Fatty Acid Synthase Protein Research Articles

Insight View on the Role of in Ovo Feeding of Clenbuterol on Hatched Chicks: Hatchability, Growth Efficiency, Serum Metabolic Profile, Muscle, and Lipid-Related Markers.

Simple SummaryThis study examined the effects of ovo injection of clenbuterol on fat deposition and growth performance in chickens, which is prejudicial to poultry consumers and muscle growth-related genes, egg hatchability, and fertility. The achieved result showed a definite effect of clenbuterol on body gain and hatchability. It decreased fat deposition and upregulation of muscle growth-related gene expressions accompanied by modulation of fatty and amino acid composition, reflecting a new insight into the intracellular pathways of clenbuterol supplementation on chicks.The present study aimed to assess the in ovo administration of clenbuterol on chick fertility, growth performance, muscle growth, myogenic gene expression, fatty acid, amino acid profile, intestinal morphology, and hepatic lipid-related gene expressions. In this study, 750 healthy fertile eggs from the local chicken breed Dokki-4 strain were analyzed. Fertile eggs were randomly divided into five experimental groups (150 eggs/3 replicates for each group). On day 14 of incubation, in addition to the control group, four other groups were established where 0.5 mL of worm saline (30 °C) was injected into the second group of eggs. In the third, fourth, and fifth groups, 0.5 mL of worm saline (30 °C), 0.9% of NaCl, and 10, 15, and 20 ppm of clenbuterol were injected into the eggs. Results suggested that clenbuterol increased growth efficiency up to 12 weeks of age, especially at 15 ppm, followed by 10 ppm, decreased abdominal body fat mass, and improved hatchability (p < 0.01). Clenbuterol also modulated saturated fatty acid levels in the breast muscles and improved essential amino acids when administered at 10 and 15 ppm. Additionally, clenbuterol at 15 ppm significantly decreased myostatin gene expression (p < 0.01) and considerably increased IGF1r and IGF-binding protein (IGFBP) expression. Clenbuterol administration led to a significant upregulation of hepatic PPARα, growth hormone receptor, and Lipoprotein lipase (LPL) mRNA expression with a marked decrease in fatty acid synthase (FAS) and sterol regulatory element-binding protein 1 (SREBP-1c) expression. In conclusion, the current study revealed that in ovo injection of clenbuterol showed positive effects on the growth of hatched chicks through reduced abdominal fat deposition, improved intestinal morphology, and modulation of hepatic gene expressions in myogenesis, lipogenesis, and lipolysis.

Daily Dose of Bovine Lactoferrin Prevents Ethanol-Induced Liver Injury and Death in Male Mice by Regulating Hepatic Alcohol Metabolism and Modulating Gut Microbiota.

Lactoferrin (Lf) possess a protective potential to liver, but whether it can prevent alcoholic liver injury (ALI) remains unclear. Four groups of male C57BL/6J mice are fed with different diets, namely, AIN-93G diet for control (CON) and ethanol (EtOH) groups, and AIN-93G diet with 0.4% and 4% casein replaced by Lf for low-dose Lf (LLf) and high-dose Lf (HLf) groups, respectively. ALI is induced by giving 20% ethanol ad libitum combined with four "binges". Lf can remarkably decrease EtOH-induced mortality. Lf promotes aldehyde dehydrogenase-2 (ALDH2) expression and suppressing cytochrome P450 2E1 (CYP2E1) overexpression, resulting in the reduced hepatic superoxide and inflammation levels, which ultimately leads to the hepatic injury alleviation. However, HLf increases acetyl-CoA carboxylase and fatty acid synthase protein levels, which suggests that excessive intake may weaken the beneficial effects of Lf. Moreover, LLf increases the relative abundances of Akkermansia and Lactobacillus. Additionally, the study shows that Lf likely exerts action in its digestive product forms rather than intact Lf molecular in normal condition. LLf can ameliorate ALI, which is associated with the regulation of hepatic alcohol metabolism and the modulation of gut microbiota. However, excessive Lf intake may result in a diminished benefit.

Excessive Dietary Lipid Affecting Growth Performance, Feed Utilization, Lipid Deposition, and Hepatopancreas Lipometabolism of Large-Sized Common Carp (Cyprinus carpio).

An 82-day study was conducted to assess the effect of the dietary lipid levels on growth performance, feed utilization, lipid deposition, and hepatopancreas lipometabolism of large-sized common carp (Cyprinus carpio). Six isonitrogenous (300 g/kg protein) pelletized diets with different dietary lipid levels (30, 60, 90, 120, 150, and 180 g/kg) were fed in triplicate to fish groups with 75 individuals (with an initial mean weight of 247.00 ± 16.67 g). The results showed that there was a significant increase in weight gain (WG) rate (WGR), specific growth rate (SGR), and protein efficiency ratio (PER) as dietary lipid levels increased from 30 to 60 g/kg (p < 0.05) and then there was a decline. Feed conversion rate (FCR) was observed to be significantly lower in 60 g/kg lipid treatments (p < 0.05). Muscle crude protein contents were obtained to be significantly higher in 60 and 90 g/kg treatments (p < 0.05). The crude lipid content in the hepatopancreas increased significantly with an increase in dietary lipid levels (p < 0.05). The expression of lipoprotein lipase (LPL) and carnitine palmitoyltransferase-1 (CPT1) in the hepatopancreas was significantly downregulated with an increase in dietary lipid levels while the expression of growth hormone (GH), insulin-like growth factor-1 (IGF-1), fatty acid synthase (FAS), acetyl-CoA carboxylase-1 (ACC-1), and sterol regulatory element binding protein (SREBP) was upregulated first in 30 and 60 g/kg lipid treatments and then downregulated significantly in other treatments. The results revealed that excess dietary lipid supplements (more than 60 g/kg) would inhibit WG and would aggravate the lipid decomposition in the hepatopancreas. Based on WGR and FCR, the dietary lipid levels of 59.5 and 70.4 g/kg were optimal for growth performance and feed utilization of large-sized common carp.

Triclosan down-regulates fatty acid synthase through microRNAs in HepG2 cells

Triclosan is a promising candidate of fatty acid synthase (FASN) inhibitor by blocking FASN activity, but its effect on FASN expression and the underling epigenetic mechanism remain elusive. In this study, the effect of triclosan on FASN mRNA and protein expressions in human HepG2 cells and the regulatory role of microRNAs (miRNAs) in the downregulation of FASN induced by triclosan were explored through experiments and bioinformatics analysis. The results showed that triclosan not only directly inhibited FASN activity, but also significantly decreased FASN mRNA and protein levels in human liver HepG2 cells. Nine miRNAs targeting FASN mRNA degradation were identified by miRNA prediction tools, and the expression levels of these nine miRNAs were then detected by real-time quantitative PCR. Triclosan significantly increased the expressions of the six miRNAs, namely miR-15a, miR-107, miR-195, miR-424, miR-497 and miR-503, leading to the downregulation of FASN. Further investigation revealed that the six triclosan-upregulated miRNAs played an important regulatory role in lipid metabolism and cell cycle by gene ontology annotations and pathway analysis. Consistent with the results of bioinformatics analyses, triclosan significantly reduced the intracellular lipid content by triglyceride assay, oil red O, BODIPY 493/503 and Nile Red staining, thereby inhibiting the growth of HepG2 cells through apoptosis. Taken together, our study reveals that triclosan downregulates FASN expression through a variety of miRNAs, providing new insight for triclosan as a FASN inhibitor candidate to regulate lipid metabolism in human hepatoma cells.

Dose-Effect of a 6-week treatment with PEP2DIA®, a Patented Milk Protein Hydrolysate on Sucrose Tolerance in Goto-Kakizaki (GK) Rats

Abstract Objectives The study was designed to evaluate the dose-effect of PEP2DIA(r)(PEP), a milk protein hydrolysate on glycemic control of type 2 diabetic Goto-Kakizaki (GK) rats treated for 6 weeks. Methods Rats (n = 10) were treated with PEP solubilized in spring water by oral gavage once a day for 6 weeks. The effects of a 6-week-administration of PEP (63 mg/kg, 88.6 mg/kg and 126 mg/kg per os) were evaluated on sucrose tolerance and insulin response to sucrose in type 2 diabetic GK rats from weaning (3 weeks) to adult age (9 weeks). Circulating GLP-1 concentration and DDP-4 activity, α-glucosidase activity in duodenum and jejunum, Fatty Acid Synthase (FAS) and Sterol regulatory element-binding protein 1 (SREBP-1c) gene expressions in liver and retroperitoneal adipose tissue were also measured. PEP is a patented milk protein hydrolysate containing AP dipeptides which inhibit alpha glucosidase. The alpha-glucosidase inhibitor Acarbose (40 mg/kg) was used as reference. Results The 6 week-treatment with PEP at 3 doses improved sucrose tolerance with the best effect at the dose of 63 mg/kg. Insulin response to sucrose was lower than control after PEP treatment at all the doses tested with the strongest decrease with 63 mg/kg of PEP. This decrease in insulin response seems to be at least in part the consequence of an improvement of the insulin resistance of the GK rats. At the lowest dose tested (63 mg/kg), FAS and SREBP-1c gene expressions were significantly decreased in retroperitoneal adipose tissue of GK rats, suggesting that PEP inhibited lipogenesis. PEP treatment induced strong increases in GLP-1 plasma level at all the doses tested but the difference reached significance only with 63 and 126 mg/kg of PEP. An inhibition of alpha-glucosidase in duodenum but not in jejunum was observed after the 6-week-treatment with PEP. Moreover, in retroperitoneal adipose tissue, PEP at the lowest dose tested (63 mg/kg), significantly decreased gene expression of both SREBP-1c and FAS, suggesting a beneficial effect on triglyceride accumulation in adipose tissue Conclusions These results showed that PEP2DIA(r) could have a beneficial effect on glycemic control of type 2 diabetic GK rats and suggest that this milk protein hydrolysate should be beneficial Impaired Fasting Glucose or Impaired Glucose Tolerance subjects for delaying the progression to overt type 2 diabetes. Funding Sources INGREDIA S.A.

Modest sleep restriction does not influence steps, physical activity intensity or glucose tolerance in obese adults.

Sleep restriction (SR) (<6h) and physical activity (PA) are risk factors for obesity, but little work has examined the inter-related influences of both risk factors. In a free-living environment, 13 overweight/obese adults were sleep restricted for five nights to 6h time-in-bed each night, with and without regular exercise (45min/65% VO2 max; counterbalanced design). Two days of recovery sleep followed SR. Subjects were measured during a mixed meal tolerance test (MMT), resting metabolic rate, cognitive testing and fat biopsy (n=8). SR increased peak glucose response (+7.3mg/dl, p=.04), elevated fasting non-esterified fatty acid (NEFA) concentrations (+0.1mmol/L, p=.001) and enhanced fat oxidation (p<.001) without modifying step counts or PA intensity. Inclusion of daily exercise increased step count (+4,700 steps/day, p<.001) and decreased the insulin response to a meal (p=.01) but did not prevent the increased peak glucose response or elevated NEFA levels. The weekend recovery period improved fasting glucose (p=.02), insulin (p=.02), NEFA concentrations (p=.001) and HOMA-IR (p<.01) despite reduced steps (p<.01) and increased sedentary time (p<.01). Abdominal adipose tissue (AT) samples, obtained after baseline, SR and exercise, did not differ in lipolytic capacity following SR. Fatty acid synthase protein content tended to increase following SR (p=.07), but not following exercise. In a free-living setting, SR adversely affected circulating NEFAs, fuel oxidation and peak glucose response but did not directly affect glucose tolerance or AT lipolysis. SR-associated metabolic impairments were not mitigated by exercise, yet recovery sleep completely rescued its adverse effects on glucose metabolism.

Carnosic Acid and Carnosol Activate AMPK, Suppress Expressions of Gluconeogenic and Lipogenic Genes, and Inhibit Proliferation of HepG2 Cells.

Carnosic acid (CA), carnosol (CL) and rosmarinic acid (RA), components of the herb rosemary, reportedly exert favorable metabolic actions. This study showed that both CA and CL, but not RA, induce significant phosphorylation of AMP-dependent kinase (AMPK) and its downstream acetyl-CoA carboxylase 1 (ACC1) in HepG2 hepatoma cells. Glucose-6-phosphatase (G6PC) and phosphoenolpyruvate carboxykinase 1 (PCK1), rate-limiting enzymes of hepatic gluconeogenesis, are upregulated by forskolin stimulation, and this upregulation was suppressed when incubated with CA or CL. Similarly, a forskolin-induced increase in CRE transcriptional activity involved in G6PC and PCK1 regulations was also stymied when incubated with CA or CL. In addition, mRNA levels of ACC1, fatty acid synthase (FAS) and sterol regulatory element-binding protein 1c (SREBP-1c) were significantly reduced when incubated with CA or CL. Finally, it was shown that CA and CL suppressed cell proliferation and reduced cell viability, possibly as a result of AMPK activation. These findings raise the possibility that CA and CL exert a protective effect against diabetes and fatty liver disease, as well as subsequent cases of hepatoma.

The evaluation for CpG island methylation of FASN and SREBP promoter in Mongolian gerbils NAFLD model

IntroductionThe methylation of CpG island in promoter and its nearby area is one of the most important way to inhibit gene expression. The aim of this study was to examine the potential relationship between the CpG island methylation for fatty acid synthase (FASN) and sterol regulatory element-binding protein 1 (SREBP-1).Material and methodsTen newborn pups of gerbils as NB group, 10 adult gerbils with normal diet as control group, 10 adults with high-fat diet as NAFLD group, and 10 8-month-old as aged group. Blood and liver samples were collected for serum lipid detection and histopathology. A pyrosequencing technique was employed to determine the methylation rate. Then, the transcription and expression level for FASN and SREBP-1 were verified.ResultsSerum chelosterol and triglyceride was significantly increased in NAFLD and aged group (vs. control, P&lt;0.05). The gerbils in NAFLD and aged group also showed obvious hepatic steatosis confirmed by histological examination. Control group had the highest methylation rate for FASN and SREBP-1, which were reduced in NAFLD and aged group. Except for NB group, both the transcription and expression levels of SREBP-1 and FASN genes were presented control group&gt; Aged group &gt; NAFLD group. Genes of SREBP and FASN showed a trend of hypomethylation in the NAFLD gerbil model.ConclusionsThe expression of SREBP gene tended to decrease, while the expression of FASN gene tended to increase with the age and disease development. FASN and SREBP-1 methylation might be a new method to evaluate NAFLD animal model and an available target for genetic marker screening.

Heat stress promotes lipid accumulation by inhibiting the AMPK-PGC-1α signaling pathway in 3T3-L1 preadipocytes.

Heat stress (HS) results in health problems in animals. This study was conducted to investigate the effect and the underlying mechanism of HS on the proliferation and differentiation process of 3T3-L1 preadipocytes. 3T3-L1 preadipocytes were treated at 37 °C or 41.5 °C. HS up-regulated the mRNA and protein expression level of heat shock protein 70 (HSP70). Furthermore, the proliferation of 3T3-L1 preadipocytes were significantly inhibited after HS treatment for 2 days. A large number of accumulated lipid droplets were observed under the microscope after HS treatment for 8 days. Notably, the result of oil red O staining showed that the number of lipid droplets increased significantly and the differentiation ability of the cells was enhanced after HS. Moreover, after 2 and 8 d of differentiation, HS increased the transcription levels of fat synthesis genes including peroxisome proliferators activated receptor γ (PPARγ), fatty acid binding protein 2 (AP2), fatty acid synthase (FAS) and CCAAT enhancer binding protein α (CEBPα) genes, while decreasing the transcription levels of lipid decomposition genes including ATGL and HSL genes. In addition, HS reduced the expression of AMPK and PGC-1α, as well as the dephosphorylation of AMPK. 5-Aminoimidazole-4-carboxamide ribonucleotide (AICAR) can eliminate HS induced lipogenesis by activating AMPK. These results indicated that HS inhibited the proliferation of 3T3-L1 preadipocytes and promoted lipid accumulation by inhibiting the AMPK-PGC-1α signaling pathway in 3T3-L1 preadipocytes. This work lays a theoretical foundation for improving the effect of HS on meat quality of livestock and provides a new direction for the prevention of obesity caused by HS.

Fatty Acid Synthase Confers Tamoxifen Resistance to ER+/HER2+ Breast Cancer.

Simple SummaryOveractivation of the human epidermal growth factor receptor 2 (HER2) is one of the main drivers of tamoxifen resistance in estrogen receptor (ER)-positive breast cancer patients. Combined targeting of HER2 and ER, however, has yielded disappointing results in the clinical setting. Therefore, other potential mechanisms for tamoxifen resistance would not be overcome by solely blocking the cross-talk between ER and HER2 at the receptor(s) level. Using cell lines, animal models, and clinical data, we provide evidence to support a critical role of fatty acid synthase (FASN)—the major site for endogenous fat synthesis—in HER2-driven tamoxifen resistance. Importantly, treatment with a FASN inhibitor impeded the estrogen-like tumor-promoting effects of tamoxifen and fully restored the anti-estrogenic activity of tamoxifen in ER+/HER2-overexpressing breast cancer xenografts. We postulate FASN as a biological determinant of HER2-driven tamoxifen resistance and FASN inhibition as a novel therapeutic approach to restore tamoxifen sensitivity in endocrine-resistant breast cancer.The identification of clinically important molecular mechanisms driving endocrine resistance is a priority in estrogen receptor-positive (ER+) breast cancer. Although both genomic and non-genomic cross-talk between the ER and growth factor receptors such as human epidermal growth factor receptor 2 (HER2) has frequently been associated with both experimental and clinical endocrine therapy resistance, combined targeting of ER and HER2 has failed to improve overall survival in endocrine non-responsive disease. Herein, we questioned the role of fatty acid synthase (FASN), a lipogenic enzyme linked to HER2-driven breast cancer aggressiveness, in the development and maintenance of hormone-independent growth and resistance to anti-estrogens in ER/HER2-positive (ER+/HER2+) breast cancer. The stimulatory effects of estradiol on FASN gene promoter activity and protein expression were blunted by anti-estrogens in endocrine-responsive breast cancer cells. Conversely, an AKT/MAPK-related constitutive hyperactivation of FASN gene promoter activity was unaltered in response to estradiol in non-endocrine responsive ER+/HER2+ breast cancer cells, and could be further enhanced by tamoxifen. Pharmacological blockade with structurally and mechanistically unrelated FASN inhibitors fully impeded the strong stimulatory activity of tamoxifen on the soft-agar colony forming capacity—an in vitro metric of tumorigenicity—of ER+/HER2+ breast cancer cells. In vivo treatment with a FASN inhibitor completely prevented the agonistic tumor-promoting activity of tamoxifen and fully restored its estrogen antagonist properties against ER/HER2-positive xenograft tumors in mice. Functional cancer proteomic data from The Cancer Proteome Atlas (TCPA) revealed that the ER+/HER2+ subtype was the highest FASN protein expressor compared to basal-like, HER2-enriched, and ER+/HER2-negative breast cancer groups. FASN is a biological determinant of HER2-driven endocrine resistance in ER+ breast cancer. Next-generation, clinical-grade FASN inhibitors may be therapeutically relevant to countering resistance to tamoxifen in FASN-overexpressing ER+/HER2+ breast carcinomas.

Desulfovibrio vulgaris, a potent acetic acid-producing bacterium, attenuates nonalcoholic fatty liver disease in mice

ABSTRACT The emerging evidence supports the use of prebiotics like herb-derived polysaccharides for treating nonalcoholic fatty liver disease (NAFLD) by modulating gut microbiome. The present study was initiated on the microbiota-dependent anti-NAFLD effect of Astragalus polysaccharides (APS) extracted from Astragalus mongholicus Bunge in high-fat diet (HFD)-fed mice. However, the exact mechanisms underlying the beneficial effects of APS on NAFLD formation remain poorly understood. Co-housing experiment was used to assess the microbiota dependent anti-NAFLD effect of APS. Then, targeted metabolomics and metagenomics were adopted for determining short-chain fatty acids (SCFAs) and bacteria that were specifically enriched by APS. Further in vitro experiment was carried out to test the capacity of SCFAs-producing of identified bacterium. Finally, the anti-NAFLD efficacy of identified bacterium was tested in HFD-fed mice. Our results first demonstrated the anti-NAFLD effect of APS in HFD-fed mice and the contribution of gut microbiota. Moreover, our results indicated that SCFAs, predominantly acetic acid were elevated in APS-supplemented mice and ex vivo experiment. Metagenomics revealed that D. vulgaris from Desulfovibrio genus was not only enriched by APS, but also a potent generator of acetic acid, which showed significant anti-NAFLD effects in HFD-fed mice. In addition, D. vulgaris modulated the hepatic gene expression pattern of lipids metabolism, particularly suppressed hepatic fatty acid synthase (FASN) and CD36 protein expression. Our results demonstrate that APS enriched D. vulgaris is effective on attenuating hepatic steatosis possibly through producing acetic acid, and modulation on hepatic lipids metabolism in mice. Further studies are warranted to explore the long-term impacts of D. vulgaris on host metabolism and the underlying mechanism.

Neddylation of PTEN regulates its nuclear import and promotes tumor development.

PTEN tumor suppressor opposes the PI3K/Akt signaling pathway in the cytoplasm and maintains chromosomal integrity in the nucleus. Nucleus-cytoplasm shuttling of PTEN is regulated by ubiquitylation, SUMOylation and phosphorylation, and nuclear PTEN has been proposed to exhibit tumor-suppressive functions. Here we show that PTEN is conjugated by Nedd8 under high glucose conditions, which induces PTEN nuclear import without effects on PTEN stability. PTEN neddylation is promoted by the XIAP ligase and removed by the NEDP1 deneddylase. We identify Lys197 and Lys402 as major neddylation sites on PTEN. Neddylated PTEN accumulates predominantly in the nucleus and promotes rather than suppresses cell proliferation and metabolism. The nuclear neddylated PTEN dephosphorylates the fatty acid synthase (FASN) protein, inhibits the TRIM21-mediated ubiquitylation and degradation of FASN, and then promotes de novo fatty acid synthesis. In human breast cancer tissues, neddylated PTEN correlates with tumor progression and poor prognosis. Therefore, we demonstrate a previously unidentified pool of nuclear PTEN in the Nedd8-conjugated form and an unexpected tumor-promoting role of neddylated PTEN.

Mechanism of maggot debridement therapy in promoting wound angiogenesis in patients with diabetic foot ulcer

Mechanism of maggot debridement therapy in promoting wound angiogenesis in patients with diabetic foot ulcer

Transcriptomic analysis suggested the involvement of impaired lipid droplet biogenesis in graphene oxide-induced cytotoxicity in human umbilical vein endothelial cells

Previous studies revealed that direct contact with graphene oxide (GO) induced cytotoxic effects, but the importance of involvement of metabolic pathways, in particular lipid metabolism pathways, might be overlooked. In this study, human umbilical vein endothelial cells (HUVECs) were exposed to GO with large size (denoted as GO-L) or small size (denoted as GO-S), and transcriptomics were used to understand the mechanisms of cytotoxicity of GO at systemic levels. It was shown that GO-L more significantly induced cytotoxicity compared with GO-S. Transcriptomic analysis revealed that compared with GO-S, GO-L had larger impact on gene ontology terms related with mitochondrial function as well as Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways related with cell death and growth. But GO-S showed greater influence on KEGG pathways related with lipid metabolism. Both types of GO showed minimal impact on oxidative stress but increased de novo lipogenesis protein fatty acid synthase (FASN). However, only GO-S significantly promoted acyl-CoA synthetase 3 (ACSL3), a key enzyme responsible for esterification of free fatty acids and lipid droplet biogenesis. Not surprisingly, GO-L but not GO-S impaired lipid droplet biogenesis, and increasing lipid levels by oleic acid or α-linolenic acid reduced the cytotoxicity of GO-L to HUVECs. Combined, the results from this study suggested that impaired lipid droplet biogenesis was involved in GO-induced cytotoxicity in HUVECs, and inducing lipid droplet biogenesis could prevent the cytotoxicity of GO.

Orexin system is expressed in avian liver and regulates hepatic lipogenesis via ERK1/2 activation

Orexins are originally characterized as orexigenic hypothalamic neuropeptides in mammals. Subsequent studies found orexin to be expressed and perform pleiotropic functions in multiple tissues in mammals. In avian (non-mammalian) species, however, orexin seemed to not affect feeding behavior and its physiological roles are poorly understood. Here, we provide evidence that orexin and its related receptors are expressed in chicken hepatocytes. Double immunofluorescence staining showed that orexin is localized in the ER, Golgi, and in the lysosomes in LMH cells. Brefeldin A treatment reduced orexin levels in the culture media, but increased it in the cell lysates. Administration of recombinant orexins upregulated the expression of orexin system in the liver of 9-day old chicks, but did not affect feed intake. Recombinant orexins increased fatty acid synthase (FASN) protein levels in chicken liver, activated acetyl-CoA carboxylase (ACCα), and increased FASN, ATP citrate lyase(ACLY), and malic enzyme (ME) protein expression in LMH cells. Blockade ERK1/2 activation by PD98059 attenuated these stimulating effects of orexin on lipogenic factors. Overexpression of ERK1/2 increased the expression of lipogenic genes, and orexin treatment induced the phosphorylated levels of ERK1/2Thr202/Tyr204, but not that of p38 Thr180/Tyr182 or JNK1/2 Thr183/Tyr185 in chicken liver and LMH cells. Taken together, this is the first report evidencing that orexin is expressed and secreted from chicken hepatocytes, and that orexin induced hepatic lipogenesis via activation of ERK1/2 signaling pathway.

Involvement of HO-1 and Autophagy in the Protective Effect of Magnolol in Hepatic Steatosis-Induced NLRP3 Inflammasome Activation In Vivo and In Vitro.

Magnolol (MG) is the main active compound of Magnolia officinalis and exerts a wide range of biological activities. In this study, we investigated the effects of MG using tyloxapol (Tylo)-induced (200 mg/kg, i.p.) hyperlipidemia in rats and palmitic acid (PA)-stimulated (0.3 mM) HepG2 cells. Our results showed that Tylo injection significantly increased plasma levels of triglyceride and cholesterol as well as superoxide anion in the livers, whereas MG pretreatment reversed these changes. MG reduced hepatic lipogenesis by attenuating sterol regulatory element-binding protein-1c (SREBP-1c) and fatty acid synthase (FAS) proteins and Srebp-1, Fas, Acc, and Cd36 mRNA expression as well as upregulated the lipolysis-associated genes Hsl, Mgl, and Atgl. Furthermore, MG reduced plasma interleukin-1β (IL-1β) and protein expression of NLR family pyrin domain-containing 3 (NLRP3), apoptosis-associated speck-like protein (ASC), and caspase 1 as well as upregulated nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) and induction of heme oxygenase-1 (HO-1) in hepatocytes of Tylo-treated rats. Enhanced autophagic flux by elevation of autophagy related protein 5-12 (ATG5-12), ATG7, Beclin1, and microtubule-associated protein light chain 3 B II (LC3BII)/LC3BI ratio, and reduction of sequestosome-1 (SQSTM1/p62) and phosphorylation of mTOR was observed by MG administration. However, autophagy inhibition with 3-methyladenine (3-MA) in HepG2 cells drastically abrogated the MG-mediated suppression of inflammation and lipid metabolism. In conclusion, MG inhibited hepatic steatosis-induced NLRP3 inflammasome activation through the restoration of autophagy to promote HO-1 signaling capable of ameliorating oxidative stress and inflammatory responses.

The Protective Effect of Myristica fragrans Houtt. Extracts Against Obesity and Inflammation by Regulating Free Fatty Acids Metabolism in Nonalcoholic Fatty Liver Disease.

Nonalcoholic fatty liver disease (NAFLD) is a disorder characterized by the excess accumulation of fat in the hepatocytes. It is commonly associated with severe obesity and inflammation. Free fatty acids (FFAs) are the key to regulate lipid metabolism and immune response in hepatocyte cells. This study examined the effects of AEN (alcohol extract of nutmeg, the seed of Myristica fragrans Houtt.) on the inhibition of lipid synthesis and inflammation in vitro and in vivo and on high-fat diet-induced obesity in NAFLD mice. Our results showed that AEN treatment could downregulate the expression of lipid synthesis-related genes fatty acid synthase (FASN) and sterol regulatory element-binding protein 1c (SREBP-1c) and lower the lipid content of cells. AEN also inhibited FFAs-mediated inflammation-related cytokines interleukin-6 (IL-6) and tumor necrosis factor α (TNFα) expression in cells. In a mouse model, AEN reduced the bodyweight of obese mice and improved NAFLD without affecting food intake. Further analysis revealed that AEN significantly reduced inflammation level, cholesterol and lipid accumulation, blood glucose, and other liver function indexes in mice fed with a high-fat diet. In conclusion, AEN inhibited the aggravation of obesity and inflammation by downregulating lipid-gene expression in the liver to ameliorate NAFLD.

Methylmercury displays pro-adipogenic properties in rainbow trout preadipocytes

Methylmercury (MeHg) is a ubiquitous contaminant largely found in aquatic environments, especially in species at high trophic level such as salmonids. The aim of this study was to evaluate the effects of MeHg on adipocyte differentiation and lipid metabolism in rainbow trout. Primary cultured preadipocytes were exposed to increasing concentrations of MeHg during six days with or without a hormonal cocktail. Main results showed a dose-dependent intracellular accumulation of neutral lipids with a preferential uptake of n-3 polyunsaturated fatty acids. Interestingly, this accumulation occurred after a fairly low uptake of MeHg by preadipocytes and was maintained after the cellular exposure to MeHg. In membrane phospholipids, arachidonic acid (20:4 n-6) was released in a dose-dependent manner. At the transcriptional level, the expression of several adipocyte-specific genes (perilipin 2 and apolipoprotein Eb) as well as lipid-related genes (fatty acid synthase and fatty acid binding protein 11a) was up-regulated in preadipocytes exposed to MeHg. These results highlight for the first time the disrupting effect of MeHg in trout adipocyte metabolism, providing new insights regarding the role of environmental pollutants in adipose tissue dysfunction and related pathologies.

Abstract 1899: Extracellular lipid starvation modulates the effects of BRAF inhibitors in melanoma

Abstract Melanoma is the most aggressive of all skin cancers, being the leading cause of death for cutaneous malignancy, with elevated metastatic spread and drug resistance. Because the deregulation of components of lipid metabolism may contribute to melanoma cell resistance, the aim of this study was to examine the dependence of melanoma cell growth on extracellular lipids, the associated lipid metabolism gene expression profiles and lipid impact on sensitivity to BRAF inhibitors. We applied preclinical pharmacology approaches including western blot analyses, growth inhibition assays quantitative Real-time PCR (qRT-PCR) in melanoma cell lines including variants with acquired resistance to BRAF inhibitors. Two matched pairs of sensitive and PLX4032-resistant cell lines (LM16 and LM36) were used. Both pairs carried V600E BRAF gene mutation and BRAF gene amplification, and LM36R displayed mutant NRAS. When compared to the parental sensitive cells, LM16R cells showed increased activation of MAPK-ERK and PI3K-Akt-mTOR pathways, while LM36R cells displayed increased activation of IGF1R and LKB1. Under standard culture conditions, when compared to the parental sensitive cells, both the resistant variants showed reduced expression of DHCR24 (24-Dehydrocholesterol Reductase), and LM16R showed reduced protein level of the lipogenic enzyme FASN (Fatty Acid Synthase) while LM36R cells displayed similar FASN protein levels. When grown in lipid-free medium, LM16 and LM16R cells increased the expression of both FASN and DHCR24 proteins. These changes were particularly evident for DHCR24 in LM16R cells after 48 h of growth in the absence of lipids. Lipid starvation markedly reduced cell growth of LM16-LM16R as well as LM36-LM36R pairs over time, albeit with a lower impact at higher cell density. Growth inhibition assays indicated that all the cells cultured in lipid-free medium were more sensitive to PLX4032 than those cultured in standard medium. An analysis of the expression of lipid metabolism genes by qRT-PCR showed that after 48 hours under lipid-free culture conditions the levels of FASN were slightly increased in LM16 and LM16R cells but not in the LM36 pair. Conversely, the mRNA levels of DHCR24 were enhanced upon lipid starvation only in LM16 and LM16R cell lines. Besides, under these conditions, HMGCR (3-Hydroxy-3-Methylglutaryl-CoA Reductase) mRNA levels increased in LM16, LM16R and LM36R cell lines. ACSS2 (Acetyl-coenzyme A synthetase 2) was increased particularly in the LM16 cells. These findings suggest that lipid starvation redirects lipid pathways toward cholesterol synthesis (acetate-cholesterol) by increasing ACSS2, HMGCR as well as DHCR24 levels. Thus, extracellular lipid availability may influence tumor cell response to treatment, a finding to be considered in the frame of a personalized therapy tailored to the specific characteristics of each patient. Citation Format: Giovanni L. Beretta, Elisabetta Vergani, Cristina Corno, Stella Tinelli, Monica Rodolfo, Laura Gatti, Paola Perego. Extracellular lipid starvation modulates the effects of BRAF inhibitors in melanoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1899.

Mixtures of Tea and Citrus maxima (pomelo) Alleviate Lipid Deposition in HepG2 Cells Through the AMPK/ACC Signaling Pathway.

Tea and citrus maxima are natural, medicinal homologous plants, typically used for making beverages, which have anticancer, antiobesity, and antioxidation properties. Green tea, yellow tea, and black tea were combined with citrus maxima to obtain green tea and Citrus maxima (GTCM), yellow tea and Citrus maxima (YTCM), and black tea and Citrus maxima (BTCM). The biochemical components of these mixtures were analyzed, and their possible effects and mechanisms on relieving liver lipid deposition were explored. The tea polyphenols, free amino acids, phenolamine ratio, and caffeine were comparable in YTCM and GTCM, being significantly higher than those in BTCM. In addition, the content of esterified catechins, nonesterified catechins, and total catechins in YTCM was significantly higher than those in GTCM and BTCM. All three mixtures of Citrus maxima tea significantly reduced lipid deposition in HepG2 cells, with GTCM and YTCM being slightly more effective than BTCM. Regarding the possible mechanism, Western blot analysis revealed that the three Citrus maxima tea mixtures could activate the AMPK/ACC signaling pathway, upregulate the expression of p-AMPK, p-ACC, and CPT-1 proteins, and downregulate the expression of SREBP1c and fatty acid synthase proteins to inhibit fat synthesis, thereby relieving lipid deposition in liver cells. In conclusion, as a novel and healthy beverage, Citrus maxima tea has the potential to alleviate liver lipid deposition, and further could be responsible for obesity treatment.