Effects of long-chain fatty acids on cytosolic triacylglycerol accumulation and lipid droplet formation in primary cultured bovine mammary epithelial cells

Mammary epithelial cells have recently been shown to express and secrete leptin into milk and to accumulate triacylglycerol (TAG) in cytosol. We examined the effects on the accumulation of cytosolic TAG of free fatty acid addition to the medium bathing bovine mammary epithelial cells (bMEC). Both saturated (palmitic and stearic) and unsaturated (oleic and linoleic) fatty acids stimulated the accumulation of TAG in a concentration-dependent manner from 50 to 400 microM and the expression of mRNA expression for CD36, which is involved in the uptake and secretion of long-chain fatty acids. However, leptin mRNA expression and lipid droplet formation were significantly increased only by the addition of unsaturated, but not saturated, fatty acids. Interestingly, both types of fatty acids stimulated alphas1-casein mRNA expression. These data suggest that the expression of leptin is related to droplet formation, whereas CD36 is related to cytosolic TAG accumulation, and that fatty acids or cytosolic TAG accumulation also have a role to accelerate differentiation of bMEC as shown by casein synthesis.     

Lipid quality and filleting yield of reared meagre (Argyrosomus regius)

Reared meagre (Argyrosomus regius) of average weight 1278 g was evaluated for its filleting yield and fillet lipid quality. Filleting yield averaged 42.2%, while very low levels of muscle fat deposits were measured (1.06%). Analysis of lipid classes revealed a high proportion of polar lipids (48.3–59.1%) and low levels of neutral fatty acids (40.9–51.7%) when compared to corresponding values of other farmed fish species. Phosphatidyl‐choline (PC) was the dominant polar lipid (2.92 mg g^?1 muscle), while cholesterol the most abundant neutral lipid (1.68 mg g^?1 muscle). The fatty acid composition of meagre fillet generally reflects the dietary fatty acids. Among n‐3 fatty acids, eicosapentaenoic (4.58%) and docohexaenoic (15.0%) were measured to be the most abundant ones, and 18:2n‐6 (11.9%) was the most common n‐6 fatty acid. The fatty acid profiles of polar and neutral fractions differ, with monounsaturated fatty acids being predominant in the neutral and n‐3 polyunsaturates in the polar lipids. The high polar lipid contents and n‐3 fatty acids and the low atherogenic and thrombogenic indexes indicate a high quality of this species’ lipids.     

褐点石斑鱼鱼肉及其副产物的脂肪酸分析

为了评估褐点石斑鱼的脂肪酸组成特点,采用固相萃取和气相色谱技术对鱼肉、头、皮和内脏的脂质含量、总脂及各脂类脂肪酸组成,甘油三酯(triacylglycerols, TAG)和磷脂(phospholipids, PL)中脂肪酸的位置分布进行了较全面的分析。石斑鱼鱼头和内脏中含有丰富的糖脂(saccharolipids, SL)和PL。在各组织总脂中,必需脂肪酸的含量占总脂肪酸的24.17%～24.51%,二十二碳六烯酸(docosahexaenoic acid, DHA)及二十碳五烯酸(eicosapentaenoic acid, EPA)的含量为12.42%～13.73%,动脉粥状硬化指数、血栓形成指数和n-6/n-3的比例处于较低水平。多不饱和脂肪酸(polyunsaturated fatty acids, PUFA),尤其是DHA+EPA(28.43%～37.37%)在PL中的比例要远高于在其它脂类中的占比。饱和脂肪酸主要分布在TAG和PL的sn-2位,单不饱和脂肪酸主要分布在TAG的sn-1,3位和PL的sn-2位,PUFA倾向于分布在TAG的sn-1,3位和PL的sn-1位;T...     

Variation of lipid class composition in Nitzschia laevis as a response to growth temperature change

The lipid composition and the distribution of fatty acids in the lipid pool were determined in eicosapentaenoic acid (EPA)-producing microalga (Nitzschia laevis) grown under different temperatures. Both the relative amounts of lipid classes and the degree of fatty acid unsaturation in various lipid species were not greatly changed under tested growth conditions. Higher temperature up to 23 °C benefited the growth of N. laevis but only had a slight influence on EPA and lipid contents. Further increasing the culture temperature caused a serious inhibition of both the cell growth and fatty acid biosynthesis. Under all temperatures tested, triacylglycerol (TAG) was the predominant lipid constituent (64.5–69.1% of total lipid) and was highly saturated. Lower temperature favored the formation of polar lipids. The highest content of phosphatidylcholine (PC), the major phospholipids component, was reached at 15 °C (10.9% of total lipid). In sharp contrast to TAG, PC was highly unsaturated and contained a higher amount of EPA under lower temperature. The highest EPA content in polar lipid was achieved at 19 °C. The results from this investigation suggested that the low temperature could improve the distribution of polyunsaturated fatty acids in phospholipids, though it could not significantly influence their amount, especially in PC.     

Composition of meat and offal from weaned and fattened rabbits and results of stereospecific analysis of triacylglycerols and phosphatidylcholines

BACKGROUND: Rabbit meat has excellent nutritive properties. The purpose of this study was to characterize rabbit meat and offal; in particular, the lipid fraction was studied in order to evaluate total and positional fatty acid (FA) compositions of triacylglycerol (TAG) and phosphatidylcholine (PC) fractions. Eight samples of weaned and eight of fattened rabbits were considered. RESULTS: Fattened rabbit meat contained slightly higher protein percentage content (P < 0.05) in comparison to weaned (20.1% versus 18.0%). Calcium content was higher in meat than in offal, unlike sodium, iron, zinc, manganese and copper. The cholesterol content in offal was much higher than in meat. FA profiles of total lipid showed a high percentage of unsaturated fatty acids and an n‐6/n‐3 ratio of 10.3 for fattened rabbit meat. Stereospecific analysis of TAG and PC was carried out on an eight‐sample pool of each meat and offal from weaned and fattened rabbits. In all samples the sn‐2‐position was prevalently esterified with oleic and linoleic acids in TAG, with polyunsaturated fatty acids in PC. CONCLUSION: Lipids from rabbit meat presented higher content of monounsaturated FA and lower n‐6/n‐3 ratio in comparison to offal, which was characterized by higher cholesterol and mineral levels. Copyright © 2011 Society of Chemical Industry     

胆碱对脂肪代谢调控及其机制的研究进展

胆碱作为动物生长发育必须的营养物质,广泛存在于动物产品及植物种子中,在动物体内主要以卵磷脂、磷酸胆碱、乙酰胆碱等形式存在。近年来,通过建立磷脂酰乙醇胺N-甲基转移酶(phosphatidylethanolamine N-methyltransferase,PEMT)模型的研究表明,胆碱对机体脂肪代谢作用十分明显。日粮中适量添加胆碱可有效降低动物肝脏脂肪沉积从而缓解脂肪肝的发生,对提高肉品质及抗氧化能力有积极作用。着重阐述了胆碱对肝脏和肌肉等组织脂肪代谢的影响,说明胆碱对肉品质及与胰岛素抵抗的关系,分析探讨其调控机制。     

Role of diacylglycerol O-acyltransferase (DGAT) isoforms in bovine hepatic fatty acid metabolism

Fatty acid accumulation in hepatocytes induced by high concentrations of fatty acids due to lipolysis and the associated oxidative damage they cause occur most frequently after calving. Because of their role in esterification of fatty acids, diacylglycerol acyltransferase isoforms (DGAT1 and DGAT2) could play a role in the susceptibility of dairy cows to develop fatty liver. To gain mechanistic insights, we performed in vivo and in vitro analyses using liver biopsies or isolated primary hepatocytes. The in vivo study (n = 5 cows/group) involved healthy cows [average liver triacylglycerol (TAG) = 0.78%; 0.58 to 0.93%, ratio of triglyceride weight to wet liver weight] or cows diagnosed with fatty liver (average TAG = 7.60%; 5.31 to 10.54%). In vitro, hepatocytes isolated from 3 healthy female calves (1 d old, 44 to 53 kg) were challenged with (fatty acids) or without (control) a 1.2 mM mixture of fatty acids in an attempt to induce metabolic stress. Furthermore, hepatocytes were treated with DGAT1 inhibitor or DGAT2 inhibitor for 2 h followed by a challenge with (DGAT1 inhibitor + fatty acids or DGAT2 inhibitor + fatty acids) or without (DGAT1 inhibitor or DGAT2 inhibitor) the 1.2 mM mixture of fatty acids for 12 h. Data analysis of liver biopsies was compared using a 2-tailed unpaired Student's t-test. Data from calf hepatocyte treatment comparisons were assessed by one-way ANOVA, and multiplicity for each experiment was adjusted by the Holm's procedure. Data indicated that both fatty liver and in vitro challenge with fatty acids were associated with greater mRNA and protein abundance of SREBF1, FASN, DGAT1, and DGAT2. In contrast, mRNA and protein abundance of CPT1A and very low-density lipoprotein synthesis-related proteins MTTP and APOB were markedly lower. However, compared with fatty acid challenge alone, DGAT1 inhibitor + fatty acids led to greater mRNA and protein abundance of CPT1A and APOB, and greater mRNA abundance of SREBF1 and MTTP. Furthermore, this treatment led to lower mRNA abundance of FASN and DGAT2 and TAG concentrations. Compared with fatty acid challenge alone, DGAT2 inhibitor + fatty acids led to greater mRNA and protein abundance of CPT1A, MTTP, and APOB, and lower mRNA and protein abundance of SREBF1 and FASN. In addition, compared with control and fatty acids, there was greater protein abundance of GRP78 and PERK in both DGAT1 and DGAT2 inhibitor with or without fatty acids. Furthermore, compared with control and fatty acids, reactive oxygen species concentrations in the DGAT1 inhibitor with or without fatty acid group was greater. Overall, data suggested that DGAT1 is particularly relevant in the context of hepatocyte TAG synthesis from exogenous fatty acids. Disruption of both DGAT1 and DGAT2 altered lipid homeostasis, channeling fatty acids toward oxidation and generation of reactive oxygen species. Both DGAT isoforms play a role in promoting fatty acid storage into TAG and lipid droplets to protect hepatocytes from oxidative damage.     

TOCOPHEROLS AND PHOSPHOLIPIDS ENHANCE THE OXIDATIVE STABILITY OF BORAGE AND EVENING PRIMROSE TRIACYLGLYCEROLS

The effect of individual phospholipids and tocopherols as well as their combinations on the oxidative stability of borage and evening primrose triacylglycerols (TAG) was investigated. Based on conjugated dienes (CD) and 2-thiobarbituric acid reactive substances (TBARS) formed in borage and evening primrose TAG, the antioxidant effectiveness of phosphatidylcholine (PC), was higher than that of phosphotidylethanolamine (PE) in borage TAG while the reverse was observed for evening primrose TAG. Meanwhile, a synergistic effect between tocopherols and phospholipids in both borage and evening primrose TAG was noted. The most effective combination in borage TAG was that of PC and α-tocopherol, while PE with α-tocopherol exerted the best effects in evening primrose TAG. Therefore, the antioxidant effectiveness of phospholipids may vary depending on their fatty acid composition, nature of the functional groups and the chemical composition of the lipid system in which they are examined. The results also demonstrate the importance of minor components of vegetable oils on their oxidative stability.     

Compositional characteristics of muscle and visceral oil from steelhead trout and their oxidative stability

The quality of lipid from steelhead trout viscera, a byproduct of steelhead trout industry, was evaluated and compared with that of the muscle. Steelhead trout viscera had a higher lipid content than muscle and the visceral oil differed from muscle oil in its lipid class composition. Neutral lipids, mainly triacylglycerols (TAG), comprised the major lipid class in both muscle and visceral lipid. However, as expected, muscle contained a remarkably higher level of phospholipids (PL) than viscera, and the ratio of total neutral lipids to polar lipids was lower in muscle than in viscera. Visceral and muscle lipid had similar fatty acid compositions, with the concentration of muscle polyunsaturated fatty acids (PUFA) slightly higher than that of visceral PUFA. Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were the major n-3 PUFA present in both muscle and visceral lipid; docosapentaenoic acid (DPA) was present at about 1.61–1.76%. With respect to minor components, muscle lipid had higher α-tocopherol and total carotenoids contents than visceral lipid. Oxidative stability of visceral and muscle lipid, as determined by conjugated dienes (CD) and thiobarbituric acid reactive substances (TBARS), showed that visceral lipid was more stable against oxidation than muscle lipid, probably due to their different lipid profiles. Besides, carotenoids, which were present more abundantly in muscle, might have acted as prooxidants and therefore, decreased the oxidative stability of muscle lipid. These results suggest that steelhead trout viscera may serve as a good source of lipid and could be utilized for production of omega-3 oils.     

Fluorescence resonance energy transfer analysis of mitochondrial:lipid association in the porcine oocyte

The role of endogenous lipid in the provision of energy during in vitro maturation of immature porcine oocytes has been studied. Fluorescence resonance energy transfer (FRET) acceptor bleaching methods have been used to examine mitochondrial:lipid droplet co-localisation in live oocytes. FRET experiments demonstrate whether organelles are within the FRET-distance (i.e. 6-10 nm), thus showing true association on a molecular scale. Immature and in vitro-matured porcine oocytes were stained with Mitotracker Green (MTG; mitochondria) and Nile Red (NR; lipid droplets). The data indicated sufficient overlap between MTG emission and NR excitation to support a FRET reaction and that mitochondria and lipid droplets were sufficiently co-localised for a FRET reaction to occur. When NR-stained lipid droplets were specifically bleached, a significant increase in the MTG signal in stained mitochondria was observed (FRET efficiency, E=22.2 +/- 3.18%). These results strongly suggest a metabolic role for lipid metabolism during oocyte maturation. This conclusion was reinforced by the use of inhibitors of fatty acid beta-oxidation, methyl palmoxirate or mercaptoacetate, exposure to which during oocyte maturation led to developmental failure post-fertilisation. These data provide strong evidence that MTG and NR can act as a FRET pair and that in porcine oocytes, mitochondria and lipid droplets lie within 6-10 nm of each other, indicating association on a molecular scale. The findings also suggest that endogenous triglycerides play an important role in energy metabolism during porcine in vitro maturation.     

Microwave heating influences on fatty acid distributions of triacylglycerols and phospholipids in hypocotyl of soybeans(glycine max L.)

Soybeans were exposed to microwave heating for 6, 12 or 20 min at a frequency of 2450 MHz. The hypocotyls were then separated from the other tissues, and the lipid components and the positional distribution of fatty acids in triacylglycerols (TAG) and phospholipids (PL) were investigated. Major lipid components were TAG and PL, while steryl esters (SE), free fatty acids (FFA), and 1,3- and 1,2-diacylglycerols (DAG) were minor ones. Following microwave heating, a significant increase (p<0.05) was observed in FFA and in both forms of DAG (primarily 1,3-DAG). The greatest rate of PL losses (p<0.05) was observed in phosphatidylethanolamine (PE), followed by phosphatidylcholine (PC) and phosphatidylinositol (PI). Significant differences (p<0.05) in fatty acid distributions occurred (with few exceptions) when soybeans were microwaved for 12 min or more. Nevertheless, the principal characteristics for the positional distribution of fatty acids still remained after 20 min of microwave heating.     

冻融循环对熟制鲣鱼暗色肉脂质变化的影响

为探究冻融循环对熟制鲣鱼暗色肉脂质变化的影响,对熟制鲣鱼暗色肉进行冻融循环（0、2、4、6、8 次）贮藏,测定冻融过程中样品的过氧化值（peroxide value,POV）、酸价（acid value,AV）、pH值、脂质组成、脂肪酸组成等指标。结果显示,在冻融循环贮藏过程中,脂质组成发生明显变化,甘油三酯（triacylglycerol,TAG）和磷脂（polar lipid,PL）质量分数明显降低,游离脂肪酸（free fatty acid,FFA）质量分数明显升高。在冻融循环过程中暗色肉的pH值显著减小,POV和AV显著增加（P＜0.05）。脂质中总饱和脂肪酸和总多不饱和脂肪酸含量都逐渐下降,TAG和PL中脂肪酸含量明显下降,其中PL中总多不饱和脂肪酸含量下降明显,FFA中总多不饱和脂肪酸含量显著升高（P＜0.05）。主成分分析（principal component analysis,PCA）和偏最小二乘判别分析（partial least squares discriminant analysis,PLS-DA）结果表明,经过不同冻融次数的鱼肉之间存在一定的差异性,且随着冻融次数的增加脂质氧化程度不断加剧。相关性分析结果表明FFA质量分数与POV、AV呈正相关,与PL质量分数、TAG质量分数、pH值呈负相关。实验结果表明冻融循环会影响熟制鲣鱼暗色肉脂质组成,同时引起脂质的氧化分解。这为熟制鲣鱼暗色肉在运输和贮藏过程中脂质控制提供一定的理论指导。     

LIPID AND LIPID SOLUBLE COMPONENTS OF GONADS OF GREEN SEA URCHIN (STRONGYLOCENTROTUS DROEBACHIENSIS)

Seasonal changes in the lipid and lipid soluble components of gonads of green sea urchin (Strongylocentrotus droebachiensis) were investigated. The lipid content was highest in the spring while it was lowest in the winter. The composition of nonpolar and polar lipid classes of sea urchin gonads were not affected qualitatively, but there were significant (P<0.05) quantitative variations. Major nonpolar lipid classes were triacylglycerols (TAG), free fatty acids (FFA) and sterols (ST) while dominant polar lipid components were phosphatidylcholine (PC) and phosphatidylethanolamine (PE). The fatty acids 16:0 and 14:0 were the major saturated fatty acids (SFA) consistently present in total, nonpolar and polar lipids of sea urchin gonads. The 5-olefinic acid 20:1 n-15 was the dominant monounsaturated fatty acid (MUFA) while 20:4 n-6 and 20:5 n-3 were the major polyunsaturated fatty acids (PUFA) present. The content of PUFA increased during cold climatic conditions with a simultaneous decrease in the content of SFA. These changes were reversed during warm seasons. Total carotenoid content was highest in the spring. Echininone was the major carotenoid present and its relative content varied significantly (P<0.05) with the season; being highest in the spring and lowest in the summer. In addition, β-carotene was present in each season. Thus, the importance of seasonal variations on the lipid and lipid soluble components of sea urchin gonads is evident.     

小麦胚芽贮藏期内脂质水解酸败机理解析

以小麦胚芽为研究对象,构建脂质水解酸败的模拟体系以及原位体系,解析小麦胚芽贮藏期内脂质水解酸败的机理。结果表明:光照、氧气和脂肪氧合酶未对小麦胚芽模拟体系中的游离脂肪酸含量产生明显影响,而脂肪酶则显著提升游离脂肪酸生成量;此外,游离脂肪酸生成量随水分活度增加而增加,于水分活度为0.75时达峰值,且小麦胚芽内部结合水和不易流动水含量也呈显著增加趋势,水分以及脂质流动性增加;小麦胚芽原位体系中游离脂肪酸含量随贮藏时间延长而逐渐增加,磷脂酰胆碱含量恰相反,且小麦胚芽脂质体膜遭到破坏,游离出的甘油三酯聚集形成油滴。由此可见,小麦胚芽脂质水解酸败模式主要表现为其脂质体膜中的磷脂酰胆碱降解而释放甘油三酯,而后甘油三酯经由脂肪酶催化水解生成大量的游离脂肪酸,从而产生水解酸败现象。     

Identification of 14 new single nucleotide polymorphisms in the bovine SLC27A1 gene and evaluation of their association with milk fat content

The solute carrier family 27 member 1 (SLC27A1) is an integral membrane protein involved in the transport of long-chain fatty acids across the plasma membrane. This protein has been implicated in diet-induced obesity and is thought to be important in the control of energy homeostasis. In previous reports, our group described the isolation and characterization of the bovine SLC27A1 gene. The bovine gene is organized in 13 exons spanning over more than 40 kb of genomic DNA and maps in BTA 7 where several quantitative trait loci for fat related traits have been described. Because of its key role in lipid metabolism and its genomic localization, in the present work the search for variability in the bovine SLC27A1 gene was carried out with the aim of evaluating its potential association with milk fat content in dairy cattle. By sequencing analysis of all exons and flanking regions 14 new single nucleotide polymorphisms (SNPs) were identified: 1 in the promoter, 7 in introns and 6 in exons. Allele frequencies of all the SNPs were calculated by minisequencing analysis in two groups of Holstein-Friesian animals with highest and lowest milk-fat content estimated breeding values as well as in animals of two Spanish cattle breeds, Asturiana de los Valles and Menorquina. In the conditions assayed, no significant differences between Holstein-Friesian groups were found for any of the SNPs, suggesting that the SLC27A1 gene may have a poor or null effect on milk fat content. In Asturiana and Menorquina breeds all the positions were polymorphic with the exception of SNPs 1 and 8 in which C allele was fixed in both of them.     

Distribution of fatty acids in triacylglycerols and phospholipids from peas (Pisum sativum L.)

BACKGROUND: The fatty acid distribution of triacylglycerols (TAG) and major phospholipids (PL) obtained from four varieties of peas (Pisum sativum) was investigated. The total lipids extracted from the peas were separated by thin layer chromatography into seven fractions. RESULTS: The major lipid components were PL (52.2–61.3%) and TAG (31.2–40.3%), while hydrocarbons, steryl esters, free fatty acids and diacylglycerols (sn‐1,3 and sn‐1,2) were also present in minor proportions (5.6–9.2%). The main PL components isolated from the four varieties were phosphatidylcholine (42.3–49.2%), phosphatidylinositol (23.3–25.2%) and phosphatidylethanolamine (17.7–20.5%). Significant differences (P < 0.05) in fatty acid distribution were found for different pea varieties. Phosphatidylinositol was unique in that it had the highest saturated fatty acid content among the three PL. However, the principal characteristics of the fatty acid distribution in the TAG and three PL were evident among the four varieties: unsaturated fatty acids were predominantly located in the sn‐2 position while saturated fatty acids primarily occupied the sn‐1 or sn‐3 position in the oils of the peas. CONCLUSION: These results should be useful to both producers and consumers for the manufacture of pea foods in Japan. Copyright © 2007 Society of Chemical Industry     

肌肉细胞膜脂肪酸组成和变化对猪肉风味的影响

脂肪酸在猪肉风味的形成中扮演重要的角色,脂肪酸的氧化是影响猪肉风味的主要原因之一。细胞膜上磷脂部分脂肪酸的氧化,是肌肉系统在细胞内膜水平上的磷脂组分变化,它决定了肉风味的形成。因此,猪肉肌肉细胞膜脂肪酸对猪肉风味的形成有着重要意义。本文主要综述猪肉肌肉细胞膜脂肪酸的组成、代谢及其氧化和挥发对于猪肉风味的影响及其调控。