Fatty Acid Synthase mRNA Levels Research Articles

Salinity levels affect the lysine nutrient requirements and nutrient metabolism of juvenile genetically improved farmed tilapia (Oreochromis niloticus).

This 62-d research aimed to evaluate the effects of dietary lysine levels (DLL) and salinity on growth performance and nutrition metabolism of genetically improved farmed tilapia (GIFT) juveniles (Oreochromis niloticus). Six diets with lysine supplementation (1·34, 1·70, 2·03, 2·41, 2·72 and 3·04 % of DM) were formulated under different cultured salinities in a two-factorial design. The results indicated that supplemental lysine improved the specific growth rate (SGR) and weight gain (WG) and decreased the feed conversion ratio (FCR). Meanwhile, the fish had higher SGR and WG and lower FCR at 8 ‰ salinity. Except for moisture, the whole-body protein, lipid and ash content of GIFT were increased by 8 ‰ salinity, which showed that DLL (1·34 %) increased the whole-body fat content and DLL (2·41 %) increased whole-body protein content. Appropriate DLL up-regulated mRNA levels of protein metabolism-related genes such as target of rapamycin, 4EBP-1 and S6 kinase 1. However, 0 ‰ salinity reduced these protein metabolism-related genes mRNA levels, while proper DLL could improve glycolysis and gluconeogenesis mRNA levels but decrease lipogenesis-related genes mRNA levels in liver. 0 ‰ salinity improved GLUT2, glucokinase and G6 Pase mRNA levels; however, sterol regulatory element-binding protein 1 and fatty acid synthase mRNA levels were higher at 8 ‰ salinity. Moreover, 8 ‰ salinity also increased plasma total protein and cholesterol levels and decreased glucose levels. These results indicated that the recommended range of lysine requirement under different salinity was 2·03-2·20 % (0 ‰) and 2·20-2·41 % (8 ‰) and 8 ‰ salinity resulted in higher lysine requirements due to changes in the related nutrient metabolism, which might provide useful information for designing more effective feed formulations for GIFT cultured in different salinity environment.

Malonyl/Acetyltransferase (MAT) Knockout Decreases Triacylglycerol and Medium-Chain Fatty Acid Contents in Goat Mammary Epithelial Cells.

Malonyl/acetyltransferase (MAT) is a crucial functional domain of fatty acid synthase (FASN), which plays a vital role in the de novo synthesis of fatty acids in vivo. Milk fatty acids are secreted by mammary epithelial cells. Mammary epithelial cells are the units of mammary gland development and function, and it is a common model for the study of mammary gland tissue development and lactation. This study aimed to investigate the effects of MAT deletion on the synthesis of triacylglycerol and medium-chain fatty acids. The MAT domain was knocked out by CRISPR/Cas9 in the goat mammary epithelial cells (GMECs), and in MAT knockout GMECs, the mRNA level of FASN was decreased by approximately 91.19% and the protein level decreased by 51.83%. The results showed that MAT deletion downregulated the contents of triacylglycerol and medium-chain fatty acids (p < 0.05) and increased the content of acetyl-Coenzyme A (acetyl-CoA) (p < 0.001). Explicit deletion of MAT resulted in significant drop of FASN, which resulted in downregulation of LPL, GPAM, DGAT2, PLIN2, XDH, ATGL, LXRα, and PPARγ genes in GMECs (p < 0.05). Meanwhile, mRNA expression levels of ACC, FASN, DGAT2, SREBP1, and LXRα decreased following treatment with acetyl-CoA (p < 0.05). Our data reveals that FASN plays critical roles in the synthesis of medium-chain fatty acids and triacylglycerol in GMECs.

Effect of supplemental pantothenic acid on lipid metabolism and antioxidant function of Apis mellifera worker bees

This study was conducted to investigate the effects of pantothenic acid (PA) on the lipid metabolism, and antioxidant function of Apis mellifera workers. The results showed that ether extract (EE) content of bees’ body was significantly decreased when workers were supplemented with 147 mg/kg PA (P < 0.05). Further, supplemental PA reduced triglycerides (TG) and total cholesterol (TCHO) content in the hemolymph (P < 0.05), while 39.2 mg/kg PA supplementation improved hemolymph high-density lipoprotein cholesterol (HDL-c) concentration, while 261.6 mg/kg PA reduced low-density lipoprotein cholesterol (LDL-c) concentration (P < 0.05). C12:0, C14:1, and C18:0 content values were notably increased in PA-supplemented groups, while those of C14:0, C18:2n-6, C20:4n-6, polyunsaturated fatty acids (PUFA), and PUFA/saturated fatty acids were decreased (P < 0.05), while levels correlated with those of dietary PA. Malondialdehyde (MDA) content showed a downward trend in response to PA supplementation; however, total superoxide dismutase (T-SOD) was increased (P < 0.05). Furthermore, fatty acid synthase (FAS) mRNA levels were significantly downregulated in bees supplemented 9.8, 19.6, and 39.2 mg/kg (P < 0.05) and lipase activity significantly improved following supplementation with 39.2, 78.4, and 147 mg/kg PA (P < 0.05). Thus, dietary PA significantly influenced worker bee lipid metabolism, and improved antioxidant function.

Soyasaponin ameliorates obesity and reduces hepatic triacylglycerol accumulation by suppressing lipogenesis in high-fat diet-fed mice.

Soyasaponins are triterpenoid glycosides found in soybean. We investigated whether soyasaponin ameliorates lipid metabolism and its possible mechanisms. In C57BL/6J mice fed a high-fat diet (HFD), soyasaponin (SAP) was orally administered for 9weeks. Additionally, we evaluated the effect of soyasapogenols on 3T3-L1 adipocytes. In HFD-fed mice, the SAP significantly reduced body weight by 7% and relative adipose tissue weight by 35%. X-ray computed tomography demonstrated that the SAP reduced visceral and subcutaneous adipose tissue weights during week 3 of feeding. The SAP reduced sterol regulatory element-binding protein-1c (SREBP-1c) mRNA levels by 32% in the epididymal adipose tissue, significantly decreasing the triacylglycerol (TAG) content by 37% and SREBP-1c and fatty acid synthase mRNA levels by 52% and 61%, respectively, in the liver. In 3T3-L1 adipocytes, soyasapogenol B significantly decreased lipid droplets. The SAP containing soyasaponin A and B as conjugates demonstrate anti-obesity effects by suppressing adipocyte differentiation and lipogenesis, with a preventive effect on hepatic TAG accumulation by suppressing lipogenesis. PRACTICAL APPLICATION: Soyasaponin is one of the oleanane triterpenoids in soybeans. We have demonstrated that soyasaponin potently reduces body weight and white adipose tissue weight, and hepatic triacylglycerol accumulation in high-fat diet-fed mice. Thus, soyasaponin is a beneficial compound to prevent obesity and fatty liver.

Reducing FASN expression sensitizes acute myeloid leukemia cells to differentiation therapy

Fatty acid synthase (FASN) is the only human lipogenic enzyme available for de novo fatty acid synthesis and is often highly expressed in cancer cells. We found that FASN mRNA levels were significantly higher in acute myeloid leukemia (AML) patients than in healthy granulocytes or CD34+ hematopoietic progenitors. Accordingly, FASN levels decreased during all-trans retinoic acid (ATRA)-mediated granulocytic differentiation of acute promyelocytic leukemia (APL) cells, partially via autophagic degradation. Furthermore, our data suggest that inhibition of FASN expression levels using RNAi or (-)-epigallocatechin-3-gallate (EGCG) accelerated the differentiation of APL cell lines and significantly re-sensitized ATRA refractory non-APL AML cells. FASN reduction promoted translocation of transcription factor EB (TFEB) to the nucleus, paralleled by activation of CLEAR network genes and lysosomal biogenesis. Together, our data demonstrate that inhibition of FASN expression in combination with ATRA treatment facilitates granulocytic differentiation of APL cells and may extend differentiation therapy to non-APL AML cells.

Culture salinity alters dietary protein requirement, whole body composition and nutrients metabolism related genes expression in juvenile Genetically Improved Farmed Tilapia (GIFT) (Oreochromis niloticus)

An 8-week experimental trial was designed to explore the effects of dietary protein level (DPL) and salinity on the growth performance and physiological parameters of genetically improved farmed tilapia (GIFT) juveniles (Oreochromis niloticus). Six diets were formulated with different dietary protein levels (18.20%, 24.32%, 30.60%, 36.42%, 43.27% and 49.49%) and two salinity levels (0and 8‰) in a 6 × 2 factorial design. 720GIFT juveniles (initial weight 3.25 g) were distributed into 36 plastic tanks (180 L) and fed each of the diets four times daily to satiation. The results suggested that the growth performance was increased significantly with increasing DPLs up to a certain level (36.42%) in freshwater and brackish water. The whole-body lipid content of the fish decreased significantly with increasing salinity, while the protein content increased (P < 0.05). However, the protein content increased significantly and the lipid, moisture and ash contents were not significantly affected by the increase in DPL. With the increasing DPL at two salinities, the expression levels of hepatic insulin-like growth factor-1 increased significantly, reaching a maximum at 36.42% DPL. With the increasing DPL at the two salinity levels, the expression level of oligopeptide transporter 2 mRNA significantly decreased (P < 0.05). The target of rapamycin, 4E binding protein 1, ribosomal protein S6 and oligopeptide transporter 1β mRNA levels were not significantly affected by the DPL, but they increased significantly with an increase in salinity (P < 0.05). Regarding glucolipid metabolism, glycogen synthase, glucokinase and glucose transporter 2 mRNA levels decreased with an increase in DPL (P < 0.05). The fatty acid synthase and glucokinase mRNA levels decreased with an increase in salinity, while glycogen synthase and glucose transporter 2 mRNA levels increased (P < 0.05). The phosphoenolpyruvate carboxykinase mRNA level was not significantly affected by the DPL, but it increased with increasing salinity (P < 0.05). For the feed conversion ratio and weight gain rate, the broken-line model analysis showed that the optimal protein requirements at 8‰ salinity were higher than those at 0‰ salinity and the protein requirement were 37.70%, 37.86% (8‰) and 33.70%, 38.21% (0‰), respectively. GIFT juveniles exhibited improved glycogen synthesis at 8‰ salinity and fat deposition at 0‰ salinity.

Low-dose Bisphenol-A regulates inflammatory cytokines through GPR30 in mammary adipose cells.

The dramatic rise in obesity and metabolic syndrome can be related, at least in part, to environmental chemical factors such as Bisphenol-A (BPA). In this study, we aimed to understand the effects of low-dose Bisphenol-A on the human mature adipocytes and stromal vascular fraction (SVF) cells, obtained from subcutaneous mammary adipose tissue of overweight female patients, undergoing surgical mammary reduction. 24 and/or 48-h exposure to BPA 0.1 nM elicited significant increase of the inflammatory molecules interleukin-6 (IL-6), interleukin-8 (IL-8), monocyte chemo-attractant protein 1α (MCP1α) and induced G protein-coupled estrogen receptor 30 (GPR30) levels more than two-fold both in mature adipocytes and SVF cells. These effects were similar to that obtained in the presence of GPR30-specific agonist G1 (100 nM) and were reverted by G15 (1 µM), a GPR30-selective antagonist. As a result of BPA-GPR30 signaling activation, fatty acid synthase (FAS) and leptin mRNA levels were significantly higher upon BPA exposure (P < 0.05) in mature adipocytes, with an opposite effect on adiponectin (ADIPOQ). In addition, an increase in SVF cell proliferation and ERK1/2 phosphorylation, was observed, compared to untreated cells. G15 reverted all of these effects. Interestingly, the action of BPA on SVF cell growth was mimicked by IL-8 treatment and was reverted by incubation with anti-IL8 antibodies. All these data suggest that BPA at 0.1 nM, a ten times lower concentration than environmental exposure, increases the expression of pro-inflammatory cytokines via GPR30 both in mature mammary adipocytes and in SVF cells with a possible involvement of IL-8.

Differential effects of red yeast rice, Berberis aristata and Morus alba extracts on PCSK9 and LDL uptake

Background and aimsThe novel nutraceutical combination containing red yeast rice (monacolin K 3.3 mg), Berberis aristata cortex extract (Berberine 531.25 mg) and Morus alba leaves extract (1-deoxynojirimycin 4 mg) is effective in the management of elevated plasma low-density lipoprotein cholesterol (LDL-C) levels. The aim of the present study was to investigate the effects of the three components on proprotein convertase subtilisin/kexin type 9 (PCSK9), a key regulator of LDL receptor (LDLR) expression, in hepatocyte cell lines and to compare their effects on LDL cellular uptake. Methods and resultsHepG2 and Huh7 cells were incubated with B. aristata cortex extract (BCE), red yeast rice (RYR) and M. alba leaves extract (MLE) alone or in combination for 24 h. RYR (50 μg/mL) increased PCSK9 protein expression (Western blot analysis and ELISA), PCSK9 mRNA (qPCR) and its promoter activity (luciferase reporter assay). BCE (40 μg/mL) reduced instead PCSK9 expression, mRNA levels and promoter activity. MLE determined a concentration-dependent reduction of PCSK9 at the mRNA and protein levels, with a maximal reduction at 1 mg/mL, without significant changes of PCSK9 promoter activity. MLE also downregulated the expression of 3-hydroxy-3-methyl-3-glutaryl coenzyme A reductase and fatty acid synthase mRNA levels. The combination of RYR, BCE and MLE reduced the PCSK9 mRNA and protein levels, as well as the promoter activity. Finally, the single components and their combination induced LDL receptor and LDL uptake by the hepatocytes. ConclusionThe positive effect of MLE on PCSK9 supports the rationale of using the nutraceutical combination of RYR, BCE and MLE to control hyperlipidemic conditions.

Myricetin Ameliorates Ethanol-Induced Lipid Accumulation in Liver Cells by Reducing Fatty Acid Biosynthesis.

Alcoholic liver disease is a serious threat to human health. The development of drug candidates from complementary and alternative medicines is an attractive approach. Myricetin can be found in fruit, vegetables, and herbs. This study investigates the protective effect of myricetin on ethanol-induced injury in mouse liver cells. Oil-red O staining, assays of oxidative stress and measurements of inflammatory markers in mouse AML12 liver cells collectively demonstrate that myricetin elicits a curative effect on ethanol-induced injury. Next, the role of myricetin in the metabolic regulation of ethanol pathology in liver cells is assessed by gas chromatography coupled with mass spectrometry. Myricetin inhibits ethanol-stimulated fatty acid biosynthesis. Additionally, dodecanoic acid may be proposed as a potential biomarker related to ethanol pathology or myricetin therapy. It is also observed that myricetin enhances ethanol-induced inhibition of the mitochondrial electron transport chain. Moreover, fumaric acid is found to be a candidate biomarker related to ethanol toxicity or myricetin therapy. Quantitative reverse-transcription-PCR shows that ethanol-induced fatty acid synthase and sterol regulatory element-binding protein-1c mRNA levels are alleviated by myricetin. Finally, myricetin increases ethanol-induced inhibition of phosphorylation of AMP-activated protein kinase. These results elucidate the pharmacological mechanism of myricetin on ethanol-induced lipid accumulation.

Identification of metabolic pathways related to the bisphenol A-induced adipogenesis in differentiated murine adipocytes by using RNA-sequencing

We evaluated the effect of bisphenol A and its metabolites on the 3T3-L1 cells, in terms of glucose and lipid metabolism. We also aimed to obtain the information on the genome-wide expression changes in the 3T3-L1 cells treated with Bisphenol A by using RNA-seq, which involves whole-transcriptome sequencing. Differentially Expressed Genes (DEGs) collected from RNA-seq can be used to produce a complete picture of related metabolism pathways. The KEGG pathway was extracted based on the DEGs. Bisphenol A significantly increased the mRNA level of Sterol regulatory element binding transcription factor 1 (Srebf1) and CCAAT/enhancer binding protein alpha (Cebpa). Lipoprotein lipase (Lpl) was also significantly influenced by bisphenol A and its metabolites. Acetyl-Coenzyme A carboxylase beta (Acacb) and Fatty acid synthase (Fasn) mRNA levels were elevated by bisphenol A and its metabolites. The insulin signaling pathway, neurotrophin signaling pathway, and endometrial cancer-related pathway were focused by the functional enrichment analyses, and the pathways were well coincided with recent previous reports. DEGs collected from RNA-seq were confirmed as a reliable evidence in the exposure to the chemicals such as bisphenol A. Collecting pieces of the puzzles obtained from the RNA-seq will help us to produce a complete picture of the metabolic pathway for such chemicals.

Pex11a deficiency causes dyslipidaemia and obesity in mice.

Peroxisomes play a central role in lipid metabolism. We previously demonstrated that Pex11a deficiency impairs peroxisome abundance and fatty acid β‐oxidation and results in hepatic triglyceride accumulation. The role of Pex11a in dyslipidaemia and obesity is investigated here with Pex11a knockout mice (Pex11a−/−). Metabolic phenotypes including tissue weight, glucose tolerance, insulin sensitivity, cholesterol levels, fatty acid profile, oxygen consumption, physical activity were assessed in wild‐type (WT) and Pex11a−/− fed with a high‐fat diet. Molecular changes and peroxisome abundance in adipose tissue were evaluated through qRT‐PCR, Western blotting, and Immunofluorescence. Pex11a−/− showed increased fat mass, decreased skeletal muscle, higher cholesterol levels, and more severely impaired glucose and insulin tolerance. Pex11a−/− consumed less oxygen, indicating a decrease in fatty acid oxidation, which is consistent with the accumulation of very long‐ and long‐chain fatty acids. Adipose palmitic acid (C16:0) levels were elevated in Pex11a−/−, which may be because of dramatically increased fatty acid synthase mRNA and protein levels. Furthermore, Pex11a deficiency increased ventricle size and macrophage infiltration, which are related to the reduced physical activity. These data demonstrate that Pex11a deficiency impairs physical activity and energy expenditure, decreases fatty acid β‐oxidation, increases de novo lipogenesis and results in dyslipidaemia and obesity.

Adaptation of Nile tilapia (Oreochromis niloticus) to different levels of dietary carbohydrates: New insights from a long term nutritional study

The objective of this study was to examine the long-term effects of different levels of dietary carbohydrates on glucose metabolism in the omnivorous Nile Tilapia (Oreochromis niloticus). Fish were fed from the first feeding stage until the adult stage, over a period of 40 weeks, using three different levels of carbohydrates (dextrin): 0% (CHO-L), 30% (CHO-M) and 50% (CHO-H). Growth performance, blood metabolite parameters, the proximate composition of the whole body, muscle and liver tissue, and the mRNA levels for genes involved in glycolysis, gluconeogenesis, lipogenesis and glucose transport in liver and muscle tissue were analyzed at week 26 and week 40. Fish fed the CHO-M diet exhibited the best growth performance, while the fish fed with the CHO-H diet showed the lowest growth performance. These data suggest that the incorporation of a certain level dietary carbohydrates can benefit Nile Tilapia. However, if the proportion of carbohydrates is very high, and concomitantly, the level of dietary proteins too low, this will not promote growth. Fish fed with carbohydrates exhibited a significant increase in lipid deposition in the whole body, muscle and liver tissues in. This seems to be due to increased lipogenic capacity, as reflected by higher hepatic fatty acid synthase mRNA levels and activity, as well as higher levels of plasma triglyceride. Nile Tilapia appear to be able to effectively adapt to the intake of carbohydrates, as reflected by: (i) the weak postprandial hyperglycemia (5.3–6.1 mM), (ii) the higher level of hepatic glycogen, (iii) the higher glycolytic pyruvate kinase enzyme in muscle (mRNA level and activity) and (iv) the inhibition of the gluconeogenic pathway for glucose-6-phosphatase and phosphoenolpyruvate carboxykinase enzymes (at molecular and enzymatic levels). Overall, these data show that the well-known differences in the capacity of carnivorous and omnivorous fish species to use dietary carbohydrates could be linked to differences in the regulation of the gluconeogenic and lipogenic pathways.

Breeding selection of rainbow trout for high or low muscle adiposity differentially affects lipogenic capacity and lipid mobilization strategies to cope with food deprivation

The present study evaluates the transcriptional regulation of lipid metabolism, adiponectin system and oxidative stress genes in two rainbow trout lines selected over seven generations for low (lean line, LL) or high (fat line, FL) muscle adiposity, and subjected to different fasting regimes. Under feeding conditions, FL fish had higher muscle lipid content, plasma triglycerides (TG) and non-esterified fatty acids (NEFAs) than the LL fish. Besides, FL fish had higher fatty acid synthase (fasn) mRNA levels in adipose tissue (AT) and white muscle (WM), and higher lipoprotein lipase (lpl) mRNA levels in WM, suggesting an increased lipogenic capacity and fatty acid uptake in the fatty genotype. In response to fasting, the two fish lines displayed a different trend in muscle lipid mobilization, plasma TG levels changes and lipid metabolism-related genes expression patterns. There were different fasn expression profiles between the fish lines in both, AT and WM, in agreement with the changes observed in plasma TG levels. The FL fish had higher adiponectin receptor 2 mRNA levels in WM and adiponectin mRNA levels in AT than LL fish after 1 and 4 weeks of fasting respectively, indicating a difference between genotypes in the adiponectin system. Furthermore, oxidative stress genes mRNA levels in both fish lines showed a different pattern between AT and WM upon fasting, probably indicating a higher protective effect in WM. Overall, the present study reveals a distinct metabolic regulation for each genotype, highlighting their different strategies in response to food deprivation.

Involvement of glucocorticoid prereceptor metabolism and signaling in rat visceral adipose tissue lipid metabolism after chronic stress combined with high-fructose diet.

Both fructose overconsumption and increased glucocorticoids secondary to chronic stress may contribute to overall dyslipidemia. In this study we specifically assessed the effects and interactions of dietary fructose and chronic stress on lipid metabolism in the visceral adipose tissue (VAT) of male Wistar rats. We analyzed the effects of 9-week 20% high fructose diet and 4-week chronic unpredictable stress, separately and in combination, on VAT histology, glucocorticoid prereceptor metabolism, glucocorticoid receptor subcellular redistribution and expression of major metabolic genes. Blood triglycerides and fatty acid composition were also measured to assess hepatic Δ9 desaturase activity. The results showed that fructose diet increased blood triglycerides and Δ9 desaturase activity. On the other hand, stress led to corticosterone elevation, glucocorticoid receptor activation and decrease in adipocyte size, while phosphoenolpyruvate carboxykinase, adipose tissue triglyceride lipase, FAT/CD36 and sterol regulatory element binding protein-1c (SREBP-1c) were increased, pointing to VAT lipolysis and glyceroneogenesis. The combination of stress and fructose diet was associated with marked stimulation of fatty acid synthase and acetyl-CoA carboxylase mRNA level and with increased 11β-hydroxysteroid dehydrogenase type 1 and hexose-6-phosphate dehydrogenase protein levels, suggesting a coordinated increase in hexose monophosphate shunt and de novo lipogenesis. It however did not influence the level of peroxisome proliferator-activated receptor-gamma, SREBP-1c and carbohydrate responsive element-binding protein. In conclusion, our results showed that only combination of dietary fructose and stress increase glucocorticoid prereceptor metabolism and stimulates lipogenic enzyme expression suggesting that interaction between stress and fructose may be instrumental in promoting VAT expansion and dysfunction.

MiR-205-5p negatively regulates hepatic acetyl-CoA carboxylase β mRNA in lipid metabolism of Oreochromis niloticus

MicroRNAs (miRNAs) are non-coding RNAs that function as post-transcriptional gene regulators and that play vital roles controlling lipid metabolism. miR-205 is an important miRNA related to adipogenesis and lipid metabolism. However, little is known about the potential role of miR-205-5p in genetically improved farmed tilapia (GIFT, Oreochromis niloticus). In this study, we used miRanda software to search for potential miR-205-5p target genes and found a lipid-metabolism-related gene called acetyl-CoA carboxylase β (ACACβ). Quantitative real-time polymerase chain reaction data indicated that there may be a negative regulation relationship between miR-205-5p and ACACβ gene expression under HFD rearing. Using luciferase reporter assays, we verified the binding site of miR-205-5p in the 3′-untranslated region of the ACACβ mRNA. Furthermore, an in vivo functional analysis of miR-205-5p was performed by injecting GIFT juveniles with a miR-205-5p antagomir. Reduced levels of miR-205-5p in GIFT liver increased ACACβ mRNA expression 12 h post-injection. miR-205-5p suppression also increased fatty acid synthase and peroxisome proliferator-activated receptor-α mRNA levels 48 h and 120 h post-injection, respectively. Taken together, our results indicate that miR-205-5p negative regulates hepatic ACACβ mRNA expression, and may serve as an important regulator in controlling hepatic lipid metabolism in GIFT.

Interleukin-15 derived from Guanylin–GC-C-expressing macrophages inhibits fatty acid synthase in adipocytes

Recently we found that guanylin (Gn) and its receptor, guanylyl cyclase C (GC-C), are uniquely expressed in the mesenteric macrophages of some diet-resistant rats and that double-transgenic (dTg) rats overexpressing Gn and GC-C in macrophages demonstrate reduced fatty acid synthase and fat accumulation in fat tissue even when fed a high-fat diet (HFD). Lipid accumulation and fatty acid synthase mRNA levels in cocultured dTg rat adipocytes and macrophages were reduced compared with those in adipocytes cultured with WT rat macrophages. Here, we investigated whether Interleukin-15 (IL-15) derived from Gn–GC-C-expressing macrophages regulates lipid accumulation in adipocytes. IL-15 inhibited fatty acid synthase and lipid accumulation via STAT5 in cultured adipocytes. IL-15 mRNA and protein levels in the mesenteric fat of HFD-fed dTg rats were significantly higher than those of HFD-fed WT rats. Phosphorylated STAT5 levels in the mesenteric fat of HFD-fed dTg rats were increased compared with those of HFD-fed WT rats. In addition, the mRNA level of fatty acid synthase in the mesenteric fat was lower in HFD-fed dTg rats than in HFD-fed WT rats. These results support the hypothesis that IL-15 secreted from Gn–GC-C-expressing macrophages contributes to the inhibition of fatty acid synthase and lipid accumulation in adipocytes, leading to obesity resistance.

Environmental exposure to a major urban wastewater effluent: Effects on the energy metabolism of northern pike

Municipal wastewater effluents (MWWEs) consist of dynamic and complex mixtures of chemical and biological compounds that can alter the health of exposed aquatic organisms. Disturbance of energy metabolism has been reported in fish exposed to MWWEs. However, there is a scarcity of knowledge on the physiological events leading to perturbation of energy balance and thyroid regulation, and associated lipid metabolism. The objective of the present study was to use a set of biomarkers, from gene transcription to body condition, to investigate the effects of a chronic environmental exposure to a major primary MWWE on fatty acid metabolism and thyroid hormone levels in northern pike (Esox lucius) collected from the St. Lawrence River near Montreal (QC, Canada). The exposure of pike to MWWE was examined through determination of a suite of persistent and bioaccumulative halogenated flame retardants in liver as this effluent is a known regional source for these chemicals. Greater hepatic concentrations of polybrominated diphenyl ethers (PBDEs, range: 29.6–465ng/g w.w. and 88.8–823ng/g w.w. in females and males, respectively) and other halogenated flame retardants (e.g., dechlorane-related compounds) were determined in fish collected downstream of the MWWE’s point of discharge relative to the upstream site. This exposure in male pike was associated with decreased acyl-coA oxidase (acox1) and fatty acid synthase (fasn) mRNA levels as well as a decreased acyl-coA oxidase (ACOX) activity in liver. In female pike, MWWE exposure was associated with lower circulating free and total triiodothyronine (T3) levels and a tendency for greater total lipid percentages in liver. Present findings provide evidence that chronic exposure of a top predator fish to MWWE can be related to gender-specific effects on fatty acid metabolism and thyroid hormone homeostasis, and highlight the need for further investigation.

The additive effects of the TM6SF2 E167K and PNPLA3 I148M polymorphisms on lipid metabolism.

There is a genetic susceptibility for nonalcoholic fatty liver disease (NAFLD). To examine the role of genetic factors in the disease, a Bayesian analysis was performed to model gene relationships in NAFLD pathogenesis. The Bayesian analysis indicated a potential gene interaction between the TM6SF2 and PNPLA3 genes. Next, to explore the underlying mechanism at the cellular level, we evaluated the additive effects between the TM6SF2 E167K and PNPLA3 I148M polymorphisms on lipid metabolism. Hepa 1-6 cells were transfected with a control vector or with overexpression vectors for TM6SF2/PNPLA3-wild type, TM6SF2-mutant type, PNPLA3-mutant type, or TM6SF2/PNPLA3-mutant type. Commercial kits were used to measure triglyceride and total cholesterol levels in each of the five groups. The mRNA and protein expression levels of sterol regulatory element-binding transcription factor 1c and fatty acid synthase were analyzed using real-time PCR and western blotting. The triglyceride and total cholesterol contents were significantly different among the groups. The triglyceride and total cholesterol contents and the sterol regulatory element-binding transcription factor 1c and fatty acid synthase mRNA and protein expression levels were significantly higher in the TM6SF2/PNPLA3-mutant type group than in the TM6SF2-mutant type group or the PNPLA3-mutant type group. The TM6SF2 E167K and PNPLA3 I148M polymorphisms may have additive effects on lipid metabolism by increasing the expression of sterol regulatory element-binding transcription factor 1c and fatty acid synthase.

Tributyltin and triphenyltin exposure promotes in vitro adipogenic differentiation but alters the adipocyte phenotype in rainbow trout

Numerous environmental pollutants have been identified as potential obesogenic compounds affecting endocrine signaling and lipid homeostasis. Among them, well-known organotins such as tributyltin (TBT) and triphenyltin (TPT), can be found in significant concentrations in aquatic environments. The aim of the present study was to investigate in vitro the effects of TBT and TPT on the development and lipid metabolism of rainbow trout (Onchorynchus mykiss) primary cultured adipocytes. Results showed that TBT and TPT induced lipid accumulation and slightly enhanced peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT enhancer binding protein alpha (C/EBPα) protein expression when compared to a control, both in the presence or absence of lipid mixture. However, the effects were higher when combined with lipid, and in the absence of it, the organotins did not cause complete mature adipocyte morphology. Regarding gene expression analyses, exposure to TBT and TPT caused an increase in fatty acid synthase (fasn) mRNA levels confirming the pro-adipogenic properties of these compounds. In addition, when added together with lipid, TBT and TPT significantly increased cebpa, tumor necrosis factor alpha (tnfa) and ATP-binding cassette transporter 1 (abca1) mRNA levels suggesting a synergistic effect. Overall, our data highlighted that TBT and TPT activate adipocyte differentiation in rainbow trout supporting an obesogenic role for these compounds, although by themselves they are not able to induce complete adipocyte development and maturation suggesting that these adipocytes might not be properly functional.

Dietary intervention, but not losartan, completely reverses non-alcoholic steatohepatitis in obese and insulin resistant mice

BackgroundDietary intervention is the cornerstone of non-alcoholic steatohepatitis (NASH) treatment. However, histological evidence of its efficacy is limited and its impact on hepatic pathways involved in NASH is underreported. The efficacy of the angiotensin receptor type 1 blocker losartan is controversial because of varying results in a few animal and human studies. We evaluated the effect of dietary intervention versus losartan on NASH and associated systemic metabolic features in a representative mouse model.MethodsMale C57BL/6 J mice with high fat-high sucrose diet (HF-HSD) induced NASH, obesity, insulin resistance and hypercholesterolemia were subjected to dietary intervention (switch from HF-HSD to normal chow diet (NCD)) (n = 9), continuation HF-HSD together with losartan (30 mg/kg/day) (n = 9) or continuation HF-HSD only (n = 9) for 8 weeks. 9 mice received NCD during the entire experiment (20 weeks). We assessed the systemic metabolic effects and performed a detailed hepatic histological and molecular profiling. A P-value of < 0.05, using the group with continuation of HF-HSD only as control, was considered as statistically significant.ResultsDietary intervention normalized obesity, insulin resistance, and hypercholesterolemia (for all P < 0.001), and remarkably, completely reversed all histological features of pre-existent NASH (for all P < 0.001), including fibrosis measured by quantification of collagen proportional area (P < 0.01). At the hepatic molecular level, dietary intervention targeted fibrogenesis with a normalization of collagen type I alpha 1, transforming growth factor β1, tissue inhibitor of metalloproteinase 1 mRNA levels (for all P < 0.01), lipid metabolism with a normalization of fatty acid translocase/CD36, fatty acid transport protein 5, fatty acid synthase mRNA levels (P < 0.05) and markers related to mitochondrial function with a normalization of hepatic ATP content (P < 0.05) together with sirtuin1 and uncoupling protein 2 mRNA levels (for both P < 0.001). Dietary intervention abolished p62 accumulation (P < 0.01), suggesting a restoration of autophagic flux. Losartan did not significantly affect obesity, insulin resistance, hypercholesterolemia or any histological NASH feature.ConclusionsDietary intervention, and not losartan, completely restores the metabolic phenotype in a representative mouse model with pre-existent NASH, obesity, insulin resistance and hypercholesterolemia.