大鼠脂肪细胞分化过程中脂代谢相关基因转录表达时序的研究

对大鼠附睾和肾周脂肪组织分离的前体脂肪细胞原代培养，采用RT—PCR法分析脂代谢重要酶和转录因子基因在不同分化阶段转录表达的时序变化。结果显示，在前体脂肪细胞阶段，固醇调控元件结合蛋白（SREBP）-1c、碳水化合物反应元件结合蛋白（ChREBP）以及脂肪酸合成酶（FAS）、乙酰辅酶A羧化酶α（ACC1）、硬酯酰辅酶A去饱和酶（SCD）和激素敏感脂酶（HSL）基因均不转录表达；HSL mRNA在分化初期表达，中期表达水平最高，分化末期降低；SCD mRNA在分化中期表达，末期表达水平显著提高；分化初期FAS mRNA水平明显高于中期和末期；ACC1 mRNA仅在末期表达；分化初期SREBP-1c mRNA水平较高，中期和末期显著下降；ChREBP mRNA表达水平在分化末期稍高于中期，但差异不显著。表明，SREBP-1c mRNA的表达与脂肪细胞早期分化调控有关，分化成熟脂肪细胞的ChREBP mRNA表达可能与生脂基因的转录激活有关。     

Early development of porcine parthenogenetic embryos and reduced expression of primed pluripotent marker genes under the effect of lysophosphatidic acid

To further promote the early development of porcine embryos and capture “naïve” pluripotent state within blastocyst, the experiment explored the effects of lysophosphatidic acid (LPA) on the early development of porcine parthenogenetic embryos and the expression of pluripotency relevant genes. The results showed that the addition of 50 μM LPA significantly improved parthenogenetic embryo cleavage rate (82.7% vs. 74.7%, p < 0.05), blastocyst rate (24.5% vs. 11.3%, p < 0.05) and blastocyst cell count (56 ± 7.9 vs. 42 ± 1.0, p < 0.05) than that of the control group. In addition, immunostaining experiment determined that the fluorescence intensity of OCT4 was also significantly higher than that of the control group. The quantitative real‐time polymerase chain reaction (qRT–PCR) test revealed that addition of 50 μM LPA could significantly enhance the expression level of pluripotent gene OCT4 and trophoblast marker genes CDX2, however, decrease the expression of primitive hypoblast marker gene GATA4. The results also indicated that LPA might decrease the expression of GATA4 through the ROCK signalling pathway. For further investigating the effect of the addition of LPA on the expression of “primed” and “naïve” genes, we also detected the expression of those pluripotency‐related genes by qRT–PCR. The results showed addition of LPA had no significant effect on the expression of “naïve” pluripotent genes, but it was able to significantly decrease the expression of “primed” pluripotent genes, NODAL and Activin‐A; furthermore, it also could significantly improve the expression of OCT4 and c‐Myc which act as two important ES cell renewal factors. Above all, the addition of LPA can facilitate the early development of porcine parthenogenetic embryos, which may be able to benefit for capturing “naïve” pluripotency in vitro through inhibiting “primed” pluripotency.     

Effect of maternal rearing on lipid metabolism-related gene expression in offspring broilers during embryonic development

1. The aim of this study was to investigate the effect of maternal rearing on lipid metabolism and lipid metabolism-related gene expression in offspring broilers during embryonic development. 2. One hundred laying Sanhuang breeders were divided into two groups, and either floor-reared or cage-reared on the same diet. Liver and serum samples were extracted on days 14 and 19 of embryonic development and at hatching. The lipid metabolism related gene expressions of acetyl CoA carboxylase (ACC), fatty acid synthase (FAS), malic enzyme (ME), apolipoprotein B100(apoB100), sterol regulating element binding protein (SREBP-1c), carnitine palmitoyltransferase (CPT-1) and peroxisome proliferators-activated receptor (PPARα) genes were determined using real time RT-PCR. 3. The results showed that embryonic weight, liver weight, serum and hepatic total cholesterol (TC) concentration and serum triglyceride (TG) content were not significantly different between the cage-reared group and the floor-reared group during embryonic development. However, embryonic weight, liver weight, serum and hepatic TC concentration and serum TG content in the cage-reared group were significantly higher than in the floor-reared group at hatching. 4. Hepatic ACC, FAS, SREBP-1c, ME and apoB genes expression were not significantly different between the cage-reared and the floor-reared groups during E9 and E14 development. Hepatic ME gene expression in the cage-reared group was higher than in the floor-reared group during E19 development. However, hepatic FAS, SREBP-1c, CPT-1 and PPARα gene expressions in the cage-reared group was higher than in the floor-reared group. 5. A change in the maternal regime could regulate lipid metabolism in offspring broilers during embryonic development, and especially at hatching.     

Effect of dietary fish oil on fatty acid deposition and expression of cholesterol homeostasis controlling genes in the liver and plasma lipid profile: comparison of two animal models

The objective of the present study was to compare hepatic fatty acid deposition, plasma lipid level and expression of cholesterol homeostasis controlling genes in the liver of rats (Wistar Albino; n = 32) and pigs (Large White × Landrace; n = 32) randomly assigned into two groups of 16 animals each and fed 10 weeks the diet with either 2.5% of fish oil (F; source of eicosapentaenoic and docosahexaenoic acid, EPA+DHA) or 2.5% of palm oil (P; high content of saturated fatty acids; control). F‐rats deposited in the liver three times less EPA, but 1.3 times more DHA than F‐pigs (p < 0.05). Dietary fish oil relative to palm oil increased PPARα and SREBP‐2 gene expression much strongly (p < 0.01) in the pig liver in comparison with the rat liver, but expression of Insig‐1 and Hmgcr genes in the liver of the F‐pigs relative to the expression of these genes in the liver of the P‐pigs was substantially lower (p < 0.01 and p < 0.05 respectively) as compared to rats. When plasma lipid concentration in the F‐animals was expressed as a ratio of the plasma concentration in the P‐counterparts, dietary fish oil decreased HDL cholesterol less (p < 0.01), but LDL cholesterol and triacylglycerols more (p < 0.05 and p < 0.001 respectively) in rats than in pigs: more favourable effect of fish oil on rat plasma lipids in comparison with pigs can therefore be concluded. Concentration of total cholesterol and both its fractions in the rat plasma was negatively correlated (p < 0.01) with hepatic DHA, but also with unsaturated myristic and palmitic acid respectively. It has been concluded that regarding the similarity of the plasma lipid levels to humans, porcine model can be considered superior; however, using this model, dietary fish oil at the tested amount (2.5%) was not able to improve plasma lipid markers in comparison with saturated palm oil.     

Effect of dietary fish oil on the expression of genes involved in lipid metabolism in liver and skeletal muscle of lactating sows

This study investigated the hypothesis that dietary supplementation of fish oil as a source of n‐3 polyunsaturated fatty acids (PUFA) influences the expression of target genes of sterol regulatory element‐binding proteins (SREBP)‐1 and (SREBP)‐2 involved in triacylglycerol (TAG) synthesis and fatty acid and cholesterol metabolism in the liver, and moreover activates the expression of target genes of peroxisome proliferation‐activated receptor (PPAR)‐α involved in TAG and fatty acid catabolism in liver and skeletal muscle. Twenty lactating sows were fed a control diet or a fish oil diet with either 50 g of a mixture of palm oil and soya bean oil (4:1, w/w) or fish oil per kg. The diet of the fish oil group contained 19.1 g of n‐3 PUFA (mainly 20:5 n‐3 and 22:6 n‐3) per 100 g of total fatty acids, while the diet of the control group contained 2.4 g of n‐3 PUFA (mainly 18:3 n‐3) per 100 g of total fatty acids. The fish oil group had reduced relative mRNA concentrations of various target genes of SREBP‐1 involved in fatty acid and TAG synthesis in comparison with the control group (p < 0.05). Relative mRNA concentrations of target genes of PPARα involved in fatty acid catabolism in both liver and muscle, and mRNA concentrations of target genes of SREBP‐2 involved in cholesterol synthesis and uptake were not influenced by fish oil supplementation. Concentrations of cholesterol and TAG in plasma, fat content of milk and weight gains of litters during the suckling period were not different between the two groups of sows. In conclusion, this study suggests that fish oil has only minor effects on hepatic lipid metabolism, which are non‐critical with respect to milk production in sows.     

催乳素对奶牛脂肪细胞脂代谢关键基因的作用

催乳素(prolactin,PRL)是启动和维持泌乳的重要激素,在泌乳期间能调节机体将营养物质和能量优先流向乳腺,用于乳脂和乳蛋白的合成。本试验采用体外分离培养奶牛前脂肪细胞,诱导分化为成熟脂肪细胞,加入PRL探讨其对脂肪细胞脂代谢相关基因的作用。结果显示,奶牛白色脂肪细胞存在催乳素受体(prolactin receptor,PRLR),且在PRL质量浓度为75μg/L,作用时间为9h时,PRLR表达量达到最高,此时调节脂合成的核转录因子SREBPE-1c及下游控制脂合成的基因FAS和LPL均显著降低,75μg/L组与对照组相比差异极显著(P0.01);控制脂分解的基因HSL显著升高,75μg/L组与对照组相比差异显著(P0.05);细胞培养上清液中的甘油含量显著升高,75μg/L组与对照组相比差异显著(P0.05)。结果表明,PRL能直接作用于奶牛白色脂肪细胞,降低脂合成作用并促进脂分解作用。     

The effect of a combined dietary treatment with cholesterol and cholic acid on the lipid metabolism of geese at low or high choline concentrations.

A previous study demonstrated that a dietary treatment of young geese with cholesterol and cholic acid raises lipid concentrations in the liver. The present study was carried out to investigate whether such a lipid accumulation caused by those hyperlipidemic compounds can be intensified by low dietary choline concentrations. Therefore, 38 eight-week old geese were divided into four groups of 9 or 10 animals each and received a basal diet poor in choline which consisted predominately of maize and soy protein isolate over a period of 8 weeks. Treatment factors were supplementation of diets with cholesterol and cholic acid (0 vs. 5 g of cholesterol and cholic acid each per kg) and supplementation of choline chloride (0 vs. 1.5 g/kg). Final body weights as well as carcass weights were neither influenced significantly by dietary treatment with cholesterol and cholic acid nor by low dietary choline concentrations. However, feeding diets supplemented with cholesterol and cholic acid markedly increased liver weights (two-fold), hepatic triglyceride (3.7-fold) and cholesterol (12-fold) concentrations and percentages of monounsaturated fatty acids at the expense of saturated and polyunsaturated fatty acids in the liver. In geese fed diets with cholesterol and cholic acid, insufficient choline supply did not intensify, but even slightly reduced hepatic lipid accumulation. Geese fed diets with cholesterol and cholic acid exhibited markedly increased levels of cholesterol, triglycerides and phospholipids in plasma and very low-density lipoproteins, regardless of the choline supply. Muscle tissue of geese fed diets supplemented with cholesterol and cholic acid exhibited also increased concentrations of triglycerides and cholesterol whereas the fatty acid composition of muscle lipids remained unchanged. Among geese without hyperlipidemic treatment, concentrations of triglycerides in plasma and very low-density lipoproteins as well as the concentrations of phosphatidylcholine in liver and muscle tissue were not reduced by low dietary choline concentrations. Therefore, it is suggested that those animals were able to synthesize endogenous sufficient choline.     

Effect of reduced dietary protein and supplementation with a docosahexaenoic acid product on broiler performance and meat quality

1. Chicken breast meat is a lean meat due to its low content of intramuscular fat (IMF) resulting in an overall lower acceptability by consumers due to a decrease in juiciness, flavour and increased chewiness. Recently, studies performed in pigs suggested the possibility of increasing IMF by decreasing dietary crude protein (CP) content, an effect possibly mediated through an increased lipogenesis.

2. Dietary supplementation with lipids rich in omega 3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA) may modulate an increase in the content of these fatty acids in meat from monogastric animals and, thus, promote the daily intake of n-3 LC-PUFA by humans.

3. LC-PUFA are very susceptible to oxidation, resulting in off-flavours that affect meat quality and consumers’ acceptability.

4. This trial was conducted to assess the effect of reducing dietary CP, from 21% to 17%, on chicken’s meat IMF content and, simultaneously, to evaluate if a complementary supplementation with a proprietary n-3 LC-PUFA source (DHA Gold?) could improve meat quality. These effects were assessed by measuring productive performance and meat quality, oxidative stability, sensory traits and fatty acid profile.

5. A reduction in CP content of broiler diets, from 21% to 17%, balanced for lysine, improved performance while it was not sufficient to increase IMF content in chicken meat. In contrast, DHA Gold? supplementation had a positive impact both in broiler productive parameters and in meat fatty acid profile.

6. In addition, incorporation of 7.4% of DHA Gold? in the diet promoted carcass yield but negatively affected chicken meat acceptability by consumers, due to a decrease of meat oxidative stability.

7. Overall the data suggest that neither a dietary supplementation with DHA Gold? nor a reduction in CP have a direct positive effect in the levels of IMF present in broiler meat.     

The effect of dietary protein levels on the expression of genes coding for four selected protein translation initiation factors in muscle tissue of Wujin pig

The objective of this study was to investigate the regulatory mechanism underlying the increased muscle protein accumulation in pigs while were fed a high protein diet. The eukaryotic initiation factors (eIFs) have been reported to involve in muscle protein synthesis. We investigated the mRNA and protein expression levels of eIF2B1, 4A1, 4B and 4E in Wujin pigs fed either a high protein (HP: 18%) or a low protein (LP: 14%) diet at 30, 60 or 100 kg body weight, based on real‐time PCR and western blotting analyses. Our results indicated that the expression levels of eIF2B1 mRNA and protein were increased by HP diet at all body weight. The HP diet showed higher mRNA and protein levels of eIF4B gene at 60 and 100 kg. The protein expression of eIF4E phosphorylation was increased by HP diet only at 30 kg. These data suggested that the HP diet promoted porcine muscle protein accumulation mainly by up‐regulating eIF2B1, 4B and 4E rather than 4A1 expression along the growth stages.     

Fortification of pork loins with docosahexaenoic acid (DHA) and its effect on flavour

Pork is traditionally low in docosahexanoic acid (DHA, C22:6n-3) and deficient in omega-3 fats for a balanced human diet. DHA as triglycerides was commercially prepared from the microalgae Schizochytrium and injected into fresh pork loins. Treatments of a mixed brine control (CON), 3.1% sunflower oil in mixed brine (SF) and a 3.1% DHA oil in mixed brine (DHA) were injected into pork loins at 10 mL/100 g and grilled at 205°C. After cooking, the CON and SF pork loins contained 0.03 to 0.05 mg DHA/g of pork and the DHA injected loins contained approximately 1.46 mg DHA/g. This also changed the fatty acid profile of omega-6: omega-3 from, 5 to 1 in the CON pork, to a ratio of 1.7 to 1 in DHA pork. The appearance, odor, oxidation rates and sensory taste, as judged by a trained panel, determined the DHA injected meat to be, ''slightly desirable’ and gave lower ''off flavour’ scores, relative to the CON and SF injected pork. Pork can be fortified with DHA oil to 146 mg/100 g serving, which would meet half the recommended daily omega 3 fatty acid requirements for adult humans and would be desirable in taste.     

Fortification of pork loins with docosahexaenoic acid(DHA) and its effect on flavour

Pork is traditionally low in docosahexanoic acid(DHA, C22:6n-3) and deficient in omega-3 fats for a balanced human diet. DHA as triglycerides was commercially prepared from the microalgae Schizochytrium and injected into fresh pork loins. Treatments of a mixed brine control(CON), 3.1% sunflower oil in mixed brine(SF) and a 3.1% DHA oil in mixed brine(DHA) were injected into pork loins at 10 mL/100 g and grilled at 205°C. After cooking, the CON and SF pork loins contained 0.03 to 0.05 mg DHA/g of pork and the DHA injected loins contained approximately1.46 mg DHA/g. This also changed the fatty acid profile of omega-6: omega-3 from, 5 to 1 in the CON pork, to a ratio of 1.7 to 1 in DHA pork. The appearance, odor, oxidation rates and sensory taste, as judged by a trained panel,determined the DHA injected meat to be, ‘slightly desirable' and gave lower ‘off flavour' scores, relative to the CON and SF injected pork. Pork can be fortified with DHA oil to 146 mg/100 g serving, which would meet half the recommended daily omega 3 fatty acid requirements for adult humans and would be desirable in taste.     

Effects of dietary copper on the expression of lipogenic genes and metabolic hormones in steers

An experiment was conducted to determine the effects of Cu supplementation on performance, subcutaneous adipose tissue mRNA expression of acetyl CoA carboxylase (ACC), stearoyl CoA desaturase (SCD), uncoupling protein 2 (UCP2), and leptin in growing and finishing steers. Forty-eight purebred Angus steers were allotted to one of five treatments: 1) control (no supplemental Cu); 2) 10 mg Cu/kg DM from CuSO4; 3) 10 mg Cu/kg DM from a Cu amino acid complex (Availa Cu); 4) 20 mg Cu/kg DM from CuSO4; 5) 20 mg Cu/kg DM from Availa Cu. Steers were fed an alfalfa hay corn-based diet for 56 d (basal diet contained 7.1 mg Cu/kg DM) and switched to a high-concentrate diet for 144 d (basal diet contained 6.1 mg Cu/kg DM). Blood samples were obtained every 28 d throughout the entire experiment. On d 112 of the finishing period, subcutaneous adipose tissue biopsies were obtained from the tailhead of three animals per treatment and analyzed for ACC, SCD, UCP2, and leptin mRNA expression. Animal performance was not affected by Cu supplementation during the growing phase. Steers receiving 10 mg Cu/kg DM from Availa Cu had higher (P < 0.05) ending body weights and tended (P < 0.10) to have higher ADG than steers receiving 10 mg Cu/kg DM from CuSO4 during the finishing phase. Serum concentrations of nonesterified fatty acid and insulin were not affected by Cu supplementation. Steers receiving supplemental Cu tended (P < 0.11) to have less backfat relative to controls. However, dietary Cu did not influence the level of subcutaneous adipose tissue ACC and SCD mRNA. Neither UCP2 nor leptin gene expression was affected by Cu supplementation. These results indicate that dietary Cu supplementation (10 to 20 mg Cu/kg DM diet) may alter lipid metabolism of subcutaneous adipose tissue; however, it does not seem to affect expression of certain lipogenic genes.     

Effects of dietary phytol on tissue accumulation of phytanic acid and pristanic acid and on the tissue lipid profiles in mice

Recent in vitro evidence suggests that the phytol‐derived fatty acids, phytanic acid (PA) and pristanic acid (PrA), are components of animal products with the potential to cause both beneficial and harmful effects on human health. In this study, we investigated the in vivo tissue accumulation of PA and PrA and the changes in tissue lipid profiles, using mice fed a phytol‐containing diet. After 4 weeks of treatment with a diet containing 1.0% phytol, plasma, adipose tissue, liver, and brain were collected and their lipid profiles were biochemically and gas‐chromatographically determined. Dietary phytol caused PA and PrA accumulation in the adipose tissue and liver but not in the brain, and reduced plasma and liver triacylglycerol levels. Phytol intake also decreased the fatty acid concentrations in the adipose tissue, especially polyunsaturated fatty acids such as linoleic acid, but increased the concentrations of these fatty acids in the liver. However, dietary phytol had a low impact on the brain lipid profile. This study suggests that dietary phytol intake caused accumulation of PA and PrA and modified lipid profiles in the adipose tissue and liver, but that the brain is an insusceptible tissue to dietary phytol‐induced changes.     

The effect of maternal obesity on fatty acid transporter expression and lipid metabolism in the full‐term placenta of lean breed swine

This study was conducted to evaluate the influence of back‐fat thickness (BF), at mating of sows, on the maternal and newborn circulating lipids, expression of placental fatty acids (FA) transporters and lipid accumulation in placenta. Full‐term placentas were obtained by vaginal delivery from BFI (9–14 mm; n = 37), BFII (15–19 mm; n = 43) and BFIII (20–27 mm; n = 38) sows according to BF at mating, and frozen placental sections were analysed for fat accumulation. Blood samples were collected from the sows of day 105 pregnancy and from cord blood at delivery. mRNA and protein expression levels were evaluated with real‐time RT‐PCR and Western blotting. Our results demonstrated that BFII females had significantly increased litter weight and placental efficiency, decreased maternal triglyceride (TG) and non‐esterified fatty acids (NEFA) levels, decreased maternal IL‐6, TNFα and leptin levels compared to BFIII females (p < .05). BFIII sows were associated with significantly decreased newborn TG levels, increased newborn glucose, IL‐6 and TNFα levels compared to BFI or BFII sows (p < .05). BFI and BFII females had significantly decreased placental TG, NEFA and cholesterol (CHOL) contents compared to BFIII females (p < .05). Moreover, decreased CD36, FATP1, FABP4, and FABP1 mRNA and protein and FATP4 protein expression, and increased LPL activity were also observed in BFIII group compared with BFII group (p < .05). PPARγ mRNA and protein and lipogenic genes such as SREBP‐1c, ACSL1, ACCα, FAS and SCD mRNA expression were downregulated or upregulated, respectively, in the placentas of BFIII sows compared to BFI or BFII sows (p < .05). Overall, this study demonstrated that there is no advantage, in terms of litter live size, litter weight and placental FA transport and metabolism, in performing the mating of sows with BF>19 mm.