) and high-fat group supplemented with water extract of the powdered young barley leaf (HF-WYBL). The powdered young barley leaf or its water extract was added to a standard diet based on 1% dried young barley leaf (1 g YBL/100 diet and 0.28 g WYBL/100 g diet) for 8 weeks. Supplementation of YBL and WYBL significantly reduced body weight and epididymal adipose tissue weight in highfat fed mice. Food intake and daily energy intake were significantly lower in the YBL group than in the HF group. After 8 weeks, plasma triglyceride and cholesterol concentrations were significantly higher in the HF group than in the Normal group; however, both YBL and WYBL significantly lowered those of the high-fat fed mice. The ratio of HDL-cholesterol/ total cholesterol of the YBL and WYBL groups were significantly elevated compared to that of HF group. Both YBL and WYBL significantly increased fecal excretion of triglyceride in high-fat fed mice, whereas they did not affect fecal cholesterol concentration. The triglyceride levels of liver, adipose tissue and heart were significantly lower in the YBL and WYBL groups than in the HF group. Supplementation of WYBL also lowered the kidney triglyceride and heart cholesterol concentrations compared to those of HF group. Hepatic lipid regulating enzyme activities, fatty acid synthase, HMG-CoA reductase and acyl-coenzyme A: cholesterol acyltransferase, were significantly lower in the YBL and WYBL groups than in the HF group. Accordingly, these results suggest that YBL and WYBL improve plasma and organ lipid levels partly by increasing fecal lipid excretion and inhibiting fatty acid and cholesterol biosynthesis in the liver.
Acyl Coenzyme A ; Adipose Tissue ; Animals ; Body Weight ; Cholesterol ; Diet ; Diet, High-Fat ; Eating ; Energy Intake ; Fatty Acid Synthetase Complex ; Heart ; Hordeum ; Humans ; Kidney ; Lipid Metabolism ; Liver ; Male ; Mice ; Oxidoreductases ; Plasma ; Sterol O-Acyltransferase ; Water

In this study, we examined the hepatic anti-steatosis activity of carnosic acid (CA), a phenolic compound of rosemary (Rosmarinus officinalis) leaves, as well as its possible mechanism of action, in a high-fat diet (HFD)-fed mice model. Mice were fed a HFD, or a HFD supplemented with 0.01% (w/w) CA or 0.02% (w/w) CA, for a period of 12 weeks, after which changes in body weight, blood lipid profiles, and fatty acid mechanism markers were evaluated. The 0.02% (w/w) CA diet resulted in a marked decline in steatosis grade, as well as in homeostasis model assessment of insulin resistance (HOMA-IR) index values, intraperitoneal glucose tolerance test (IGTT) results, body weight gain, liver weight, and blood lipid levels (P < 0.05). The expression level of hepatic lipogenic genes, such as sterol regulating element binding protein-1c (SREBP-1c), liver-fatty acid binding protein (L-FABP), stearoyl-CoA desaturase 1 (SCD1), and fatty acid synthase (FAS), was significantly lower in mice fed 0.01% (w/w) CA and 0.02% (w/w) CA diets than that in the HFD group; on the other hand, the expression level of beta-oxidation-related genes, such as peroxisome proliferator-activated receptor alpha (PPAR-alpha), carnitine palmitoyltransferase 1 (CPT-1), and acyl-CoA oxidase (ACO), was higher in mice fed a 0.02% (w/w) CA diet, than that in the HFD group (P < 0.05). In addition, the hepatic content of palmitic acid (C16:0), palmitoleic acid (C16:1), and oleic acid (C18:1) was significantly lower in mice fed the 0.02% (w/w) CA diet than that in the HFD group (P < 0.05). These results suggest that orally administered CA suppressed HFD-induced hepatic steatosis and fatty liver-related metabolic disorders through decrease of de novo lipogenesis and fatty acid elongation and increase of fatty acid beta-oxidation in mice.
Acyl Coenzyme A ; Acyl-CoA Oxidase ; Animals ; Body Weight ; Carnitine O-Palmitoyltransferase ; Carrier Proteins ; Diet ; Diet, High-Fat ; Diterpenes, Abietane ; Fatty Acid Synthetase Complex ; Fatty Acids, Monounsaturated ; Glucose Tolerance Test ; Hand ; Homeostasis ; Insulin Resistance ; Lipogenesis ; Liver ; Mice ; Oleic Acid ; Palmitic Acid ; Phenol ; Plant Extracts ; PPAR alpha ; Stearoyl-CoA Desaturase

BACKGROUND: Enhanced lipogenesis plays a critical role in cell senescence via induction of expression of the mature form of sterol regulatory element binding protein 1 (SREBP1), which contributes to an increase in organellar mass, one of the indicators of senescence. We investigated the molecular mechanisms by which signaling molecules control SREBP1-mediated lipogenesis and senescence. METHODS: We developed cellular models for stress-induced senescence, by exposing Chang cells, which are immortalized human liver cells, to subcytotoxic concentrations (200 microM) of deferoxamine (DFO) and H2O2. RESULTS: In this model of stress-induced cell senescence using DFO and H2O2, the phosphorylation profile of glycogen synthase kinase 3alpha (GSK3alpha) and beta corresponded closely to the expression profile of the mature form of SREBP-1 protein. Inhibition of GSK3 with a subcytotoxic concentration of the selective GSK3 inhibitor SB415286 significantly increased mature SREBP1 expression, as well as lipogenesis and organellar mass. In addition, GSK3 inhibition was sufficient to induce senescence in Chang cells. Suppression of GSK3 expression with siRNAs specific to GSK3alpha and beta also increased mature SREBP1 expression and induced senescence. Finally, blocking lipogenesis with fatty acid synthase inhibitors (cerulenin and C75) and siRNA-mediated silencing of SREBP1 and ATP citrate lyase (ACL) significantly attenuated GSK3 inhibition-induced senescence. CONCLUSION: GSK3 inactivation is an important upstream event that induces SREBP1-mediated lipogenesis and consequent cell senescence.
Aging* ; Aminophenols ; ATP Citrate (pro-S)-Lyase ; Carrier Proteins* ; Cell Aging ; Deferoxamine ; Fatty Acid Synthetase Complex ; Glycogen Synthase Kinase 3* ; Glycogen Synthase Kinases* ; Glycogen Synthase* ; Glycogen* ; Humans ; Lipogenesis* ; Liver ; Maleimides ; Multienzyme Complexes ; Oxo-Acid-Lyases ; Phosphorylation ; RNA, Small Interfering ; Sterol Regulatory Element Binding Protein 1

BACKGROUND: Enhanced lipogenesis plays a critical role in cell senescence via induction of expression of the mature form of sterol regulatory element binding protein 1 (SREBP1), which contributes to an increase in organellar mass, one of the indicators of senescence. We investigated the molecular mechanisms by which signaling molecules control SREBP1-mediated lipogenesis and senescence. METHODS: We developed cellular models for stress-induced senescence, by exposing Chang cells, which are immortalized human liver cells, to subcytotoxic concentrations (200 microM) of deferoxamine (DFO) and H2O2. RESULTS: In this model of stress-induced cell senescence using DFO and H2O2, the phosphorylation profile of glycogen synthase kinase 3alpha (GSK3alpha) and beta corresponded closely to the expression profile of the mature form of SREBP-1 protein. Inhibition of GSK3 with a subcytotoxic concentration of the selective GSK3 inhibitor SB415286 significantly increased mature SREBP1 expression, as well as lipogenesis and organellar mass. In addition, GSK3 inhibition was sufficient to induce senescence in Chang cells. Suppression of GSK3 expression with siRNAs specific to GSK3alpha and beta also increased mature SREBP1 expression and induced senescence. Finally, blocking lipogenesis with fatty acid synthase inhibitors (cerulenin and C75) and siRNA-mediated silencing of SREBP1 and ATP citrate lyase (ACL) significantly attenuated GSK3 inhibition-induced senescence. CONCLUSION: GSK3 inactivation is an important upstream event that induces SREBP1-mediated lipogenesis and consequent cell senescence.
Aging* ; Aminophenols ; ATP Citrate (pro-S)-Lyase ; Carrier Proteins* ; Cell Aging ; Deferoxamine ; Fatty Acid Synthetase Complex ; Glycogen Synthase Kinase 3* ; Glycogen Synthase Kinases* ; Glycogen Synthase* ; Glycogen* ; Humans ; Lipogenesis* ; Liver ; Maleimides ; Multienzyme Complexes ; Oxo-Acid-Lyases ; Phosphorylation ; RNA, Small Interfering ; Sterol Regulatory Element Binding Protein 1