多不饱和脂肪酸对细胞膜功能影响的研究进展

多不饱和脂肪酸是细胞膜磷脂的重要组成成份,影响细胞膜的稳定性.它具有广泛的生物学功能,包括细胞内信号传导通路、基因表达和细胞凋亡的调控等.主要从细胞膜脂质的组成,细胞膜的流动性及膜脂质过氧化等方面对多不饱和脂肪酸对细胞膜功能的影响进行了综述.     

对骨细胞生物学功能的最新认识

骨细胞长久以来都被看作是一种终末分化的、代谢隋性的、深埋在骨基质之中的占位细胞．并不具有重要的生理作用和功能。然而近些年的研究发现,骨细胞是一种活跃的多功能细胞,参与机体诸多生物学过程的调控。骨细胞的生物学功能可以大致归纳为以下六类：调节骨重建平衡、感知和转导力学刺激、参与机体的神经-内分泌调节、与肌肉组织有密切的相互作用、降解与合成骨基质及调控骨组织内储存的钙和其他生物活性物质。     

三维回转骨细胞条件培养基对成骨细胞功能的调节作用

骨细胞是骨组织中主要的力学感受器.研究失重条件下骨细胞对效应细胞的调控作用对于揭示失重引起的骨丢失机制具有重要意义.本研究拟采用三维回转器模拟失重,探讨模拟失重骨细胞条件培养基(RCM)对成骨功能的调节作用.小鼠骨细胞系MLO-Y4三维回转培养72h后,收集回转条件培养基(RCM)和未回转对照组的条件培养基(CCM),用四甲基偶氮唑盐比色(MTT)法、对硝基苯磷酸(pNPP)法和流式细胞术(FCM)分别检测RCM对小鼠成骨样细胞系MC3T3-E1增殖、周期及细胞分泌碱性磷酸酶(ALP)活性的影响.采用RT-PCR方法检测RCM对MC3T3-E1成骨相关基因表达的影响.结果显示,三维随机回转72h后的MLO-Y4RCM可促进MC3T3-E1增殖:条件培养基培养MC3T3-E124h和48h后,50%RCM组比CCM组分别增加了1.62和1.60倍,差异显著(*P0.05),培养72h后,100%RCM组比CCM组增加了1.69倍,差异显著(*P0.05);细胞周期检测结果表明,条件培养基培养24、48和72h后,RCM组部分恢复CCM引起的MC3T3-E1细胞周期阻滞;MC3T3-E1的ALP活性在RCM组和CCM组之间无差异;RT-PCR检测结果表明,100%MLO-Y4条件培养基培养MC3T3-E148h后,降低了成骨相关基因ALP、Runx2、OPN、OC的表达.差异显著(*P0.05,**P0.01,***P0.001).实验结果表明,三维随机回转模拟失重培养骨细胞72h后的条件培养基促进了成骨细胞增殖,抑制了成骨相关基因表达.     

骨细胞功能研究进展

骨细胞(osteocyte)是骨组织中含量最为丰富的细胞,其寿命甚至可接近机体的寿命.骨细胞是一种活动的、具有多种功能的细胞.作为骨机械应力的直接感受器,骨细胞不仅可以将机械应力信号转化为生化信号,并且可将信号传递到骨组织其他类型细胞、调控其功能活动;此外,骨细胞还具有调整骨陷窝微环境和矿物质平衡的作用.骨细胞网络结构的完整性对于维持骨组织的正常功能至关重要.骨细胞在失重引起的骨丢失机制中也具有重要作用.     

多不饱和脂肪酸对人体的作用

多不饱和脂肪酸又叫多烯酸,是指分子结构中含有２个或２个以上不饱和双键的脂肪酸。双键愈多,不饱和程度愈高,营养价值也愈高［１］。随着科学的发展,某些多不饱和脂肪酸对人体的作用进一步被认识,特别是以廿二碳六烯酸、廿碳五烯酸和一般植物油中的亚油酸（常与亚麻...     

膳食中高n-6/n-3多不饱和脂肪酸比值对小鼠肠道菌群的影响

目的:探讨小鼠膳食中高n-6/n-3多不饱和脂肪酸比值对肠道菌群的影响。方法:随机选取健康的30只C57/B6小鼠分为对照组(基础饲料)、花生四烯酸组(基础饲料+10%花生四烯酸)、鱼油组(基础饲料+10%鱼油)。喂食16周,16周后提取小鼠肠道菌群宏基因组DNA,采用Roche 454测序技术对肠道菌群16Sr RNA基因V3-V5区域进行测序,对菌群的组成结构以及含量的变化进行分析。结果:花生四烯酸组小鼠体内厚壁菌门的含量达到(55.3±5.26)%,与对照组小鼠体内厚壁菌门的含量(30.23±8.75)%相比显著性升高(P0.05)。并且花生四烯酸组小鼠体内变形菌门的含量(3±0.762)%与对照组(1.5±0.265)%相比也显著性上升(P0.05)。结论:膳食中高n-6/n-3多不饱和脂肪酸比值会造成小鼠体内肠道菌群组成结构的失衡,导致厚壁菌门以及变形菌门的含量上升。     

破骨细胞骨吸收功能的检测方法

破骨细胞是一种多核的,具有骨吸收功能的骨组织细胞,在骨吸收过程中起着至关重要的作用.破骨细胞骨吸收功能的异常会引发一系列的临床病症,如骨质疏松症、关节置换术后假体松动、骨硬化症和牙周病变等.破骨细胞骨吸收功能的进一步研究对于各类骨疾病的防治具有重要的意义.然而破骨细胞骨吸收功能的检测方法一直以来是制约破骨细胞研究的瓶颈之一.为此,围绕破骨细胞骨吸收功能的检测方法做一综述.     

多不饱和脂肪酸的研究进展

多不饱和脂肪酸(PUFA)是人类的必需脂肪酸,对人体有重要的生理功能,能调节人体的脂质代谢、治疗和预防心脑血管疾病、抗癌、对抗肥胖,促进生长发育和提高幼体的成活率等．综述了PUFA的生理作用和来源的研究进展,特别是在对抗肥胖、促进生长发育、提高幼体的成活率等方面的研究情况进行了阐述．     

破骨细胞伪足小体的结构和功能

破骨细胞是一种由造血干细胞分化而来的具有骨吸收功能的多核细胞。破骨细胞的骨吸收功能与其肌动蛋白骨架的完整性有关。研究表明,破骨细胞肌动蛋白骨架的基本结构为伪足小体（podosome）。在破骨细胞分化的不同阶段,伪足小体呈现不同的形态结构。伪足小体的形成过程及结构完整性直接影响着破骨细胞的分化及其骨吸收活性。深入研究伪足小体的结构和功能可为探索破骨细胞的骨吸收机制和寻找骨骼疾病药物作用靶点提供新的思路。该文将围绕破骨细胞伪足小体的结构、功能及其调节机制进行综述。     

海洋细菌产生的多不饱和脂肪酸

鱼油的主要成分为二十碳五烯酸和二十二碳六烯酸,这些多不饱和脂肪酸具有多方面的生理活性。近年来在海洋细菌中发现这些多不饱和脂肪酸的存在,海洋细菌很可能是这些多不饱和脂肪酸的原始生产之一。对其生物合成的深入研究表明,海洋细菌多不饱和脂肪酸的合成不同于其他生物的不饱和脂肪酸的合成机制,合成过程中不涉及重要的脂肪酸脱氢和延长机制,其合成由一种多聚乙酰合成酶(PKS)催化。     

Order from disorder, corralling cholesterol with chaotic lipids. The role of polyunsaturated lipids in membrane raft formation

A myriad of health benefits including the prevention of cancer and heart disease accompanies consumption of polyunsaturated fatty acids (PUFA). Of special importance is the omega-3-PUFA docosahexaenoic acid (DHA), with 22 carbons and six double bonds that constitute the most highly unsaturated fatty acid naturally occurring. Our experiments target the membrane as a likely site of action and focus upon the interaction of cholesterol with PUFA-containing phospholipids. They support the idea that steric incompatibility of the rigid steroid moiety for highly disordered PUFA chains promotes lateral segregation of lipids into PUFA-rich/sterol-poor and PUFA-poor/sterol-rich regions. Solid state 2H NMR and X-ray diffraction demonstrate that the solubility of cholesterol is low in polyunsaturated bilayers. In mixed membranes of phosphatidylethanolamine (PE) with the lipid raft-forming molecules sphingomyelin (SM) and cholesterol, diminished affinity of the sterol for 1-[2H31]palmitoyl-2-docosahexaenoylphosphatidylethanolamine ([2H31]16:0-22:6PE) relative to 1-[2H31]palmitoyl-2-oleoylphosphatidylethanolamine ([2H31]16:0-18:1PE) is identified by 2H NMR order parameters. Here, lies the origin of a potential biological advantage of the relatively modest increase in PUFA content of plasma membranes that would be conferred by dietary supplementation. We hypothesize that the enhanced propensity to form SM-/cholesterol-rich rafts as well as PUFA-rich/cholesterol-poor microdomains would modify the function of proteins for which these respective regions provide a platform.     

共轭亚油酸甘油酯对大鼠血清脂肪酸组成影响的研究

目的：研究共轭亚油酸甘油酯对高脂饮食大鼠血清脂肪酸组成变化的影响;方法：60 只SD 大鼠随机分为5 组：正常对照 组、高脂对照组,共轭亚油酸甘油酯低（2 g/kg·bw）、中（4 g/kg·bw）、高（6 g/kg·bw）剂量组,除基础对照组外其余各组均喂饲高脂 饲料,建立高脂模型,以灌胃方式给予受试物,6 周后取血清测定其脂肪酸组成。采用一步法直接对血清中脂肪酸进行甲酯化,气 相色谱毛细管柱分离,氢火焰离子化检测器（FID）定性定量分析;结果：共轭亚油酸甘油酯低、中、高剂量组大鼠血清中单不饱和 脂肪酸含量为25.66%,18.74%,17.72%,与高脂对照组相比显著性下降（P<0.01）,各剂量组饱和脂肪酸和多不饱和脂肪酸含量分 别为48.08%,48.52%,51.15%和27.03%,29.28%,31.13%,与高脂对照组相比显著性升高（P<0.01）。几种代表性脂肪酸如各剂量 组中的油酸、亚油酸与高脂对照组相比分别增加了26.48%,41.56%,51.26% 和9.18%,8.61%,8.73%,各剂量组中棕榈酸与高脂对 照组相比降低了5.28%, 8.80%, 10.92%。结论：共轭亚油酸甘油酯能够增加大鼠血清中饱和脂肪酸和多不饱和脂肪酸的含量,减 少单不饱和脂肪酸含量,改变血清脂肪酸组成。     

A low omega-6 polyunsaturated fatty acid (n-6 PUFA) diet increases omega-3 (n-3) long chain PUFA status in plasma phospholipids in humans

This study aimed to determine the effect of reducing the dietary linoleic acid (LA) intake from ~5% to <2.5% energy (%E) on n-3 long chain PUFA (LCPUFA) status in humans. Thirty-six participants followed a <2.5%E LA diet for 4 weeks. Nutrient intakes were estimated from diet diaries and blood samples were collected for assessment of fatty acid composition in plasma and erythrocyte phospholipids. LA intakes were reduced from 4.6%E to 2%E during the low LA intervention (P<0.001) while n-3 LCPUFA intakes were unchanged. LA and total n-6 PUFA content of plasma and erythrocyte phospholipids were significantly reduced after the low LA diet phase (P<0.001). The n-3 LCPUFA content of plasma phospholipids was significantly increased after the low LA diet compared to baseline (6.22% vs. 5.53%, P<0.001). These data demonstrate that reducing LA intake for 4 weeks increases n-3 LCPUFA status in humans in the absence of increased n-3 LCPUFA intake.     

Low dietary fish-oil threshold for myocardial membrane n-3 PUFA enrichment independent of n-6 PUFA intake in rats

Long chain n-3 PUFA docosahexaenoic acid (DHA) is important for heart and brain function. Investigations of biologically plausible mechanisms using animal models associate cardioprotection with DHA incorporation into myocardial membranes that are largely derived from supra-physiological fish oil (FO) intake. We measured the incorporation of DHA into myocardial membranes of rats from low dietary FO intake within human dietary range and quantitatively assessed the influence of dietary n-6 PUFA. With rats fed diets containing 0.16%–5% FO, equal to 0.12%–8.7% energy (%en) as eicosapentaenoic acid (EPA) and DHA (EPA+DHA), and either 1.5%en or 7.5%en n-6 PUFA (linoleic acid) for four weeks, dietary n-6:n-3 PUFA ratios ranged from 74 to 0.3. Myocardial DHA concentration increased in a log-linear fashion with a dietary threshold of 0.019%en as EPA+DHA and half maximal dietary [EPA+DHA] equal to 0.29%en (95% CI, 0.23–0.35). Dietary linoleic acid intake did not influence myocardial DHA. Myocardial membranes are sensitive to absolute dietary intake of long chain n-3 PUFA at low %en in the rat, equivalent to a human intake of one meal of fatty fish per week or less. The dietary ratio of n-6:n-3 PUFA has no influence on long chain n-3 PUFA cellular incorporation from dietary fish oil.     

Metabolic engineering of Pseudomonas putida for production of docosahexaenoic acid based on a myxobacterial PUFA synthase

Long-chain polyunsaturated fatty acids (LC-PUFAs) can be produced de novo via polyketide synthase-like enzymes known as PUFA synthases, which are encoded by pfa biosynthetic gene clusters originally discovered from marine microorganisms. Recently similar gene clusters were detected and characterized in terrestrial myxobacteria revealing several striking differences. As the identified myxobacterial producers are difficult to handle genetically and grow very slowly we aimed to establish heterologous expression platforms for myxobacterial PUFA synthases. Here we report the heterologous expression of the pfa gene cluster from Aetherobacter fasciculatus (SBSr002) in the phylogenetically distant model host bacteria Escherichia coli and Pseudomonas putida. The latter host turned out to be the more promising PUFA producer revealing higher production rates of n-6 docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA). After several rounds of genetic engineering of expression plasmids combined with metabolic engineering of P. putida, DHA production yields were eventually increased more than threefold. Additionally, we applied synthetic biology approaches to redesign and construct artificial versions of the A. fasciculatus pfa gene cluster, which to the best of our knowledge represents the first example of a polyketide-like biosynthetic gene cluster modulated and synthesized for P. putida. Combination with the engineering efforts described above led to a further increase in LC-PUFA production yields. The established production platform based on synthetic DNA now sets the stage for flexible engineering of the complex PUFA synthase.     

Letters to the editor

The fine structure of the colonial volvocacean alga Eudorina illinoiensis (Kofoid) Pascher is described in detail, excepting the eye spot. The structure conforms closely to the Chlamydomonas type and helps confirm that certain ultrastructural features are peculiar to certain taxonomic groups, the characteristic structure of the transitional region of the flagellum being an example in this case. A spiral filament around the outer doublets of the axoneme is newly reported.

Particular attention is given to the flagellar apparatus and to the structure of the chloroplast in relation to the pyrenoid. Small stacks of thylakoids pass between the starch plates to enter the pyrenoid where they assume a tubular form. The spatial re-organisations required to achieve this are described.     

Determination of shelf life of <Emphasis Type="Italic">Spirulina platensis</Emphasis> (MI<Subscript>2</Subscript>) grown in the Philippines

Gamma linolenic acid (GLA) degradation in Spirulina followed first-order reaction kinetics. At an accelerated temperature range of 45 to 55°C, the degradation rate constants (k _r) of GLA obtained were 4.0 × 10⁻² to 8.8 × 10⁻² day⁻¹. The energy of activation (E _a) was 16.53 kcal mol⁻¹, and the Q₁₀ was 2.22. Based on 20% GLA degradation, the shelf life of sun-dried Spirulina at 30°C is 263 days or 8.6 months using the Arrhenius plot, and 258 days or 8.5 months using the Q ₁₀ approach. Presented at the 6th Meeting of the Asia Pacific Society of Applied Phycology, Manila, Philippines.     

Heterogeneous diacylglycerol acyltransferase expression enhances lipids and PUFA in Chlorella species

Algae have been explored for renewable energy, nutraceuticals, and value-added products. However, low lipid yield is a significant impediment to its commercial viability. Genetic engineering can improve the fatty acid profile of algae without compromising its growth. This study introduced the diacylglycerol acyltransferase (BnDGAT) gene from Brassica napus into Chlorella sorokiniana-I, a fast-growing and thermotolerant natural strain isolated from wastewater, which increased its intracellular lipid accumulation. Hygromycin-resistant cells were selected, and enhanced green florescence protein fluorescence was used to distinguish pure transgenic cell lines from mixed cultures. Compared to the wild type, BnDGAT expression in transgenic C. sorokiniana-I caused a threefold increase in non-polar lipid and a twofold increase in polyunsaturated fatty acids. Nile red staining reaffirmed the presence of higher intracellular lipid bodies in transgenic cells. There was a substantial alteration in the fatty acid profile of transgenic alga expressing BnDGAT. The non-essential omega 9 (C18: 1) fatty acid decreased (5%–7% from 18%), while alpha-linolenic acid, an essential omega 3 fatty acid (C18: 3), was increased (23%–24% from 11%). This study substantiates a valuable strategy for enhancing essential omega-3 fatty acids and neutral lipids to improve its nutritional value for animal feed. The increased lipid productivity should reduce the cost of producing fatty acid methyl esters (FAME). Improved FAME quality should address the clouding issues in cold regions.     

Gastrocnemius Muscle Lipids in Relation to Diet in Two Mouse Mutants, 129Re-dy and A2G-adr, with Abnormal Muscle Function

The lipids of gastrocnemius muscle from normal and dystrophic (dy) mice of the Bar Harbor, 129Re strain were studied. Animals were fed diets containing either 3.1% or 1.1% of total calories as linoleic acid. Lipid analyses were also done on muscle from a new mouse mutant, A2G-adr, which has abnormal muscle function, characterised by an arrested development of the righting response. These animals were fed the "high" linoleic acid diet only. Total lipid, triacylglycerol, and cholesterol were elevated in the 129Re-dy irrespective of the diet, whereas A2G-adr possessed significantly higher levels of cholesterol. Total phosphorus (micrograms P/g muscle) and cholesterol/phospholipid ratios were elevated in the dy strains only. Cardiolipin was raised in the dy ("low" linoleic diet) and adr muscle, whereas phosphatidylcholine was lower in the adr strain only. Linoleic acid esterified to phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine was elevated whereas arachidonic acid in phosphatidylserine was decreased in both mutants. Docosahexanoic acid (22:6) in all three dy phospholipids was decreased, independent of dietary treatment. The adr strain possessed normal levels of this fatty acid. The results specifically point to an abnormality in long-chain polyunsaturated fatty acid metabolism in gastrocnemius muscle in the 129Re-dy mutant; in the adr mutant they could reflect an abnormal increase in the number of muscle mitochondria.