Lipid Synthesis-related Genes Research Articles

Identification of key genes for triacylglycerol biosynthesis and storage in herbaceous peony (Paeonia lactifolra Pall.) seeds based on full-length transcriptome

BackgroundThe herbaceous peony (Paeonia lactiflora Pall.) is extensively cultivated in China due to its root being used as a traditional Chinese medicine known as ‘Radix Paeoniae Alba’. In recent years, it has been discovered that its seeds incorporate abundant unsaturated fatty acids, thereby presenting a potential new oilseed plant. Surprisingly, little is known about the full-length transcriptome sequencing of Paeonia lactiflora, limiting research into its gene function and molecular mechanisms.ResultsA total of 484,931 Reads of Inserts (ROI) sequences and 1,455,771 full-Length non-chimeric reads (FLNC) sequences were obtained for CDS prediction, TF analysis, SSR analysis and lncRNA identification. In addition, gene function annotation and gene structure analysis were performed. A total of 4905 transcripts were related to lipid metabolism biosynthesis pathway, belonging to 28 enzymes. We use these data to identify 10 oleosin (OLE) and 5 diacylglycerol acyltransferase (DGAT) gene members after de-redundancy. The analysis of physicochemical properties and secondary structure showed them similarity in gene family respectively. The phylogenetic analysis showed that the distribution of OLE and DGAT family members was roughly the same as that of Arabidopsis. Quantitative real-time polymerase chain reaction (qRT–PCR) analyses revealed expression changes in different seed development stages, and showed a trend of increasing and then decreasing.ConclusionIn summary, these results provide new insights into the molecular mechanism of triacylglycerol (TAG) biosynthesis and storage during the seedling stage in Paeonia lactiflora. It provides theoretical references for selecting and breeding oil varieties and understanding the functions of oil storage as well as lipid synthesis related genes in Paeonia lactiflora.

Ism1 deficiency in mice exacerbates bleomycin-induced pulmonary fibrosis with enhanced cellular senescence and delayed fibrosis resolution

Idiopathic pulmonary fibrosis (IPF) is a progressive fibrosing interstitial lung disease with high morbidity and mortality but unclear etiology and incomplete pathophysiological understandings, making the discovery of effective therapeutics arduous. Currently, two drugs, Nintedanib and Pirfenidone, are available for IPF treatment which can slow down the fibrotic scarring in the lung but are unable to provide disease resolution. Hence, further revelation of the molecular mechanisms of IPF is critical for the development of novel therapeutics. Isthmin-1 (ISM1) is a secreted anti-inflammatory protein highly expressed in the mouse and human lung. Ism1−/− mice presented spontaneous and progressive lung emphysema, as well as heightened acute lung injury (ALI) upon lipopolysaccharide (LPS) treatment with an accompanied increase of post-LPS-ALI pulmonary fibrosis. ISM1 is important for lung homeostasis with airway-delivered recombinant ISM1 (rISM1) suppressing cigarette smoke-induced emphysema, LPS-ALI, and house-dust mites (HDM)-induced asthma-like symptoms in mice. However, the role of ISM1 in pulmonary fibrosis is yet to be clearly understood. In this work, we show that Ism1−/− mice presented heightened bleomycin-induced pulmonary fibrosis (BIPF), with enhanced immune cell infiltration, myofibroblast accumulation, and collagen deposition. ISM1 deficiency also led to increased cellular senescence in mouse lungs, isolated primary alveolar type II epithelial cells, and primary lung fibroblasts upon bleomycin treatment. Ism1−/− mice also showed delayed resolution of pulmonary fibrosis with reduced lipofibroblasts and downregulation of lipid synthesis-related genes. These results are in congruence with the RNA-seq data which demonstrated gene expression alterations in Ism1−/− lung that are linked to predisposition to pulmonary fibrosis and dysregulation of lipid metabolism pathways. Gene expression analyses showed that Ism1 is similarly dysregulated in the lungs of BIPF and human IPF patients. These findings reveal an anti-fibrotic role of ISM1 in mouse lungs and provide the foundation to further investigate possible therapeutic applications of ISM1 for pulmonary fibrosis in the future.

#1957 Microbial and metabolomic remodeling by Sichuan dark tea-based formula improves obesity-induced renal lipid metabolism disorder in mice

Abstract Background and Aims The theory of medicine-food homology is long-standing and has been widely applied in traditional Chinese medicine. The Sichuan dark tea-based medicated dietary formula (alternatively referred to as Qing, or clarity in Chinese) is previously used for its lipid-lowering properties in apoE mice. This study further investigated the renoprotective effects of Qing on diet-induced obesity (DIO) mice and explored the specific mechanisms involved. Method Male C57BL/6 mice were randomly assigned to three groups, a control group, a DIO group, and a Qing treatment group, or the Qing group, with 8 mice in each group. The mice in the control group were given normal maintenance feed and purified water, and the other two groups were fed a high-fat diet for 12 weeks to establish the DIO model. After that, high-fat diet continued in the DIO group, while the Qing group was given Qing at the same time for 12 weeks, during which period the weight of the mice was monitored and recorded every week. The mice were sacrificed after 12 weeks. Serum samples were collected and the levels of triglyceride (TG), total cholesterol (TC), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and albumin were measured to evaluate liver function. In addition, renal lipids were extracted to determine the levels of TG and TC in the kidney and periodic acid-Schiff (PAS) and oil red O stainings were performed to evaluate kidney pathological injury. Western blot was performed to determine the phosphorylated AMPK (pAMPK)/AMPK ratio in the kidney tissue. RT-qPCR and Western blot were used to determine the expression of proteins related to fatty acid oxidation and key proteins related to lipid synthesis. 16SrRNA and metabolomics were applied to analyze the gut microbiota in the intestinal contents and its metabolites. Results Compared with those of the control group, the levels of liver mass (P=0.0003), serum ALT(P < 0.0001), AST (P=0.0001), TC (P=0.0191) and TG (P=0.0101) in the model group were significantly increased. Qing treatment can effectively reduce liver mass (P=0.0316), and improve AST (P=0.0012), ALT (P=0.0027), TC (P=0.0200), TG (P=0.0499) levels of mice compared to those of the DIO group. Qing treatment inhibited the expression of lipid synthesis related genes (SREBP-1, FASN and SCD1) and promoted the expression of fatty acid oxidation related genes (CPT1A, PGC1α and PPARγ) in kidneys (Fig. 1). Obesity-triggered dysfunction of structure and composition of gut microbiota were improved by Qing (Fig. 2), where the content of short-chain fatty acids, especially isovaleric acid and propionic acid, were also restored (Fig. 3). Conclusion Our data suggested that Sichuan dark tea-based medicated dietary formula may improve renal lipid metabolism by regulating gut microbiota and the levels of intestinal short-chain fatty acids, thereby protecting obesity-related kidney injury. Isovaleric acid and propionic acid may be the metabolites key to its regulation of gut microbiota.

Identifying differentially expressed genes in goat mammary epithelial cells induced by overexpression of SOCS3 gene using RNA sequencing.

The suppressor of cytokine signaling 3 (SOCS3) is a key signaling molecule that regulates milk synthesis in dairy livestock. However, the molecular mechanism by which SOCS3 regulates lipid synthesis in goat milk remains unclear. This study aimed to screen for key downstream genes associated with lipid synthesis regulated by SOCS3 in goat mammary epithelial cells (GMECs) using RNA sequencing (RNA-seq). Goat SOCS3 overexpression vector (PC-SOCS3) and negative control (PCDNA3.1) were transfected into GMECs. Total RNA from cells after SOCS3 overexpression was used for RNA-seq, followed by differentially expressed gene (DEG) analysis, functional enrichment analysis, and network prediction. SOCS3 overexpression significantly inhibited the synthesis of triacylglycerol, total cholesterol, non-esterified fatty acids, and accumulated lipid droplets. In total, 430 DEGs were identified, including 226 downregulated and 204 upregulated genes, following SOCS3 overexpression. Functional annotation revealed that the DEGs were mainly associated with lipid metabolism, cell proliferation, and apoptosis. We found that the lipid synthesis-related genes, STAT2 and FOXO6, were downregulated. In addition, the proliferation-related genes BCL2, MMP11, and MMP13 were upregulated, and the apoptosis-related gene CD40 was downregulated. In conclusion, six DEGs were identified as key regulators of milk lipid synthesis following SOCS3 overexpression in GMECs. Our results provide new candidate genes and insights into the molecular mechanisms involved in milk lipid synthesis regulated by SOCS3 in goats.

Pleurotus eryngii root waste and soybean meal co-fermented protein improved the growth, immunity, liver and intestinal health of largemouth bass (Micropterus salmoides)

The present study aimed to evaluate the effect of king oyster mushroom (Pleurotus eryngii) root waste and soybean meal co-fermented protein (CFP) on growth performance, feed utilization, immune status, hepatic and intestinal health of largemouth bass (Micropterus salmoides). Largemouth bass (12.33 ± 0.18 g) were divided into five groups, fed with diets containing 0 %, 5 %, 10 %, 15 % and 20 % CFP respectively for 7 weeks. The growth performance and dietary utilization were slightly improved by the supplementation of CFP. In addition, improved immunoglobulin M (IgM) content and lysozyme activity in treatments confirm the enhancement of immunity in fish by the addition of CFP, especially in fish fed 20 % CFP (P < 0.05). Furthermore, CFP significantly improved liver GSH (glutathione) content in groups D10 and D15 (P < 0.05), and slightly improved total antioxidant capacity (T-AOC), superoxide dismutase (SOD) activity while slightly reduced malondialdehyde (MDA) content. Simultaneously, the upregulation of lipolysis-related genes (PPARα, CPT1 and ACO) expression and downregulation of lipid synthesis-related genes (ACC and DGAT1) expression was recorded in the group D20 compared with the control (P < 0.05), which were consistent with the decreased liver lipid contents, suggests that lipid metabolism was improved by CFP. In terms of intestinal structural integrity, ameliorated intestinal morphology in treatments were consistent with the upregulated Occludin, Claudin-1 and ZO-1 genes expression. The intestinal pro-inflammatory cytokines (TNF-α and IL-8) expression were suppressed while the anti-inflammatory cytokines (IL-10 and TGF-β) were activated in treatments. The expression of antimicrobial peptides (Hepcidin-1, Piscidin-2 and Piscidin-3) and intestinal immune effectors (IgM and LYZ) were slightly up-regulated in treatments. Additionally, the relative abundance of intestinal beneficial bacteria (Firmicutes) increased while the relative abundance of potential pathogenic bacteria (Fusobacterium and Proteobacteria) decreased, which indicated that the intestinal microbial community was well-reorganized by CFP. In conclusion, dietary CFP improves growth, immunity, hepatic and intestinal health of largemouth bass, these data provided a theoretical basis for the application of this novel functional protein ingredient in fish.

Effects of nuciferine on growth performance, body composition, lipid metabolism and antioxidant capacity of juvenile large yellow croaker (Larmichthys crocea) fed diets with high palm oil level

This study was conducted to explore the impacts of nuciferine (NF) on growth performance, body composition, lipid metabolism and antioxidant capacity of juvenile large yellow croaker (Larmichthys crocea) fed diets with high palm oil level. Five diets were formulated with fish oil (FO), palm oil (PO) and PO supplemented with 0.01%NF, 0.05%NF and 0.25%NF. After feeding for 10 weeks, the results showed that the growth performance (final body weight and specific growth rate) exhibited a significantly quadratic trend with the increase of NF addition, and the maximums were observed in PO supplemented with 0.05%NF group. Moreover, dietary PO diet with NF addition relieved the abnormal lipid accumulation (hepatosomatic index, crude lipid and triglyceride contents, total cholesterol contents, low-density lipoprotein-cholesterol contents in liver and serum) induced by PO diet. Simultaneously, NF addition inhibited PO diet-induced increase in the mRNA expressions of lipid synthesis-related genes and PO diet-induced activation of SREBP1. In addition, dietary PO diet with NF addition impeded PO diet-induced increase of hepatic MDA contents and recovered PO diet-induced inhibition of SOD and CAT activities. Similarly, NF addition alleviated PO diet-induced decrease in the mRNA expressions of antioxidant-related genes and PO diet-induced inhibition of SOD protein expression. Thus, addition of NF in PO diet might effectively ameliorate PO diet-induced growth retardation, lipid metabolism disorder and oxidative stress in croaker, and the optimal addition dose of NF in PO diet was approximately 0.05%.

Lipid homeostasis is essential for oogenesis and embryogenesis in the silkworm, Bombyx mori

Reproduction, a fundamental feature of all known life, closely correlates with energy homeostasis. The control of synthesizing and mobilizing lipids are dynamic and well-organized processes to distribute lipid resources across tissues or generations. However, how lipid homeostasis is precisely coordinated during insect reproductive development is poorly understood. Here we describe the relations between energy metabolism and reproduction in the silkworm, Bombyx mori, a lepidopteran model insect, by using CRISPR/Cas9-mediated mutation analysis and comprehensively functional investigation on two major lipid lipases of Brummer (BmBmm) and hormone-sensitive lipase (BmHsl), and the sterol regulatory element binding protein (BmSrebp). BmBmm is a crucial regulator of lipolysis to maintain female fecundity by regulating the triglyceride (TG) storage among the midgut, the fat body, and the ovary. Lipidomics analysis reveals that defective lipolysis of females influences the composition of TG and other membrane lipids in the BmBmm mutant embryos. In contrast, BmHsl mediates embryonic development by controlling sterol metabolism rather than TG metabolism. Transcriptome analysis unveils that BmBmm deficiency significantly improves the expression of lipid synthesis-related genes including BmSrebp in the fat body. Subsequently, we identify BmSrebp as a key regulator of lipid accumulation in oocytes, which promotes oogenesis and cooperates with BmBmm to support the metabolic requirements of oocyte production. In summary, lipid homeostasis plays a vital role in supporting female reproductive success in silkworms.

PEDV inhibits HNRNPA3 expression by miR-218-5p to enhance cellular lipid accumulation and promote viral replication.

Porcine epidemic diarrhea virus (PEDV) requires complete dependence on the metabolic system of the host cell to complete its life cycle. There is a strong link between efficient viral replication and cellular lipid synthesis. However, the mechanism by which PEDV interacts with host cells to hijack cellular lipid metabolism to promote its replication remains unclear. In this study, PEDV infection significantly enhanced the expression of lipid synthesis-related genes and increased cellular lipid accumulation. Furthermore, using liquid chromatography-tandem mass spectrometry, we identified heterogeneous nuclear ribonucleoprotein A3 (HNRNPA3) as the interacting molecule of PEDV NSP9. We demonstrated that the expression of HNRNPA3 was downregulated by PEDV-induced miR-218-5p through targeting its 3' untranslated region. Interestingly, knocking down HNRNPA3 facilitated the PEDV replication by promoting cellular lipid synthesis. We next found that the knockdown of HNRNPA3 potentiated the transcriptional activity of sterol regulatory element-binding transcription factor 1 (SREBF1) through zinc finger protein 135 (ZNF135) as well as PI3K/AKT and JNK signaling pathways. In summary, we propose a model in which PEDV downregulates HNRNPA3 expression to promote the expression and activation of SREBF1 and increase cellular lipid accumulation, providing a novel mechanism by which PEDV interacts with the host to utilize cellular lipid metabolism to promote its replication.IMPORTANCEAs the major components and structural basis of the viral replication complexes of positive-stranded RNA viruses, lipids play an essential role in viral replication. However, how PEDV manipulates host cell lipid metabolism to promote viral replication by interacting with cell proteins remains poorly understood. Here, we found that SREBF1 promotes cellular lipid synthesis, which is essential for PEDV replication. Moreover, HNRNPA3 negatively regulates SREBF1 activation and specifically reduces lipid accumulation, ultimately inhibiting PEDV dsRNA synthesis. Our study provides new insight into the mechanisms by which PEDV hijacks cell lipid metabolism to benefit viral replication, which can offer a potential target for therapeutics against PEDV infection.

Response of Neuropeptides to Hunger Signals in Teleost

Introduction: The perception of hunger is a complex physiological process that requires precise coordination between the central and peripheral tissues. Methods: In this study, tilapia fasted for 24 h was chosen to establish a hunger model to study the mechanism of homeostasis recovery under the joint regulation of the central nervous system (CNS) and peripheral tissues. Results: The gastric and intestinal contents of tilapia were predominantly depleted after a fasting period of 9 h and 24 h, respectively. The serum glucose level significantly decreased at the 9-h and 24-h fasting, respectively, and the glucokinase-dependent glucosensing mechanism in the liver was identified as well as the significant activation of phospho-AMPK. However, fasting for 24 h did not activate glucosensing mechanisms and AMPK signaling pathways in the hypothalamus. On the other hand, significant reductions were observed in the mRNA levels of the lipid synthesis-related genes fas and accα, and the serum triglyceride levels as well. The mRNA levels of npy, agrp, pomc, and cart in the hypothalamus fluctuated during the fasting period without significant differences. With in situ hybridization npy signals upregulated in the ventral zone of posterior periventricular nucleus after 24-h fasting, pomc signals enhanced in the lateral tuberal nucleus. Based on the serum metabolomic analysis, the levels of branched-chain amino acids, butyrate, and short-chain acylcarnitine decreased, while those of medium- and long-chain acylcarnitine increased. Conclusion: Fasting for 24 h resulted in changes in npy and pomc signals within the hypothalamus and triggered the glucosensing mechanism in the liver of tilapia. This study is beneficial for elucidating the response of neuropeptides in the CNS to the changes of nutritional factors when hungry.

Effects of hazelnut soluble dietary fiber on lipid-lowering and gut microbiota in high-fat-diet-fed rats

Hazelnut is one of the most popular nuts in the world, rich in nutrients and various active substances. In this study, soluble dietary fiber (SDF) was extracted from hazelnut kernels, and its physicochemical properties and absorbability were explored. Hazelnut-SDF exhibited ideal water-holding, oil-holding and swelling capacity, and glucose, cholesterol and cholate absorbing ability. Scanning electron microscopy and fourier transform infrared spectroscopy showed that hazelnut-SDF had typical polysaccharide structure of functional groups. The main monosaccharides were identified as arabinose, rhamnose, xylose, ribose, glucuronic acid, mannose and glucose by gas chromatography–mass spectrometry. In high-fat diet rats, hazelnut-SDF could improve serum lipid parameters, inhibit lipid accumulation in liver and adipocytes, and regulate the expression level of liver lipid synthesis-related genes. It also could adjust intestinal short chain fatty acids, promote the composition and structure of intestinal microbiota, and significantly balance the abundance of Alloprevotella, Fusicatenibacter, Lactobacillus, Roseburia, Ruminococcaceae_UCG-005, Ruminococcaceae_UCG-014 and Clostridiales. The results concluded that oral administration of hazelnut-SDF could alleviate hyperlipidemia and obesity, and might serve as a potential functional food ingredient.

Sichuan Dark Tea-Based Medicated Dietary Formula Improves Obesity-Induced Renal Lipid Metabolism Disorder in Mice by Remodeling Gut Microbiota and Short-Chain Fatty Acid Metabolism

To investigate the renoprotective effects of a Sichuan dark tea-based medicated dietary formula (alternatively referred to as Qing, or clarity in Chinese) on mice with diet-induced obesity (DIO) and to explore the specific mechanisms involved. Male C57BL/6 mice were randomly assigned to three groups, a control group, a DIO group, and a Qing treatment group, or the Qing group, with 8 mice in each group. The mice in the control group were given normal maintenance feed and purified water, and the other two groups were fed a high-fat diet for 12 weeks to establish the DIO model. After that, high-fat diet continued in the DIO group, while the Qing group was given Qing at the same time for 12 weeks, during which period the weight of the mice was monitored and recorded every week. The mice were sacrificed after 12 weeks. Serum samples were collected and the levels of triglyceride (TG), total cholesterol (TC), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and albumin were measured to evaluate liver function. In addition, renal lipids were extracted to determine the levels of TG and TC in the kidney and periodic acid-Schiff (PAS) and oil red O stainings were performed to evaluate kidney pathological injury. Western blot was performed to determine the phosphorylated AMPK (pAMPK)/AMPK ratio in the kidney tissue. RT-qPCR and Western blot were used to determine the expression of proteins related to fatty acid oxidation, including acetyl-CoA carboxylase 1 (ACC1), carnitine acyltransferase 1 (CTP1), peroxisome proliferators-activated receptor γ (PPARγ), peroxisome proliferators-activated receptor-1 α (PPAR1α), sterol-regulatory element binding proteins (SREBP-1), and key proteins related to lipid synthesis, including fatty acid synthase (FASN) and stearoyl-coenzyme A desaturase 1 (stearoyl-CoA desaturase) in the kidney tissue. 16SrRNA and metabolomics were applied to analyze the gut microbiota in the intestinal contents and its metabolites. Compared with those of the control group, the levels of liver mass (P=0.0003), serum ALT (P<0.0001) and AST (P=0.0001), and kidney TC (P=0.0191) and TG (P=0.0101) of the DIO group were significantly increased and there was lipid deposition in the kidney. Compared with those of the DIO group, mice in the Qing group showed effective reduction in liver mass (P=0.0316) and improvements in the abnormal serum levels of AST (P=0.0012) and ALT (P=0.0027) and kidney TC (P=0.0200) and TG (P=0.0499). In addition, mice in the Qing group showed significant improvement in lipid deposition in the kidney. Qing group showed increased pAMPK/AMPK ratio in comparison with that of the DIO group. In comparison with those of the control group, mice in the DIO group had upregulated expression of lipid synthesis-related genes and proteins (SREBP-1, FASN, and SCD1). As for the fatty acid oxidation-related genes and proteins, DIO mice showed upregulated expression of ACC1 and downregulated expression of CPT1A, PPARγ, and PGC1α in comparison with those of the control group. In the Qing goup, improvements in regard to all these changes were observed. The Qing group demonstrated improvement in the disrupted homeostasis of the gut microbiota. Short-chain fatty acids in the cecal contents, especially isovaleric acid and propionic acid, were also restored. Sichuan dark tea-based medicated dietary formula may improve renal lipid metabolism by regulating gut microbiota and the levels of intestinal short-chain fatty acids, thereby protecting obesity-related kidney injury. Isovaleric acid and propionic acid may be the metabolites key to its regulation of gut microbiota.

β-Sitosterol Suppresses Lipopolysaccharide-Induced Inflammation and Lipogenesis Disorder in Bovine Mammary Epithelial Cells.

β-sitosterol, a natural plant steroid, has been shown to promote anti-inflammatory and antioxidant activities in the body. In this study, β-sitosterol was used to protect against lipopolysaccharide (LPS)-induced cell damage in bovine mammary epithelial cells, which are commonly studied as a cell model of mammary inflammatory response and lipogenesis. Results showed that treatment with a combination of LPS and β-sitosterol significantly reduced oxidative stress and inflammation, while increasing the expression of anti-apoptotic proteins and activating the hypoxia-inducible factor-1(HIF-1α)/mammalian target of rapamycin(mTOR) signaling pathway to inhibit apoptosis and improve lipid synthesis-related gene expression. Our finding suggests that β-sitosterol has the potential to alleviate inflammation in the mammary gland.

Dietary n-3/n-6 polyunsaturated fatty acid ratio modulates growth performance in spotted seabass (Lateolabrax maculatus) through regulating lipid metabolism, hepatic antioxidant capacity and intestinal health.

An 8-week feeding experiment was carried out to explore the effects of dietary n-3/n-6 polyunsaturated fatty acid (PUFA) ratio on growth performance, lipid metabolism, hepatic antioxidant status, and gut flora of spotted seabass (Lateolabrax maculatus). Six experimental diets were formulated to contain different levels of two purified oil sources including docosahexaenoic and eicosapentaenoic acids enriched oil (n-3) and linoleic acid-enriched oil (n-6) leading to n-3/n-6 PUFA ratios of 0.04, 0.35, 0.66, 1.35, 2.45 and 16.17. Each diet was fed to triplicate groups of juvenile L.maculatus (11.06±0.20g, 30 fish/tank). Final body weight (FBW), weight gain (WG), specific growth rates (SGR), protein efficiency ratio (PER) and feed utilization efficiency increased as n-3/n-6 PUFA ratio increased up to a certain level, and then decreased thereafter. Fish fed the diet with n-3/n-6 PUFA ratio of 0.66 exhibited the highest FBW, WG, SGR and PER and the lowest feed conversion ratio. Lower n-3/n-6 PUFA ratios induced up-regulated expression of lipid synthesis-related genes (fas, acc2 and srebp-1c) and down-regulated expression of lipolysis related genes (atgl, pparα, cpt-1 and aox). Higher expression of lipolysis-related genes (atgl, pparα and cpt-1) was recorded at moderate n-3/n-6 PUFA ratios (0.66 to 1.35). Moreover, inappropriate n-3/n-6 PUFA ratios triggered up-regulation of pro-inflammatory genes (il-6 and tnf-α) and down-regulation of anti-inflammatory genes (il-4 and il-10) in the intestine. The diet with n-3/n-6 PUFA ratio of 0.66 inhibited intestine inflammation, improved intestinal flora richness, increased the abundance of beneficial bacteria such as Lactobacillus, Alloprevotella and Ruminococcus, and reduced the abundance of harmful bacteria including Escherichia-Shigella and Enterococcus. In summary, it could be suggested that a dietary n-3/n-6 PUFA ratio of 0.66 can improve growth performance and feed utilization in L.maculatus, as is deemed to be mediated through regulation of lipid metabolism and intestinal flora.

High dietary energy decreased reproductive performance through increasing lipid deposition in Yangzhou geese at late laying stage

Dietary metabolizable energy (ME) level could offer a well production performance through maintaining lipid homeostasis in poultry. In this study, a total of 540 geese (450 females and 90 males) at 64 wk of age with similar body weight (4,600 ± 382) were randomly divided into 5 groups with 3 replicates in each group and 30 females and 6 males (1♂:5♀) in each replicate. After 2 wk adaptation, the 5 groups were designed to provide diet with ME intakes of 9.65, 10.05, 10.70, 11.45, and 11.75 MJ/kg, respectively, according to production requirement. Body weight, egg production, hatchability, blood lipid, and fat deposition were recorded after 6 wk feeding. The expression of lipid synthesis-related genes, lipoprotein lipase (LPL) and fatty acid synthase (FASN), were determined by quantitative real-time PCR. Geese fed with high ME diet of 11.75 MJ/kg caused an increased liver and abdominal fat weight and low hatchability of set eggs. The ovarian weight and oviduct length were higher in geese fed dietary energy of 10.7 MJ/kg as compared to the 9.65 MJ/kg groups, whereas no significant difference was observed in geese fed dietary energy of 10.05 MJ/kg. Dietary energy level did not change the concentration of serum lipids at the late egg laying stage. The LPL expression exhibited linear and quadratic effect in response to dietary ME. The FASN expression showed quadratic effect and a relatively higher expression was exhibited in 10.05 and 11.45 MJ/kg than that of the 9.65 and 10.70 MJ/kg ME groups. According to the productivity, reproductive performance, and fat deposition, dietary ME of 10.13 to 10.28 MJ/kg could be suggested for breeding geese at their late laying stage.

Lipid nanoparticle delivery of siRNA targeting Cyp2e1 gene attenuates subacute alcoholic liver injury in mice.

To investigate the effect and mechanism of lipid nanoparticle (LNP) delivery of small interfering RNA (siRNA) targeting Cyp2e1 gene on subacute alcoholic liver injury in mice. siRNA targeting Cyp2e1 gene was encapsulated in LNP (si-Cyp2e1 LNP) by microfluidic technique and the resulting LNPs were characterized. The optimal dose of si-Cyp2e1 LNP administration was screened. Forty female C57BL/6N mice were randomly divided into blank control group, model control group, si-Cyp2e1 LNP group, LNP control group and metadoxine group. The subacute alcoholic liver injury mouse model was induced by ethanol feeding for 10 d plus ethanol gavage for the last 3 d. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, and the superoxide dismutase (SOD) activity as well as malondialdehyde, reactive oxygen species, glutathione, triacylglycerol, total cholesterol contents in liver tissue were measured in each group, and liver index was calculated. The expression of genes related to oxidative stress, lipid synthesis and inflammation in each group of mice were measured by realtime RT-PCR. Compared with the model control group, the levels of liver index, serum ALT, AST activities, malondialdehyde, reactive oxygen species, triacylglycerol, total cholesterol contents in liver tissue decreased, but the SOD activity as well as glutathione increased in the si-Cyp2e1 LNP group (all P<0.01). Hematoxylin-eosin staining result showed disorganized hepatocytes with sparse cytoplasm and a large number of fat vacuoles and necrosis in the model control group, while the si-Cyp2e1 LNP group had uniformly sized and arranged hepatocytes with normal liver tissue morphology and structure. Oil red O staining result showed si-Cyp2e1 LNP group had lower fat content of the liver compared to the model control group (P<0.01), and no fat droplets accumulated. Anti-F4/80 monoclonal antibody fluorescence immunohistochemistry showed that the si-Cyp2e1 LNP group had lower cumulative optical density values compared to the model control group (P<0.01) and no significant inflammatory reaction. Compared with the model control group, the expression of catalytic genes P47phox, P67phox and Gp91phox were reduced (all P<0.01), while the expression of the antioxidant enzyme genes Sod1, Gsh-rd and Gsh-px were increased (all P<0.01). The mRNA expression of the lipid metabolism genes Pgc-1α and Cpt1 were increased (all P<0.01) and the lipid synthesis-related genes Srebp1c, Acc and Fasn were decreased (all P<0.01); the expression of liver inflammation-related genes Tgf-β, Tnf-α and Il-6 were decreased (all P<0.01). The si-Cyp2e1 LNP may attenuate subacute alcoholic liver injury in mice mainly by reducing reactive oxygen levels, increasing antioxidant activity, blocking oxidative stress pathways and reducing ethanol-induced steatosis and inflammation.

MTOR Plays a Conserved Role in Regulation of Nutritional Metabolism in Bivalve Sinonovacula constricta

The mammalian target of rapamycin (mTOR) has been shown to play a central role in regulating cell growth and metabolism. However, little is known about the function of mTOR in nutrient metabolism in bivalve mollusks. In this study, the role of mTOR in the regulation of nutrient metabolism was investigated in Sinonovacula constricta. First, the activation of mTOR was assayed after starvation and refeeding. Afterwards, the role of mTOR in the regulation of nutrient metabolism was investigated using an activator (MHY1485) or inhibitor (rapamycin) of mTOR. The open reading frame of the S. constricta mTOR is 7416 bp in length and encodes a polypeptide consisting of 2471 amino acids. The mTOR amino acid sequence of S. constricta was highly conserved when compared with other species and had a close evolutionary relationship with the TOR proteins of Crassostrea gigas and Lingula anatine. mTOR was expressed in the intestine, exhalent siphon, labial palppus, muscle, inhalent siphon, gill, mantle, digestive land, and gonad tissue of S. constricta, with the highest expression in muscle. During starvation, the level of phosphorylated mTOR protein was relatively low, and the ratio of LC3II/LC3I protein and the AMPKα mRNA level significantly increased with the increase in starvation time. After feeding, the level of phosphorylated mTOR protein increased from 0.13 to 0.56, and the ratio of LC3II/I protein and AMPKα mRNA level decreased from 1.17 to 0.38. MHY1485 significantly increased the level of phosphorylated 4E-BP1 and significantly decreased the ratio of LC3II/I proteins. Furthermore, MHY1485 significantly increased the mRNA level of the glucose metabolism-related gene glucokinase (GK), significantly decreased the mRNA expression of the G6P gene, and significantly increased the mRNA expression of the lipid synthesis-related genes sterol-regulatory element-binding protein (SREBP) and stearoyl-CoA desaturase (SCD). Rapamycin significantly reduced the level of phosphorylated 4E-BP1 and the mRNA expression of mTOR, and the expression level of phosphorylated 4EBP1 decreased from 0.97 to 0.28. Meanwhile, it also significantly reduced the mRNA expression of glucose metabolism-related genes GK, pyruvate kinase (PK), glucose transporter 1 (GLUT1), and G6P, as well as lipid synthesis-related genes SCD and acetyl-CoA carboxylase (ACC). These results indicate a conserved role of mTOR in regulating nutritional metabolism, including glucose metabolism, lipid synthesis, and autophagy in S. constricta.

Addition of solid-state fermentation product of yeast ameliorated the effects of high-fat diet on hepatic lipid metabolism, epidermal mucus, intestine and liver health, and gut microbiota of zebrafish

To assessed the effects of solid-state fermentation product of yeast (SFPY) supplementation on high-fat diet (HFD) induced challenges on growth, hepatic lipid metabolism, epidermal mucus, intestine and liver health and gut microbiota in zebrafish (Danio rerio), four experimental diets were prepared for one-month-old zebrafish: basal diet (Control), high-fat diet (HFD), 0.5% SFPY (0.5 SFPY) or 1.0% SFPY (1.0 SFPY)-added HFD. After 3 weeks of feeding, the results illustrated that although HFD increased weight gain (WG), corresponding decreased the feed conversation ratio (FCR), it caused negative influences including liver steatosis, intestinal and liver damage in zebrafish. Compared with HFD group, 1.0% SFPY obviously promoted the lysozyme activity and complement 4 (C4) level on epidermal mucus (P < 0.05), and significantly reduced liver TAG (P < 0.05). Addition of SFPY significantly decrease the lipid synthesis-related genes including C/EBPα, FAS, PPARγ, ACC1, DGAT2 and pro-inflammatory factors including IL-6, IL-1β in liver (P < 0.05), significantly increased the height of gut villi (P < 0.05). Moreover, the 1.0 SFPY group significantly reduced the abundance of gut microbiota concluding Proteobacteria, Fusobacteriota, Planctomycetota and improved the abundance of gut Actinobacteriota and Firmicutes (P < 0.05). To sum up, the addition of SFPY significantly ameliorated hepatic inflammation and intestinal damage caused by high-fat diet, and positively affected the intestinal microbiota composition of zebrafish.

Perilla, sunflower, and tea seed oils as potential dietary supplements with anti-obesity effects by modulating the gut microbiota composition in mice fed a high-fat diet.

Obesity has become a serious public health problem with its alarmingly increasing prevalence worldwide, prompting researchers to create and develop several anti-obesity drugs. Here, we aimed to investigate the protective effects of perilla seed oil (PSO), sunflower oil (SFO), and tea seed oil (TSO) against obesity through the modulation of the gut microbiota composition and related metabolic changes in mice fed a high-fat diet (HFD). Mice were divided into six equal groups: ND (normal diet); HFD; ORL (HFD supplemented with 20mg/kg body weight of orlistat); PSO, SFO, and TSO (HFD supplemented with 2g/kg body weight of PSO, SFO, and TSO, respectively). Our findings showed that PSO, SFO, and TSO supplementation significantly reduced body weight, organ weight, blood glucose, lipopolysaccharides (LPS), insulin resistance, and improved serum lipid levels (TG, TC, LDL-C, and HDL-C). Meanwhile, the three treatments alleviated oxidative stress and hepatic steatosis and reduced liver lipid accumulation. Relative mRNA expression levels of inflammatory cytokines (TNF-α, IL-1β, IL-6, and MCP-1) and lipid synthesis-related genes (PPAR-γ, FAS, and SREBP-1) were down-regulated, while β-oxidation-related genes (PPAR-α, CPT1a, and CPT1b) were up-regulated in the liver tissue of treated mice. Besides, dietary oil supplementation alleviated HFD-induced gut microbiota dysbiosis by promoting gut microbiota richness and diversity, decreasing the Firmicutes-to-Bacteroidetes ratio, and boosting the abundance of some healthy bacteria, like Akkermansia. PSO, SFO, and TSO supplementation could alleviate inflammation, oxidative stress, and hepatic steatosis, likely by modulating the gut microbiota composition in HFD-fed mice.

Structural characteristics of mixed pectin from ginseng berry and its anti-obesity effects by regulating the intestinal flora

Ginseng berry is the mature berry of ginseng and its polysaccharide has hypolipidaemic effect, but its mechanism remains unclear. A pectin (GBPA) with a molecular weight of 3.53 × 104 Da was isolated from ginseng berry, it was mainly composed of Rha (25.54 %), GalA (34.21 %), Gal (14.09 %) and Ara (16.25 %). Structural analysis showed that GBPA is a mixed pectin containing rhamnogalacturonan-I and homogalacturonan domains and has a triple helix structure. GBPA distinctly improved lipid disorders in obese rats, and changed intestinal flora with enrichments of Akkermansia, Bifidobacterium, Bacteroides and Prevotella, improved the levels of acetic acid, propionic acid, butyric acid and valeric acid. Serum metabolites which involved in the lipid regulation-related pathway, including cinnzeylanine, 10-Hydroxy-8-nor-2-fenchanone glucoside, armillaribin, 24-Propylcholestan-3-ol, were also greatly changed after GBPA treatment. GBPA activated AMP-activated protein kinase, phosphorylated acetyl-CoA carboxylase, and reduced the expression of lipid synthesis-related genes sterol regulatory element-binding protein-1c and fatty acid synthases. The regulatory effects of GBPA on lipid disorders in obese rats are related to the regulation of intestinal flora and activation of AMP-activated protein kinase pathway. Ginseng berry pectin could be considered in the future as a health food or medicine to prevent obesity.

A comparative study of dietary lipid sources on growth performance, body composition, fillet quality, and nutrient retention in juvenile yellow drum (Nibea albiflora)

Vegetable oils (VOs) are promising alternatives for fish oil (FO) in the aquafeed industry. However, high dietary VOs lead to undesirable effects on fish growth performance and health, but little information is known about VOs on fillet quality and nutrient retention. This study compared the impacts of different VOs on fish growth performance, fillet quality, and nutrient retention in juvenile yellow drums (Nibea albiflora). The FO group was used as the control (containing 6% FO), and the other four diets had the same lipid content with 5% FO was substituted by different VOs(soybean oil [SO], linseed oil [LO] rapeseed oil [RO], and palm oil [PO]). Juvenile fish (initial body weight of 5.55 ± 0.02 g) were fed the respective diets for 56 days. Results indicated the replacement of FO by VOs was feasible without affecting the growth performance, body composition, or serum biochemicals. The SO diet significantly increased fillet yield and fillet calcium (Ca) and phosphorus (P) element contents. The PO diet reduced water-holding capacity, and altered fillet texture, but significantly increased fillet n-3 LC-PUFAs retention. This could be attributed to the preferential oxidation of SFAs therefore selectively retained DHA in the fillet. Fillet transcriptome sequencing analysis revealed that compared with the FO group, the upregulated mRNA levels of lipid transport-related genes were found in the SO (fabp4, and cd36) and PO (fabp4, cd36, lcat, acbp, and cav1)groups, whereas lipid synthesis-related (acss, elovl4, acsbg, acsl, and elovl1) and lipolysis-related genes (mgll, hsl, acaa1, abhd5, decr1, acad9, pparα, and cpt1) expression were upregulated in the RO and PO groups, respectively. Weighted gene co-expression network analysis clustered two coexpression modules. One module was significantly and positively correlated with fillet yield (P = 0.01, R = 0.64) and Ca element retention (P = 0.05, R = 0.51), and contained genes related to fillet development (eno3 and tnnt3) and Ca regulation (dhrs7c, mcam, and ocam). The other module included genes related to lipid metabolism (elovl1, scd, tecr, fbn1, thbs1, and mdh1), which was associated with fillet n-3 LC-PUFAs retention. Collectively, these findings demonstrated that SO, LO, RO, and PO can be utilized as alternative lipid sources for FO. Based on transcriptome analysis results, genes related to fillet development, Ca regulation, and lipid metabolism were highlighted. These findings help to understand the relationship between dietary lipid sources and fillet nutrient retention in marine carnivorous fish species.