Fatty Acid Synthase Gene Expression Research Articles

2-Deoxy-D-Glucose Downregulates Fatty Acid Synthase Gene Expression Via an Endoplasmic Reticulum Stress-Dependent Pathway in HeLa Cells.

Fatty acid synthase (FASN) catalyzes the rate-limiting step of cellular lipogenesis. FASN expression is upregulated in various types of cancer cells, implying that FASN is a potential target for cancer therapy. 2-Deoxy-D-glucose (2-DG) specifically targets cancer cells by inhibiting glycolysis and glucose metabolism, resulting in multiple anticancer effects. However, whether the effects of 2-DG involve lipogenic metabolism remains to be elucidated. We investigated the effect of 2-DG administration on FASN expression in HeLa human cervical cancer cells. 2-DG treatment for 24 h decreased FASN mRNA and protein levels and suppressed the activity of an exogenous rat Fasn promoter. The use of a chemical activator or inhibitors or of a mammalian expression plasmid showed that neither AMPK nor the Sp1 transcription factor is responsible for the inhibitory effect of 2-DG on FASN expression. Administration of thapsigargin, an endoplasmic reticulum (ER) stress inducer, or 4-(2-aminoethyl) benzenesulfonyl fluoride (AEBSF), a site 1 protease inhibitor, mimicked the inhibitory effect of 2-DG on FASN expression. 2-DG did not further decrease FASN expression in the presence of thapsigargin or AEBSF. Site 1 protease mediates activation of ATF6, an ER stress mediator, as well as sterol regulatory element-binding protein 1 (SREBP1), a robust transcription factor for FASN. Administration of 2-DG or thapsigargin for 24 h suppressed activation of ATF6 and SREBP1, as did AEBSF. We speculated that these effects of 2-DG or thapsigargin are due to feedback inhibition via increased GRP78 expression following ER stress. Supporting this, exogenous overexpression of GRP78 in HeLa cells suppressed SREBP1 activation and Fasn promoter activity. These results suggest that 2-DG suppresses FASN expression via an ER stress-dependent pathway, providing new insight into the molecular basis of FASN regulation in cancer.

Mitochondrial Genome Editing: Exploring the Possible Relationship of the Atherosclerosis-Associated Mutation m.15059G>A With Defective Mitophagy.

The aim of this study was to evaluate the effect of the m.15059G>A mitochondrial nonsense mutation on cellular functions related to atherosclerosis, such as lipidosis, pro-inflammatory response, and mitophagy. Heteroplasmic mutations have been proposed as a potential cause of mitochondrial dysfunction, potentially disrupting the innate immune response and contributing to the chronic inflammation associated with atherosclerosis. The human monocytic cell line THP-1 and cytoplasmic hybrid cell line TC-HSMAM1 were used. An original approach based on the CRISPR/Cas9 system was developed and used to eliminate mitochondrial DNA (mtDNA) copies carrying the m.15059G>A mutation in the MT-CYB gene. The expression levels of genes encoding enzymes related to cholesterol metabolism were analyzed using quantitative polymerase chain reaction. Pro-inflammatory cytokine secretion was assessed using enzyme-linked immunosorbent assays. Mitophagy in cells was detected using confocal microscopy. In contrast to intact TC-HSMAM1 cybrids, Cas9-TC-HSMAM1 cells exhibited a decrease in fatty acid synthase (FASN) gene expression following incubation with atherogenic low-density lipoprotein. TC-HSMAM1 cybrids were found to have defective mitophagy and an inability to downregulate the production of pro-inflammatory cytokines (to establish immune tolerance) upon repeated lipopolysaccharide stimulation. Removal of mtDNA harboring the m.15059G>A mutation resulted in the re-establishment of immune tolerance and the activation of mitophagy in the cells under investigation. The m.15059G>A mutation was found to be associated with defective mitophagy, immune tolerance, and impaired metabolism of intracellular lipids due to upregulation of FASN in monocytes and macrophages.

Transcriptional and epigenetic changes after dietary and surgical weight loss interventions in an animal model of obesity.

Identifying determinants that can predict response to weight loss interventions is imperative for optimizing therapeutic benefit. We aimed to identify changes in DNA methylation and mRNA expression of a subset of target genes following dietary and surgical interventions in high-fat-diet (HFD)-induced obese rats. Forty-two adult Wistar Han male rats were divided into two groups: control rats (n = 7) and obese rats (n = 28), fed a HFD for 10 weeks (t10). Obese rats were randomly subdivided into five intervention groups (seven animals per group): (i) HFD; (ii) very-low-calorie diet (VLCD); (iii) sham surgery, and (iv) sleeve gastrectomy (SG). At week sixteen (t16), animals were sacrificed and tissue samples were collected to analyze changes in DNA methylation and mRNA expression of the selected genes. By type of intervention, the surgical procedures led to the greatest weight loss. Changes in methylation and/or expression of candidate genes occurred proportionally to the effectiveness of the weight loss interventions. Leptin expression, increased sixfold in the visceral fat of the obese rats, was partially normalized after all interventions. The expression of fatty acid synthase (FASN) and monocyte chemoattractant protein 1 (MCP-1) genes, which was reduced 0.5- and 0.15-fold, respectively, in the liver tissue of obese rats, were completely normalized after weight loss interventions, particularly after surgical interventions. The upregulation of FASN and MCP-1 gene expression was accompanied by a significant reduction in promoter methylation, up to 0.5-fold decrease in the case of the FASN (all intervention groups) and a 0.8-fold decrease in the case of the MCP-1 (SG group). Changes in tissue expression of specific genes involved in the pathophysiological mechanisms of obesity can be significantly attenuated following weight loss interventions, particularly surgery. Some of these genes are regulated by epigenetic mechanisms.

Coral calcium carried hydrogen ameliorates the severity of non-alcoholic steatohepatitis induced by a choline deficient high carbohydrate fat-free diet in elderly rats

Hydrogen has been reported to act as an antioxidant, anti-apoptosis and anti-inflammatory agent. Coral calcium carried hydrogen (G2-SUISO) is a safer and more convenient form of hydrogen agent than others. The mechanism underlying the hepatoprotective effects of G2-SUISO using an elderly non-alcoholic steatohepatitis (NASH) rat model was investigated. Two days after fasting, six-month-old elderly male F344/NSlc rats were given a choline deficient high carbohydrate fat-free (CDHCFF) diet from day 0 to day 3 as CDHCFF control group, and then switched to a normal diet from days 4 to 7 with or without 300 mg/kg G2-SUISO. Rats in each group were finally being sacrificed on day 3 or day 7. In the CDHCFF diet group, G2-SUISO decreased the liver weight-to-body weight ratio, the serum AST, ALT, total cholesterol levels, inflammatory infiltration, pro-inflammatory cytokine expression and lipid droplets with inhibiting lipogenic pathways by reducing sterol regulatory element-binding protein-1c, acetyl-CoA carboxylase and fatty acid synthase gene expression compared with the CDHCFF diet alone. G2-SUISO had beneficial effects of anti-apoptosis as well the down-regulation of pro-apoptotic molecules including NF-κB, caspase-3, caspase-9 and Bax. These findings suggest that G2-SUISO treatment exerts a significant hepatoprotective effect against steatosis, inflammation and apoptosis in elderly NASH rats.

Galbanic acid of Ferula assa-foetida L, as a regulator of the AMPK pathway in reduction of lipid accumulation in HepG2 cells

Introduction: Hepatic fat accumulation is a complication of non-alcoholic fatty liver disease (NAFLD). An AMP-activated protein kinase (AMPK) reduces the synthesis of fatty acids by inhibiting sterol regulatory element-binding transcription factor 1-c (SREBP1-C) and acetyl COA carboxylase (ACC). Objectives: We aimed to investigate the impress of galbanic acid (Gal) from the Ferula plant on AMPK and its inhibitory effect on lipogenic enzymes in HepG2 cells. Materials and Methods: In an applied-fundamental study, HepG2 cells were treated for 24 hours with Gal in palmitate (Pal). Resveratrol (RSV) was conducted as a positive control. Fatty acid synthase (FAS) and SREBP1-C gene expression were evaluated by reverse transcription polymerase chain reaction (RT-PCR). FAS, phospho-acetyl-CoA carboxylase (P-ACC), P-AMPK, AMPK, SREBP-1c, and ACC protein levels were measured by western blotting. Lipid accumulation was investigated qualitatively and semi-quantitatively with oil red. Results: The semi-quantitative results of oil revealed a substantial reduction (P<0.004) in lipid accumulation for treatment with Gal. The significant increase in the protein level of P-AMPK (P<0.001) and P-ACC (P=0.054) and significant decrease in FAS (P<0.003), SREBP-1c (P<0.001) and ACC (P<0.011) due to the effect galbanic acid was observed. FAS gene expression decreased significantly (P<0.009), while the decrease in SREBP-1c gene expression was not significant (P=0.303). Conclusion: These findings direct that galbanic acid can be a new regulator of AMPK. Hence, the present study may introduce galbanic acid as a new plan to positively regulate the AMPK pathway, which leads to the regulation of various cellular processes.

Dietary L-carnitine supplementation for heat stressed juvenile pearl gentian grouper(Epinephelusllanolatus (Epinephelus♂×Epinephelusfuscoguttatus♀): Effects on the antioxidant enzyme, survival, and gene expression

The effects of increasing dietary inclusion levels of L-carnitine at 0 (control group) 200, 400, 600, 1200, 1400, and 1600 mg/kg were evaluated in juvenile hybrid groupers exposed to heat shock stress. The feeding trial was conducted for eight weeks. After the feeding trial, hybrid groupers were subjected to a heat stress test for an hour at a temperature of 36 °C to determine the influence of the treatments on survival, biochemical parameters, and gene expression. The results of the current study showed that the survival rate of hybrid grouper was low in fish fed the control diet. A large number of survivors were observed in the treatment group with 400 mg/kg L-carnitine supplementation (LC 400), obtaining the highest survival rate. Heat stress decreased (P < 0.05) the activities of alkaline phosphatase and acid phosphatase in the control group but was increased in fish fed the LC 400 diet. Additionally, in the LC 400 treatment, malondialdehyde content decreased considerably (P < 0.05) compared to LC0. Hybrid grouper exposed to a high temperature of 36 °C for 1 h without L-carnitine (LC0, control group) supplementation showed the most increased Aspartate Aminotransferase activity (2.04 U L−1). Additionally, the LC 400 group reached the maximal value for the expressions of genes in the PPARs (peroxisome proliferator-activated receptors) signaling pathway, which showed a tendency of downregulation and, subsequently, up-regulation. The gene expression of FAS (fatty acid synthase) was discovered upstream and downstream (P < 0.05). The lowest mRNA level was found in the lowest L-carnitine supplementation treatment (200 mg/kg). A CPT-1 (carnitine palmitoyltransferase) that controls fatty acid oxidation showed upstream and downstream activity in response to increasing doses of L-carnitine supplementation. Our findings showed that dietary supplementation with L-carnitine 400 mg/kg reduced heat stress-related mortality and oxidative damage by enhancing the survival and oxidative enzymes of grouper, which provides a nutritional method of minimizing the consequences of heat exposure.

MicroRNA-199a-3p regulates proliferation and milk fat synthesis of ovine mammary epithelial cells by targeting VLDLR.

In our previous study, microRNA (miR)-199a-3p was found to be the most upregulated miRNA in mammary gland tissue during the non-lactation period compared with the peak-lactation period. However, there have been no reports describing the function of miR-199a-3p in ovine mammary epithelial cells (OMECs) and the biological mechanisms by which the miRNA affects cell proliferation and milk fat synthesis in sheep. In this study, the effect of miR-199a-3p on viability, proliferation, and milk fat synthesis of OMECs was investigated, and the target relationship of the miRNA with very low-density lipoprotein receptor (VLDLR) was also verified. Transfection with a miR-199a-3p mimic increased the viability of OMECs and the number of Edu-labeled positive OMECs. In contrast, a miR-199-3p inhibitor had the opposite effect with the miR-199a-3p mimic. The expression levels of three marker genes were also regulated by both the miR-199a-3p mimic and miR-199-3p inhibitor in OMECs. Together, these results suggest that miR-199a-3p promotes the viability and proliferation of OMECs. A dual luciferase assay confirmed that miR-199a-3p can target VLDLR by binding to the 3′-untranslated regions (3'UTR) of the gene. Further studies found a negative correlation in the expression of miR-199a-3p with VLDLR. The miR-199a-3p mimic decreased the content of triglycerides, as well as the expression levels of six milk fat synthesis marker genes in OMECs, namely, lipoprotein lipase gene (LPL), acetyl-CoA carboxylase alpha gene (ACACA), fatty acid binding protein 3 gene (FABP3), CD36, stearoyl-CoA desaturase gene (SCD), and fatty acid synthase gene (FASN). The inhibition of miR-199a-3p increased the level of triglycerides and the expression of LPL, ACACA, FABP3, SCD, and FASN in OMECs. These findings suggest that miR-199a-3p inhibited milk fat synthesis of OMECs. This is the first study to reveal the molecular mechanisms by which miR-199a-3p regulates the proliferation and milk fat synthesis of OMECs in sheep.

Modulation of endoplasmic reticulum stress via sulforaphane-mediated AMPK upregulation against nonalcoholic fatty liver disease in rats

Nonalcoholic fatty liver disease (NAFLD) is a major health concern. Endoplasmic reticulum (ER) stress, inflammation, and metabolic dysfunctions may be targeted to prevent the progress of nonalcoholic fatty liver disease. Sulforaphane (SFN), a sulfur-containing compound that is abundant in broccoli florets, seeds, and sprouts, has been reported to have beneficial effects on attenuating metabolic diseases. In light of this, the present study was designed to elucidate the mechanisms by which SFN ameliorated ER stress, inflammation, lipid metabolism, and insulin resistance — induced by a high-fat diet and ionizing radiation (IR) in rats. In our study, the rats were randomly divided into five groups: control, HFD, HFD + SFN, HFD + IR, and HFD + IR + SFN groups. After the last administration of SFN, liver and blood samples were taken. As a result, the lipid profile, liver enzymes, glucose, insulin, IL-1β, adipokines (leptin and resistin), and PI3K/AKT protein levels, as well as the mRNA gene expression of ER stress markers (IRE-1, sXBP-1, PERK, ATF4, and CHOP), fatty acid synthase (FAS), peroxisome proliferator–activated receptor-α (PPAR-α). Interestingly, SFN treatment modulated the levels of proinflammatory cytokine including IL-1β, metabolic indices (lipid profile, glucose, insulin, and adipokines), and ER stress markers in HFD and HFD + IR groups. SFN also increases the expression of PPAR-α and AMPK genes in the livers of HFD and HFD + IR groups. Meanwhile, the gene expression of FAS and CHOP was significantly attenuated in the SFN-treated groups. Our results clearly show that SFN inhibits liver toxicity induced by HFD and IR by ameliorating the ER stress events in the liver tissue through the upregulation of AMPK and PPAR-α accompanied by downregulation of FAS and CHOP gene expression.

Effect of genetic merit for backfat thickness and paternal breed on performance, carcass traits, and gene expression in subcutaneous adipose tissue of feedlot-finished steers

• Backfat thickness trait can be selected through genetic merit in Nellore and crossbred steers (½ Aberdeen Angus + ½ Nellore). • Animal performance is not affected by genetic merit for backfat thickness. • The expression of ACACA, ADIPOQ, FASN, PLIN 1, PPARG and SREBF1 genes in subcutaneous adipose tissue is not affected by genetic merit for backfat thickness. This study evaluated selection for backfat thickness based on genetic merit (GMB) in Nellore (N) and crossbred (½ Aberdeen Angus + ½ Nellore, AN) steers, as well as the effects of this selection on animal performance, carcass traits, and gene expression in subcutaneous adipose tissue (SAT). The animals were obtained by mating Nellore cows with Nellore or Aberdeen Angus bulls. Both paternal genetic groups were selected to have positive (+, high GMB) or negative (-, low GMB) expected progeny differences for backfat thickness. Seventy-six animals (14 N+, 20 N-, 20 AN+, and 22 AN-) were reared on pasture and finished in the feedlot. The animals were slaughtered, and SAT samples were collected to investigate the relative expression of the acetyl-CoA carboxylase alpha ( ACACA ), adiponectin, C1Q and collagen domain containing ( ADIPOQ ), fatty acid synthase ( FASN ), perilipin 1 ( PLIN1 ), peroxisome proliferator activated receptor gamma ( PPARG ), and sterol regulatory element binding protein ( SREBF1 ) genes. Carcass weight and yield, conformation, finishing score, fat distribution score, and measurements of backfat thickness and rib eye area were evaluated. There was no effect of GMB on performance or on the relative expression of the genes studied. High GMB steers had greater backfat thickness and higher finishing, conformation and fat distribution scores in the carcass than low GMB animals. The use of high GMB animals provides better carcass finishing without affecting performance. The greater fat deposition in carcasses of high GMB animals is not associated with the relative expression of the genes studied.

Juvenile Hormone Is an Important Factor in Regulating Aspongopus chinensis Dallas Diapause.

Aspongopus chinensis is a Chinese traditional edible and medicinal insect, which is in great demand in the society. This insect reproduces once a year which is caused by reproductive diapause resulting in insufficient production in wild resources. However, the mechanism of diapause in A. chinensis is still unclear. In this study, we focus on the relationship between juvenile hormones (JHs) and A. chinensis diapause. The results showed that JHIII concentration in diapause adult individuals was significantly lower than that in diapause termination adult individuals. When exogenous JHⅢ was injected into diapause adults, the rate of mating was increased significantly, development of the reproductive systems was accelerated, consumption of fat intensified, the expression of juvenile hormone acid o-methyl-transferase (JHAMT) was upregulated, and juvenile hormone epoxide hydrolase (JHEH) and fatty acid synthase (FAS) gene expressions were downregulated. In addition, RNAi of JHAMT decreased JH concentration, delayed the development of reproductive systems, slowed down fat consumption, and delayed the mean mating occurrence time significantly. Conversely, RNAi of JHEH resulted in an increased concentration of JH, development of reproductive systems was accelerated, consumption of fat was intensified, and mean mating occurrence time advanced significantly. Taken together, these findings uncovered that JH plays an important role in regulating reproductive diapause in A. chinensis and, thus, could provide a theoretical basis for further research on the diapause of A. chinensis.

Dietary protein/carbohydrate ratio and feeding frequency affect feed utilization, intermediary metabolism, and economic efficiency of gilthead seabream (Sparus aurata) juveniles

To evaluate the effects of dietary protein/carbohydrate (P/CHO) ratio and feeding frequency (FF) on growth, intermediary metabolism, and economic efficiency of gilthead seabream (Sparus aurata) juveniles, two practical isolipidic (17%) diets were formulated to include high protein (50%)/ low starch (10%) (diet P50/CHO10) or low protein (40%)/ high starch (20%) (diet P40/CHO20). Triplicate groups of fish with 9.1 ± 0.01 g were fed for 60 days with these diets until visual satiation at three FF: one (9:00), two (9:00 and 17:00), or three (9:00, 13:00, and 17:00) meals per day. Dietary P/CHO ratios did not affect growth performance while feeding 2 or 3 meals per day improved fish growth. Fish fed diet P40/CHO20 had increased feed intake (FI), protein efficiency ratio (PER), and nitrogen retention (NR), and lower feed efficiency (FE), nitrogen intake (NI), and economic conversion ratio (ECR). Feeding 2 or 3 meals per day increased FI, NI, ECR, and economic profit index, and decreased FE, PER, and NR. Fish fed diet P40/CHO20 presented increased hepatic lipid and glycogen content, hepatocyte area covered by lipid vacuoles, and glucokinase (gk) gene expression, and decreased glutamate dehydrogenase expression. Fish fed 3 meals per day had decreased plasma triglycerides and total protein levels, while fish fed 2 or 3 meals per day presented decreased hepatic growth hormone receptor-i (ghr-i), gk, and fatty acid synthase gene expression. Interaction between P/CHO ratio and FF was only observed in plasmatic glucose, cholesterol, and total lipids levels, and insulin-like growth factor-1, and ghr-ii gene expression. Overall, glycogenesis, glycolysis, and economic efficiency seemed to be increased while the amino acid catabolism was reduced in fish fed the P40/CHO20 diet. Higher FF increased growth and economic efficiency, and reduced glycolysis and lipogenesis pathways. In conclusion, a diet with P40/CHO20 ratio fed twice a day appears to be the most adequate strategy regarding feed utilization and economic efficiency for gilthead seabream juveniles in order to achieve optimum sustainable aquaculture.

Evaluation of the expression of fatty acid synthase and O-GlcNAc transferase in patients with liver cancer by exploration of transcriptome databases and experimental approaches.

Tumor occurrence and development are closely related to metabolism abnormalities. One of the metabolic networks that is dysregulated during carcinogenesis is the fatty acid synthesis pathway, which is mainly controlled by fatty acid synthase (FASN). We previously demonstrated in proliferating HepG2 liver cancer cells that FASN expression depends on the catalytic activity of O-GlcNAc transferase (OGT) and the activation of the mechanistic/mammalian target of rapamycin (mTOR) pathway. The aim of the present study was to go further in these investigations by analyzing datasets and tissues of patients with liver cancer. To that purpose, transcriptome databases were explored, and reverse transcription-quantitative PCR, western blotting and immunohistochemistry were used. Database analyses revealed that FASN and OGT gene expression was higher in certain cancer tissues, including liver hepatocellular carcinoma, compared with that in non-cancerous tissues. At the protein level, FASN expression was higher in the liver cancer-derived cell lines HepG2 and Hep3B compared with the immortalized human hepatocytes IHH cell line. However, neither the expression of OGT nor of its product O-GlcNAcylation showed any significant difference among the three hepatic cell lines. Subsequently, the expression of FASN and OGT at the protein and mRNA levels was evaluated in human liver cancer and non-tumoral tissues from the same patients with different liver lesions. The results from western blotting demonstrated a significant increase in OGT ands O-GlcNAcylation expression in liver cancer tissues independently of the type of lesion characterizing the non-tumoral counterpart. As previously reported for HepG2 proliferating cells, the protein level of FASN was positively correlated with the activation of mTOR and, although a rather upward trend, a high variability in its expression was monitored between patients. However, the results from immunohistochemistry showed no particular modification for OGT and O-GlcNAcylation expression and a significant increase in FASN expression in cancer tissues compared with that in adjacent non-tumoral tissues. Non-significant changes were observed for FASN and OGT mRNA levels between tumoral and non-tumoral samples, with a high variability between patients. Taken together, these results demonstrated that FASN expression was higher in hepatic cancer tissues in comparison with non-tumoral tissues. Furthermore, OGT expression and activity were shown to vary greatly between cell or cancer type, making any generalization difficult.

Association analysis between FASN genotype and milk traits in Mediterranean buffalo and its expression among different buffalo tissues

Fatty acid synthase (FASN) is a multifunctional protein that catalyzes the synthesis of long-chain saturated fatty acid. In this study, we identified the single nucleotide polymorphisms (SNPs), and their association with milk traits in Mediterranean buffalo, and the expression of FASN gene in different tissues was measured. Nine SNPs (g.-1640G > A, g.-1099C > T, g.1095C > A, g.3221G > A, g.4762G > A, g.5299G > A, g.7164G > A, g.7272T > C, and g.8927T > C) were identified by DNA pooled sequencing and then genotyped. Seven identified SNPs except g.3221G > A and g.8927T > C were found significantly associated with both fat and protein percentage, and also the g.7164G > A and g.8927T > C had significant association with peak milk yield and protein percentage, respectively. One haplotype block was successfully constructed by linkage disequilibrium (LD) analysis and it showed a significant association with both fat percentage and protein percentage. Expression of FASN gene was found in almost all the buffalo tissues including mammary gland, heart, liver, spleen, lung, kidney, uterus, and ovary, and to be highest in lung and mammary gland. Our findings suggest that polymorphisms in the buffalo FASN gene are associated with milk production traits and can be used as a candidate gene for milk traits and marker-assisted selection in buffalo breeding program.

Effects of uni and bilateral castration on growth performance and lipid metabolism in yellow cattle

This study was conducted to examine the influence of uni and bilateral castration on growth performance and lipid metabolism in yellow cattle. Eighteen 9-month-old healthy yellow cattle (average body weight 184.03 ± 4.09 kg) were selected and divided into three groups: The uncastrated cattle (C), half castrated cattle (HC) and full castrated cattle (FC). The results showed that the growth rate of FC group was significantly reduced as compared to HC and C group, while the feed to gain ratio exhibited an opposite trend. The concentrations of triglycerides (TG), low-density lipoprotein cholesterol (LDL) and high-density lipoprotein (HDL) were increased significantly in FC group from day 60 to the end of the trial compared to HC and control groups. Serum testosterone concentration of FC group cattle was decreased from day 60 to 120 d of the trial compared to HC and control groups. The concentration of the lauric acid in FC cattle was significantly increased from the HC and control groups. In the FC group, the acetyl-CoA carboxylase alpha (ACACA), ACC and fatty acid synthase (FAS) gene expression levels were significantly higher compared to control and HC groups. Our results of this study suggest that bilateral castration increased the lipid metabolism and fatty acid composition compared to unilateral castrated and un-castrated cattle. Highlights Bilateral castration alters the growth performance in yellow cattle. Bilateral castration alters hormones levels and lipid metabolites levels in serum. Bilateral castration improves the lipid metabolism and fatty acid profile.

Effects of dietary starch levels on growth, feed utilization, glucose and lipid metabolism in non-transgenic and transgenic juvenile common carp (Cyprinus carpio L.)

This experiment investigated the effects of dietary starch on growth performance, feed utilization, glucose and lipid metabolism of non-transgenic and growth hormone (GH) transgenic juvenile common carp (Cyprinus carpio L.). Transgenic common carp (TG fish) and non-transgenic (NT fish) were fed diets with five starch levels (0, 10%, 20%, 30%, 40%). After 41 days, TG fish showed higher growth and feed utilization at low-starch diets, while similar growth, lower feeding rate (FR) and slightly higher feed efficiency (FE) was observed at high-starch diets. Lower plasma glucose level and hepatic glycogen content were observed in TG fish, which could be related to the higher glycolysis (high gene expression of hexokinase (hk), p &lt; 0.01) and gluconeogenesis (high gene expression of fructose-1,6-bisphosphatase (fbpase), p &lt; 0.01; glucose-6-phosphatase (g6pase), p &lt; 0.01) of TG fish. Compared to NT fish, weakened fat synthesis (lower gene expression of fatty-acid synthase (fas), p &lt;0.01; acetyl-CoA carboxylase 1 (acc1), p &lt; 0.01; acyl-CoA desaturase 1 (scd1), p = 0.011) and enhanced fatty acid oxidation (high gene expression of hormone-sensitive lipase (hsl), p =0.01; carnitine palmitoyl transferase 1a (cpt-1a), p &lt; 0.01), led to the decrease of body lipid content in TG fish. On the other hand, with increased dietary starch, increased body lipid and hepatic glycogen contents were observed in both TG and NT fish, suggesting that high dietary starch levels promoted glycolysis, fat synthesis and inhibited gluconeogenesis, fatty acid β-oxidation. Overall, TG fish showed higher growth performance at low starch diets, and higher ability of glycolysis, fatty acid oxidation and lower lipid synthesis than the NT fish.

Preliminary study on effect of Gegen Qinlian Decoction on enzyme activity, gene expression and methylation level of FASN in adipose tissue of rats with insulin resistance

To investigate the effect of Gegen Qinlian Decoction(GQD) on enzyme activity, gene expression and methylation level of fatty acid synthase(FASN) in adipose tissue from rats with insulin resistance induced by high-fat diet. The 60% fat-powered high-fat diet was continuously given to male SD rats to induce the insulin resistance model. Then, they were divided into five groups randomly and administrated by gavage every day for 16 weeks with following drugs respectively: 10 mL·kg~(-1)water for control group(C) and insulin resistance model control group(IR), 1.65 g·kg~(-1)GQD per day for low-dose group(GQDL), 4.95 g·kg~(-1)GQD per day for medium-dose group(GQDM), 14.85 g·kg~(-1)GQD per day for high-dose group(GQDH), and 5 mg·kg~(-1) rosiglitazone per day for rosiglitazone group(RGN). Epididymal adipose tissue was taken to determine enzyme activity of FASN by colorimetric method, mRNA expression level of Fasn by quantitative Real-time PCR(Q-PCR) and CpGs methylation level between +313 and +582 by bisulfite sequencing PCR(BSP). These results showed that Fasn expression was significantly lowered in IR model rats compared with the control rats(P&lt;0.01). Enzymatic activity and CpGs methylation level of Fasn in IR group showed downward trends. Low and medium-dose GQD can increase enzyme activity of FASN(P&lt;0.05). Moreover, low-dose GQD increased the total CpGs methylation level of Fasn fragment between +313 and +582 in insulin resistance rats(P&lt;0.05). For GQDM group, the methylation frequency of CpGs at positions +506 and +508(P&lt;0.01) as well as the methylation frequency of CpGs on the binding sites of transcription factorzinc finger protein 161(P&lt;0.05) were significantly increased. The methylation frequency of CpG at +442 position was positively correlated with Fasn expression(P&lt;0.01, r=0.735), and methylation frequencies of CpGs at +345 and +366 positions were positively associated to enzyme activity of FASN respectively(P&lt;0.05, r=0.479; P&lt;0.01, r=0.640). In conclusion, GQD can reverse enzyme activity of FASN and methylation level of Fasn in adipose tissue of insulin resistant rats, and CpG sites at positions +506 and +508 may be the targets of GQD. The methylation level of CpGs at + 345 and + 366 sites were possibly related to FASN activity, while methylation of CpG at + 442 site may be closely correlated with mRNA level of Fasn. In addition, GQD did not significantly change mRNA expression level of Fasn, but effectively reversed enzymatic activity, suggesting that GQD may regulate the post transcriptional expression of Fasn.

Progesterone receptor isoform-dependent cross-talk between prolactin and fatty acid synthase in breast cancer.

Progesterone receptor (PR) isoforms can drive unique phenotypes in luminal breast cancer (BC). Here, we hypothesized that PR-B and PR-A isoforms differentially modify the cross-talk between prolactin and fatty acid synthase (FASN) in BC. We profiled the responsiveness of the FASN gene promoter to prolactin in T47Dco BC cells constitutively expressing PR-A and PR-B, in the PR-null variant T47D-Y cell line, and in PR-null T47D-Y cells engineered to stably re-express PR-A (T47D-YA) or PR-B (T47D-YB). The capacity of prolactin to up-regulate FASN gene promoter activity in T47Dco cells was lost in T47D-Y and TD47-YA cells. Constitutively up-regulated FASN gene expression in T47-YB cells and its further stimulation by prolactin were both suppressed by the prolactin receptor antagonist hPRL-G129R. The ability of the FASN inhibitor C75 to decrease prolactin secretion was more conspicuous in T47-YB cells. In T47D-Y cells, which secreted notably less prolactin and downregulated prolactin receptor expression relative to T47Dco cells, FASN blockade resulted in an augmented secretion of prolactin and up-regulation of prolactin receptor expression. Our data reveal unforeseen PR-B isoform-specific regulatory actions in the cross-talk between prolactin and FASN signaling in BC. These findings might provide new PR-B/FASN-centered predictive and therapeutic modalities in luminal intrinsic BC subtypes.

Regulatory effect of diosgenin on lipogenic genes expression in high-fat diet-induced obesity in mice.

Obesity is one of the most serious health problems in the world, increasing the risk of other chronic diseases. Alterations in fatty acid synthesis related genes are crucially involved in obesity progression. Diosgenin (DG) was one of the phytosterols compounds with vital activity against lipid disorders. Therefore, this study was intended to evaluate the protective effect of DG on lipogenesis in the high-fat diet (HFD)-induced obesity in mice, via investigating the expression of two of the fatty acid synthesis–involved genes; sterol regulatory element-binding protein (SREBP-1c) and fatty acid synthase (FASN) genes. Thirty adult male mice were divided into 3 groups. Control group, fed with normal diet; HFD group, mice fed with a high-fat diet and HFD + DG group, mice fed with a high-fat diet and supplemented in parallel with DG for 6 consecutive weeks. The effect of DG on Body weights, liver enzymes, lipid profile, were evaluated. Histopathological fatty changes as well as SREBP-1c and FASN gene expression were also investigated. DG significantly alleviated body weight gain, adjusted liver enzymes, and improved lipid profile. Additionally, DG ameliorated the histopathological changes by reducing the lipid vacuoles and hence the hepatosteatosis. Accordingly, DG significantly downregulated the two-fold increase in the SREBP-1c and FASN gene expression observed in the HFD group. In conclusion, DG possesses a beneficial impact against diet-induced obesity in mice, which makes it a good candidate for NAFLD and obesity prevention.

Chlorogenic Acid Decreases Lipid Accumulation in 3T3-L1 Adipocytes by Modulating the Transcription Factors

The aim of this study is to investigate the effect of chlorogenic acid (CA) on 3T3-L1 adipocytes and its mechanism action. CA at 1-25 μM showed no cytotoxicity to 3T3-L1 pre-adipocytes and 3T3-L1 adipocytes. CA significantly inhibited oil red O-stained material (OROSM) and intracellular triglyceride levels in 3T3-L1 adipocytes in a concentration-dependent manner. In addition, CA down-regulated the glycerol-3–phosphate dehydrogenase (GPDH) and peroxisome proliferator–activated receptor γ (PPARγ) activity. A real-time polymerase chain reaction revealed that CA inhibited the PPARγ, CCAAT/enhancer-binding protein alpha (C/EBPα), sterol regulatory element binding protein-1c (SREBP-1c), and fatty acid synthase (FAS) gene expression, which may in part account for anti-adipogenesis of CA. Thus, CA could act as a potential lipid lowering functional resource.

Transcriptional analyses of the effects of Catharanthus roseus L. medicinal plant extracts on some markers related to obesity and inflammation in 3T3-L1 mouse cell lines

Catharanthus roseus L. (C. roseus) is one of the medicinal plants used to treat diabetes. In this study, 3T3-L1 preadipocyte cell lines which are fully differentiated into adipocytes were utilized and these were treated with extracts derived from above ground part of C. roseus. The effect of these extracts on obesity and inflammation markers in cells was examined at the transcriptional level. Adipocyte lipid contents were measured by Oil Red O staining. Analyses, including changes related to adiposity and inflammation, were evaluated by measuring the relative mRNA expression levels of the genes of interest by the Real Time Reverse Transcription-Polymerase Chain Reaction (RT-PCR) method. The appropriate dose and durations for C. roseus were determined to be 12.5 μg/mL and 24- and 48 h respectively. The expression of the inflammation marker Interleukin-6 (Il-6) was decreased when C. roseus extract was administered to fully differentiated 3T3-L1 cells according to the determined dose and durations. Lipoprotein lipase (Lpl) and Fatty acid synthase (Fasn) gene expressions in fully differentiated cells decreased compared to the control after 24 h however, this effect was not observed after 48 h. Collagen V, has also been shown to be affected by treatment of fully differentiated 3T3-L1 cells with plant extracts in both 24- and 48-h periods.