长链多不饱和脂肪酸EPA、DHA的基因工程研究进展

长链多不饱和脂肪酸(long chain polyunsaturated fatty acid,LCPUFA)对维持人体健康具有重要作用,对其需求逐年增加,但是由于环境污染与渔业资源的下滑,有限的鱼油资源越来越不能满足人们需求。运用现代生物技术人们已相继分离了多个LCPUFA合成相关基因,并阐明了多条LCPUFA合成代谢途径。通过转基因技术在高等植物中成功合成了对人体健康十分重要的长链多不饱和脂肪酸,尤其是二十碳五烯酸(EPA),二十二碳六烯酸(DHA)。综述了LCPUFA的合成途径及转基因研究的最新进展,分析合成LCPUFA存在的问题及解决方法,并对未来多不饱和脂肪酸EPA,DHA的基因工程研究进行展望。     

四个脂肪酸合成酶基因在哺乳动物细胞中的超表达提高长链多不饱和脂肪酸生物合成效率

DHA(22:6n-3)、EPA(20:5n-3)和ARA(20:4n-6)三种长链多不饱和脂肪酸在生物体内活性最强,它们在促进大脑发育和功能维持以及在预防和治疗心血管疾病、炎症、癌症等多种疾病方面有着重要作用。然而,尽管哺乳动物体内有完整的长链多不饱和脂肪酸合成酶系,但哺乳动物合成这些长链多不饱和脂肪酸的效率很低而主要依赖于食物获取。本研究应用转基因方法,将哺乳动物来源的Δ6和Δ5脂肪酸去饱和酶以及Δ6和Δ5脂肪酸延长酶这4种酶的编码基因构建成为一个多基因表达载体,然后转染哺乳动物细胞HEK293T,实现了4个目的基因的超表达,再通过气质联用(GC-MS)分析证实了DHA、EPA和ARA等长链多不饱和脂肪酸的合成效率及水平显著增加,DHA的水平更是提高了2.5倍。由此可见,哺乳动物具有某种抑制长链多不饱和脂肪酸高水平合成的机制,但通过Δ6和Δ5脂肪酸去饱和酶以及Δ6和Δ5脂肪酸延长酶的超表达,能够打破哺乳动物这种抑制机制,从而显著提高DHA、EPA、ARA等的合成水平。同时,本研究的思路也为在转基因动物中生产长链多不饱和脂肪酸提供了重要的启示。     

二十碳五烯酸和二十二碳六烯酸的生物合成途径研究进展

以二十碳五烯酸(EPA)和二十二碳六烯酸(DHA)为代表的长链多不饱和脂肪酸,对人体心血管系统、神经系统、抗炎免疫系统等有着理想的功效,因而倍受研究者关注。我们从结构、生物来源和合成、生理功能及生物工程途径的研究现状等方面阐述了EPA和DHA的相关信息,并对其生成途径中涉及的酶进行了简要概述。     

水生动物脂肪酸延长酶基因研究进展

多不饱和脂肪酸(polyunsaturated fatty acids,PUFAs)对人类具有重要的生理作用,但由于人类缺乏从头合成这些脂肪酸的能力,而必须从食物中获得,鱼类等水生动物中二十碳五烯酸(20:5n-3,EPA)和二十二碳六烯酸(22:6n-3,DHA)等多不饱和脂肪酸含量丰富,从而使得鱼类等水生动物成为人们获得PUFAs的主要途径。鱼类等水生动物体内PUFAs的合成是在一系列去饱和酶和延长酶的作用下实现的,其中长链脂肪酸延长酶是合成PUFAs必不可少的一类关键酶。本研究总结了水生动物长链多不饱和脂肪酸合成的过程,综述了水生动物中脂肪酸延长酶基因的研究进展和未来的发展方向。     

脂肪酸对离体培养中华绒螯蟹肝胰腺PUFAs合成相关基因表达的影响

长期以来海水水生动物和淡水水生动物多不饱和脂肪酸合成能力一直是众多研究的重点。相比于淡水水生动物,海水水生动物更容易从食物中获得多不饱和脂肪酸,因此其合成多不饱和脂肪酸的能力低于淡水水生动物。中华绒螯蟹作为一种洄游型甲壳动物,其脂肪酸合成能力的研究显得更加重要。本研究通过中华绒螯蟹肝胰腺离体培养技术,利用不同的脂肪酸培养中华绒螯蟹肝胰腺组织,旨在探讨不同脂肪酸对中华绒螯蟹肝胰腺多不饱和脂肪酸合成相关基因表达的影响,为研究饲料中添加不同脂肪酸对中华绒螯蟹多不饱和脂肪酸合成的影响提供了一定的理论参考依据。本研究表明:油酸会显著增加中华绒螯蟹肝胰腺ELOVL6、fad6、和fad9基因的表达量;不同脂肪酸对fad6和fad9基因表达量的影响相似,且相比于EPA和DHA,亚麻酸和油酸会显著增加fad6和fad9基因的表达。通过本研究,我们得出EPA和DHA等多不饱和脂肪酸会抑制多不饱和脂肪酸合成的过程。进一步证实了海水水生动物多不饱和脂肪酸合成能力和海水中高含量的多不饱和脂肪酸之间的关系。     

大豆ω-3和ω-6脂肪酸脱氢酶基因密码子优化

DHA和EPA对心脑血管疾病以及癌症等疾病的防治有着重要的作用,其中ω-3和ω-6脂肪酸脱氢酶是合成多不饱和脂肪酸DHA和EPA过程中的两个关键的去不饱和酶,由于缺乏这两种酶的基因,高等动物无法自发合成多不饱和脂肪酸,为了得到可以生产多不饱和脂肪酸的转基因动物,本研究计划将大豆的这两个去饱和脂肪酸作为目的基因,但由于不同物种之间的密码子使用具有偏爱性,会严重影响到目的基因在宿主体内的表达水平,为了解决这一问题,提高目的基因在宿主体内的表达水平,同时采用Jcat在线软件和手动替换密码子两种方法对大豆的ω-3和ω-6脂肪酸脱氢酶基因进行了优化,然后选出最优的优化方法,为后期进一步的实验提供目的基因。最后本文预测了两种优化结果的RNA二级结构和CAI、CBI、Nc值和GC含量,比较得出手动替换的结果优于在线优化的结果。     

真菌发酵生产鱼油特征脂肪酸

DHA和EPA是鱼油的特征脂肪酸,具有调整血脂、改善大脑功能和维持正常视力等重要作用.一直以来,其来源主要是海产鱼油.近年研究发现许多微生物的脂质中也含有DHA和EPA,特别是一些低等真菌含量甚为丰富,利用其发酵生产DHA和EPA替代鱼油资源具有广阔的前景.阐述了能产生EPA,DHA的真菌及其合成途径和发酵影响因素.     

ω-3多不饱和脂肪酸抗肿瘤新途径——抑制具核梭杆菌黏附宿主细胞

【背景】大量文献报道ω-3多不饱和脂肪酸尤其是二十二碳六烯酸(Docosahexaenoic Acid,DHA)与二十碳五烯酸(Eicosapentaenoic Acid,EPA)具有抗肿瘤作用,但是其抗肿瘤机制还不够完善。【目的】探究ω-3多不饱和脂肪酸、具核梭杆菌以及结直肠癌三者之间的关联。【方法】在检测二十二碳六烯酸、二十碳五烯酸、α-亚麻酸(α-Linolenic Acid,ALA)等ω-3多不饱和脂肪酸对人结直肠腺癌细胞Caco-2、正常结肠上皮细胞NCM460生长影响的基础上,检测DHA等3种多不饱和脂肪酸对具核梭杆菌黏附人体细胞以及Fap2、FadA、RadD等具核梭杆菌毒力关键基因表达的影响。【结果】30μg/mL的DHA、EPA、ALA对Caco-2生长抑制分别为9.09%、4.95%、7.52%,而对NCM460生长抑制达31.15%、25.48%、29.11%,而且相关抑制作用仅具有浓度依赖性而无时间依赖性。经30μg/mL的DHA、EPA、ALA预处理的具核梭杆菌黏附Caco-2细胞的能力分别下降81.04%(P=0)、93.63%(P=0)和68.63%(P=0);而共培养时加入DHA、EPA、ALA对具核梭杆菌黏附Caco-2细胞的能力没有显著影响。同时,30μg/mLDHA处理导致F.nucleatum的Fap2基因显著下降10.22%(P=0.027);30μg/mL EPA处理导致FadA、Fap2基因分别显著下降23.49%(P=0)、15.09%(P=0.003);30μg/mL ALA处理导致FadA基因显著下降26.75%(P=0.012)。【结论】综合上述实验结果以及DHA、EPA、ALA仅能短时间抑制具核梭杆菌生长等文献报道,我们认为,DHA、EPA等ω-3多不饱和脂肪酸并非简单地直接杀伤或抑制肿瘤细胞和F.nucleatum;抑制FadA、Fap2等黏附相关基因表达,降低F.nucleatum黏附宿主细胞能力是其抗肿瘤作用的关键组成部分。ω-3多不饱和脂肪酸等活性物质对F.nucleatum等在结直肠肿瘤发生、发展中发挥重要作用的肠道细菌的影响与机制应深入开展研究。     

转基因植物生产超长链多不饱和脂肪酸研究进展

超长链多不饱和脂肪酸(VLCPUFAs)对人类健康非常重要。日常摄入一定量的VLCPUFAs能够补充人体自身合成的不足, 并对某些疾病起到明显的预防和治疗作用。VLCPUFAs主要源自深海鱼油, 但由于市场需求的迅速增长和海洋可捕捞 鱼类资源的日益减少, 该途径已经远远不能满足市场的需要, 寻找更为持续且稳定的VLCPUFAs来源已经成为当务之急。最近, 人们已经克隆了VLCPUFAs生物合成相关的去饱和酶和延伸酶基因, 并希望在植物特别是油料作物中共表达这些基因, 使其成为生产VLCPUFAs的“绿色细胞工厂”。目前已有多个研究小组在进行转基因植物合成VLCPUFAs的探索, 并取得了突破性的研究成果。本文综述了相关的研究进展, 并对存在的问题和解决策略进行了探讨。     

Elovl5转基因果蝇产生更长链的脂肪酸

大多数昆虫体内的多不饱和脂肪酸 (Polyunsaturated fatty acids,PUFAs) 主要是碳链长为18个碳原子的多不饱和脂肪酸 (C18-PUFAs),几乎不含价值更高、生物活性更强的C20、C22等长链的多不饱和脂肪酸。文中利用黑腹果蝇Drosophila melanogaster (Fruit fly) 作为生物模型,将来源于小鼠的Δ6-脂肪酸延长酶Elovl5基因优化后转移到果蝇中使其表达。结果表明通过显微注射法成功将含有Elovl5基因的载体导入果蝇胚胎中,在荧光显微镜下检测到在果蝇整个发育阶段均有增强型绿色荧光蛋白 (Enhanced green fluorescent protein,EGFP) 表达,同时Elovl5基因的表达显著地促使果蝇体内C18多不饱和脂肪酸向着更长链多不饱和脂肪酸的生物合成方向转化。使用转基因技术得到体内富含C20、C22等长链多不饱和脂肪酸的转基因果蝇模型,将为进一步开展果蝇多不饱和脂肪酸生物合成的机制提供研究基础。     

Metabolism and functional effects of plant-derived omega-3 fatty acids in humans

Alpha-linolenic acid (ALA) is an essential fatty acid and the substrate for the synthesis of longer-chain, more unsaturated ω-3 fatty acids, eicosapentaenoic acid (EPA), docosapentaenoic acid and docosahexaenoic acid (DHA). EPA and DHA are associated with human health benefits. The primary source of EPA and DHA is seafood. There is a need for sustainable sources of biologically active ω-3 fatty acids. Certain plants contain high concentrations of ALA and stearidonic acid (SDA). Here we review the literature on the metabolism of ALA and SDA in humans, the impact of increased ALA and SDA consumption on concentrations of EPA and DHA in blood and cell lipid pools, and the extent to which ALA and SDA might have health benefits. Although it is generally considered that humans have limited capacity for conversion of ALA to EPA and DHA, sex differences in conversion to DHA have been identified. If conversion of ALA to EPA and DHA is limited, then ALA may have a smaller health benefit than EPA and DHA. SDA is more readily converted to EPA and appears to offer better potential for health improvement than ALA. However, conversion of both ALA and SDA to DHA is limited in most humans.     

Eicosapentaenoic acid and docosahexaenoic acid effects on tumour mitochondrial metabolism, acyl CoA metabolism and cell proliferation

In order to investigate the effects of high-fat diets rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), Wistar rats bearing subcutaneous implants of the Walker 256 tumour were fed pelleted chow containing low DHA/EPA or high DHA/EPA. The presence of n-3 polyunsaturated fatty acids (PUFAs) led to a marked suppression (35-46%) of tumour growth over a 12 day period. Both the whole tumour homogenate and the Percoll-purified mitochondrial fraction presented significant changes in fatty acid composition. The levels of EPA increased in both n-3 dietary groups while the levels of DHA increased only in the high DHA/EPA group, in comparison with the control chow-fed group. The presence of n-3 PUFAs led to an increase in mitochondrial acyl CoA synthetase activity, but neither the cytoplasmic acyl CoA content nor the n-3 fatty acid composition of the cytoplasmic acyl CoAs was altered by the diet. The content of thiobarbituric acid-reactive substances (TBARS) was increased in the low DHA/EPA group but was unchanged in the high DHA/EPA group. In vitro studies with the Walker 256 cell line showed a 46% decrease in cell growth in the presence of either EPA or DHA which was accompanied by a large decrease in the measured mitochondrial membrane potential. The TBARS content was increased only in the EPA-exposed cells. Cell cycle analysis identified a decrease in G0-G1 phase cells and an increase in G2-M phase cells and apoptotic cells, for both EPA and DHA-exposed cells. The data show that the presence of n-3 PUFAs in the diet is able to significantly after the growth rate of the Walker 256 tumour. The involvement of changes in mitochondrial membrane composition and membrane potential have been indicated for both EPA and DHA, while changes in lipid peroxidation have been identified in the presence of EPA but not of DHA.     

Omega-3 fatty acids in cellular membranes: a unified concept

The Omega-3 fatty acid DHA (docosahexaenoic acid, 22:6) and its sister molecule EPA (eicosapentaenoic acid, 20:5) are highlighted here. These highly unsaturated fatty acids are widespread in nature, especially in the marine environment, and are essential in membranes ranging from deep sea bacteria to human neurons. Studies of DHA/EPA in bacteria have led to a working model on the structural roles of these molecules and are described in this review. The main points are: (a) genomic analysis shows that genes encoding the DHA/EPA pathways are similar, supporting the idea that structural roles in bacteria might be similar, (b) biochemical analysis shows that DHA and EPA are produced in bacteria by a polyketide process distinct from the pathway of plants and animals; this allows DHA and EPA to be produced in anaerobic or oxygen-limited environments, (c) regulatory systems triggered by temperature and pressure have been identified and studied, and add to the understanding of the roles of these molecules, (d) DHA/EPA bacteria are located almost exclusively in the marine environment, raising the prospect of an important linkage between membrane processes and marine conditions, (e) physiological studies of an EPA recombinant of E. coli show that EPA phospholipids contribute essential fluidity to the bilayer and that an EPA-enriched membrane supports a respiratory lifestyle dependent on proton bioenergetics; the EPA recombinant displays other physiological properties likely attributed to high levels of EPA in the bilayer, and (f) chemical studies such as chemical dynamic modeling support the idea that DHA and presumably EPA contribute hyperfluidizing properties to the membrane. We hypothesize that DHA/EPA phospholipids contribute fluidity and other properties to the bilayer which distinguish these highly unsaturated chains from monounsaturates and polyunsaturates such as 18:2 and 18:3. We further hypothesize that the structural properties of DHA/EPA functioning in bacteria are also harnessed by higher organisms for enhancing crucial membrane processes including photosynthesis and energy transduction.     

Cardiovascular disease prevention and treatment

The incidence of fatal and non-fatal cardiovascular disease (sudden cardiac death (SCD), myocardial infarction, others) varies, depending on conventional risk factors. However, in Western countries, like the US or Germany, incidences of fatal and non-fatal cardiovascular disease are far higher than in countries like Japan. In the present article, these differences are discussed and related to eicosapentaenoic acid (C20:5omega-3 or C20:5n-3; EPA) and docosahexaenoic acid (C22:6omega-3; DHA). Dietary intake of EPA and DHA and a number of other factors determine levels of EPA and DHA in an individual—best assessed as the omega-3 index, defined as the percentage of EPA and DHA in red cells, and analyzed in a standardized fashion. A review of the literature, expanded by measurements of the omega-3 index, indicates that the risk of sudden cardiac death correlates inversely with the omega-3 index. For persons with an omega-3 index <4%, risk is tenfold, as compared to persons with an omega-3 index >8%. A similar, less-pronounced, correlation exists for non-fatal cardiovascular disease. EPA and DHA have anti-arrhythmic and anti-atherosclerotic mechanisms of action. In large-scale intervention studies, intake of EPA and DHA has been demonstrated to reduce SCD and non-fatal cardiovascular events. Assessing or recommending dietary intake of EPA and DHA does not predict the resulting omega-3 index. Taken together, the omega-3 index is a biomarker to assess a person's content of omega-3 fatty acids, and thus the risk for sudden cardiac death, as well as non-fatal cardiovascular events. EPA and DHA prevent fatal and non-fatal cardiovascular disease and complications of congestive heart failure.     

Long-Chain Omega-3 Polyunsaturated Fatty Acids Have Developmental Effects on the Crop Pest,the Cabbage White Butterfly Pieris rapae

Nutritional enhancement of crops using genetic engineering can potentially affect herbivorous pests. Recently, oilseed crops have been genetically engineered to produce the long-chain omega-3 polyunsaturated fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) at levels similar to that found in fish oil; to provide a more sustainable source of these compounds than is currently available from wild fish capture. We examined some of the growth and development impacts of adding EPA and DHA to an artificial diet of Pieris rapae, a common pest of Brassicaceae plants. We replaced 1% canola oil with EPA: DHA (11:7 ratio) in larval diets, and examined morphological traits and growth of larvae and ensuing adults across 5 dietary treatments. Diets containing increasing amounts of EPA and DHA did not affect developmental phenology, larval or pupal weight, food consumption, nor larval mortality. However, the addition of EPA and DHA in larval diets resulted in progressively heavier adults (F _{4, 108} = 6.78; p = 0.011), with smaller wings (p < 0.05) and a higher frequency of wing deformities (R = 0.988; p = 0.001). We conclude that the presence of EPA and DHA in diets of larval P. rapae may alter adult mass and wing morphology; therefore, further research on the environmental impacts of EPA and DHA production on terrestrial biota is advisable.     

Omega-3 fatty acids,EPA and DHA induce apoptosis and enhance drug sensitivity in multiple myeloma cells but not in normal peripheral mononuclear cells

The n-3 polyunsaturated fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have been shown to enhance the effect of chemotherapeutic drugs in clinical studies in cancer patients and to induce apoptotic tumor cell death in vitro. Until now, EPA and DHA have never been investigated in multiple myeloma (MM). Human myeloma cells (L363, OPM-1, OPM-2 and U266) and normal peripheral blood mononuclear cells were exposed to EPA and DHA, and effects on mitochondrial function and apoptosis, caspase-3 activation, gene expression and drug toxicity were measured. Exposure to EPA and DHA induced apoptosis and increased sensitivity to bortezomib in MM cells. Importantly, they did not affect viability of normal human peripheral mononuclear cells. Messenger RNA expression arrays showed that EPA and DHA modulated genes involved in multiple signaling pathways including nuclear factor (NF) κB, Notch, Hedgehog, oxidative stress and Wnt. EPA and DHA inhibited NFκB activity and induced apoptosis through mitochondrial perturbation and caspase-3 activation. Our study suggests that EPA and DHA induce selective cytotoxic effects in MM and increase sensitivity to bortezomib and calls for further exploration into a potential application of these n-3 polyunsaturated fatty acids in the therapy of MM.     

Differential effects of eicosapentaenoic acid and docosahexaenoic acid in promoting the differentiation of 3T3-L1 preadipocytes

The objective of this study was to determine the effects of enrichment with n-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), on the differentiation of 3T3-L1 preadipocytes. Enrichment with DHA but not EPA significantly increased the differentiation markers compared to control differentiated cells. DHA compared to EPA treatment led to a greater increase in adiponectin secretion and, conditioned media collected from DHA treated cells inhibited monocyte migration. Moreover, DHA treatment resulted in inhibition of pro-inflammatory signaling pathways. DHA treated cells predominantly accumulated DHA in phospholipids whereas EPA treatment led to accumulation of both EPA and its elongation product docosapentaenoic acid (DPA), an n-3 fatty acid. Of note, adding DPA to DHA inhibited DHA-induced differentiation. The differential effects of EPA and DHA on preadipocyte differentiation may be due, in part, to differences in their intracellular modification which could impact the type of n-3 fatty acids incorporated into the cells.     

Omega-3 fatty acids from fish oils and cardiovascular disease

Fish and fish oils contain the omega-3 fatty acids known as eicosapentaenoic acid (EPA) plus docosahexaenoic acid (DHA). Epidemiological studies have shown an inverse relation between the dietary consumption of fish containing EPA/DHA and mortality from coronary heart disease. These relationships have been substantiated from blood measures of omega-3 fatty acids including DHA as a physiological biomarker for omega-3 fatty acid status. Controlled intervention trials with fish oil supplements enriched in EPA/DHA have shown their potential to reduce mortality in post-myocardial infarction patients with a substantial reduction in the risk of sudden cardiac death. The cardioprotective effects of EPA/DHA are widespread, appear to act independently of blood cholesterol reduction, and are mediated by diverse mechanisms. Their overall effects include anti-arrhythmic, blood triglyceride-lowering, anti-thrombotic, anti-inflammatory, endothelial relaxation, plus others. Current dietary intakes of EPA/DHA in North America and elsewhere are well below those recommended by the American Heart Association for the management of patients with coronary heart disease. (Mol Cell Biochem 263: 217–225, 2004)     

Engineering of EPA/DHA omega-3 fatty acid production by Lactococcus lactis subsp. cremoris MG1363

Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have been shown to be of major importance in human health. Therefore, these essential polyunsaturated fatty acids have received considerable attention in both human and farm animal nutrition. Currently, fish and fish oils are the main dietary sources of EPA/DHA. To generate sustainable novel sources for EPA and DHA, the 35-kb EPA/DHA synthesis gene cluster was isolated from a marine bacterium, Shewanella baltica MAC1. To streamline the introduction of the genes into food-grade microorganisms such as lactic acid bacteria, unnecessary genes located upstream and downstream of the EPA/DHA gene cluster were deleted. Recombinant Escherichia coli harboring the 20-kb gene cluster produced 3.5- to 6.1-fold more EPA than those carrying the 35-kb DNA fragment coding for EPA/DHA synthesis. The 20-kb EPA/DHA gene cluster was cloned into a modified broad-host-range low copy number vector, pIL252m (4.7 kb, Ery) and expressed in Lactococcus lactis subsp. cremoris MG1363. Recombinant L. lactis produced DHA (1.35?±?0.5 mg g^?1 cell dry weight) and EPA (0.12?±?0.04 mg g^?1 cell dry weight). This is believed to be the first successful cloning and expression of EPA/DHA synthesis gene cluster in lactic acid bacteria. Our findings advance the future use of EPA/DHA-producing lactic acid bacteria in such applications as dairy starters, silage adjuncts, and animal feed supplements.     

Eicosapentaenoic and docosahexaenoic acid ethyl esters differentially enhance B-cell activity in murine obesity

EPA and DHA are not biologically equivalent; however, their individual activity on B cells is unknown. We previously reported fish oil enhanced murine B-cell activity in obesity. To distinguish between the effects of EPA and DHA, we studied the ethyl esters of EPA and DHA on murine B-cell function as a function of time. We first demonstrate that EPA and DHA maintained the obese phenotype, with no improvements in fat mass, adipose inflammatory cytokines, fasting insulin, or glucose clearance. We then tested the hypothesis that EPA and DHA would increase the frequency of splenic B cells. EPA and DHA differentially enhanced the frequency and/or percentage of select B-cell subsets, correlating with increased natural serum IgM and cecal IgA. We next determined the activities of EPA and DHA on ex vivo production of cytokines upon lipopolysaccharide stimulation of B cells. EPA and DHA, in a time-dependent manner, enhanced B-cell cytokines with DHA notably increasing IL-10. At the molecular level, EPA and DHA differentially enhanced the formation of ordered microdomains but had no effect on Toll-like receptor 4 mobility. Overall, the results establish differential effects of EPA and DHA in a time-dependent manner on B-cell activity in obesity, which has implications for future clinical studies.