Leukotrienes and lipoxins: structures, biosynthesis, and biological effects

Arachidonic acid is released from membrane phospholipids upon cell stimulation (for example, by immune complexes and calcium ionophores) and converted to leukotrienes by a 5-lipoxygenase that also has leukotriene A4 synthetase activity. Leukotriene A4, an unstable epoxide, is hydrolyzed to leukotriene B4 or conjugated with glutathione to yield leukotriene C4 and its metabolites, leukotriene D4 and leukotriene E4. The leukotrienes participate in host defense reactions and pathophysiological conditions such as immediate hypersensitivity and inflammation. Recent studies also suggest a neuroendocrine role for leukotriene C4 in luteinizing hormone secretion. Lipoxins are formed by the action of 5- and 15-lipoxygenases on arachidonic acid. Lipoxin A causes contraction of guinea pig lung strips and dilation of the microvasculature. Both lipoxin A and B inhibit natural killer cell cytotoxicity. Thus, the multiple interaction of lipoxygenases generates compounds that can regulate specific cellular responses of importance in inflammation and immunity.     

Leukotriene B4 produces hyperalgesia that is dependent on polymorphonuclear leukocytes

Leukotriene B4, at the same intracutaneous doses as bradykinin, reduced the nociceptive threshold in the rat paw. The mechanism of leukotriene B4-induced hyperalgesia was distinguished from that of the hyperalgesia elicited by prostaglandin E2 and bradykinin by its dependence on polymorphonuclear leukocytes and independence of the cyclooxygenation of arachidonic acid.     

Endocannabinoid hydrolysis generates brain prostaglandins that promote neuroinflammation

Phospholipase A(2)(PLA(2)) enzymes are considered the primary source of arachidonic acid for cyclooxygenase (COX)-mediated biosynthesis of prostaglandins. Here, we show that a distinct pathway exists in brain, where monoacylglycerol lipase (MAGL) hydrolyzes the endocannabinoid 2-arachidonoylglycerol to generate a major arachidonate precursor pool for neuroinflammatory prostaglandins. MAGL-disrupted animals show neuroprotection in a parkinsonian mouse model. These animals are spared the hemorrhaging caused by COX inhibitors in the gut, where prostaglandins are instead regulated by cytosolic PLA(2). These findings identify MAGL as a distinct metabolic node that couples endocannabinoid to prostaglandin signaling networks in the nervous system and suggest that inhibition of this enzyme may be a new and potentially safer way to suppress the proinflammatory cascades that underlie neurodegenerative disorders.     

Leukotrienes: mediators of immediate hypersensitivity reactions and inflammation

Arachidonic acid plays a central role in a biological control system where such oxygenated derivatives as prostaglandins, thromboxanes, and leukotrienes are mediators. The leukotrienes are formed by transformation of arachidonic acid into an unstable epoxide intermediate, leukotriene A4, which can be converted enzymatically by hydration to leukotriene B4, and by addition of glutathione to leukotriene C4. This last compound is metabolized to leukotrienes D4 and E4 by successive elimination of a gamma-glutamyl residue and glycine. Slow-reacting substance of anaphylaxis consists of leukotrienes C4, D4, and E4. The cysteinyl-containing leukotrienes are potent bronchoconstrictors, increase vascular permeability in postcapillary venules, and stimulate mucus secretion. Leukotriene B4 causes adhesion and chemotactic movement of leukocytes and stimulates aggregation, enzyme release, and generation of superoxide in neutrophils. Leukotrienes C4, D4, and E4, which are released from the lung tissue of asthmatic subjects exposed to specific allergens, seem to play a pathophysiological role in immediate hypersensitivity reactions. These leukotrienes, as well as leukotriene B4, have pro-inflammatory effects.     

Regulation of bone mass in mice by the lipoxygenase gene Alox15

The development of osteoporosis involves the interaction of multiple environmental and genetic factors. Through combined genetic and genomic approaches, we identified the lipoxygenase gene Alox15 as a negative regulator of peak bone mineral density in mice. Crossbreeding experiments with Alox15 knockout mice confirmed that 12/15-lipoxygenase plays a role in skeletal development. Pharmacologic inhibitors of this enzyme improved bone density and strength in two rodent models of osteoporosis. These results suggest that drugs targeting the 12/15-lipoxygenase pathway merit investigation as a therapy for osteoporosis.     

尿素包合法纯化ARA的研究

为了提高微生物油脂中花生四烯酸的含量,采用尿素包合法,利用正交试验确定了尿素包合反应的最佳条件,并采用气相色谱分析花生四烯酸含量。试验得出最佳的工艺条件为:尿素:脂肪酸为4:1(W:W)、脂肪酸:乙醇为1:12(W:W)、包合温度为-15℃和包合时间为12h,微生物油脂中花生四烯酸含量为36.84%。通过方差分析和验证试验,数学模型可靠并且拟和较好。     

A cholesterol biosynthesis inhibitor blocks Staphylococcus aureus virulence

Staphylococcus aureus produces hospital- and community-acquired infections, with methicillin-resistant S. aureus posing a serious public health threat. The golden carotenoid pigment of S. aureus, staphyloxanthin, promotes resistance to reactive oxygen species and host neutrophil-based killing, and early enzymatic steps in staphyloxanthin production resemble those for cholesterol biosynthesis. We determined the crystal structures of S. aureus dehydrosqualene synthase (CrtM) at 1.58 angstrom resolution, finding structural similarity to human squalene synthase (SQS). We screened nine SQS inhibitors and determined the structures of three, bound to CrtM. One, previously tested for cholesterol-lowering activity in humans, blocked staphyloxanthin biosynthesis in vitro (median inhibitory concentration approximately 100 nM), resulting in colorless bacteria with increased susceptibility to killing by human blood and to innate immune clearance in a mouse infection model. This finding represents proof of principle for a virulence factor-based therapy against S. aureus.     

Biological activity of some oxygenated sterols

A group of oxygenated sterols has been identified as potent and specific inhibitors of sterol biosynthesis. The ability of these compounds to inhibit sterol synthesis in cultured cells and the ineffectiveness of cholesterol under the same conditions suggest that feedback regulation of sterol biosynthesis may be brought about by an oxygenated sterol rather than by cholesterol. The nature of the regulatory sterol may vary in different cells with their specific requirements for cholesterol as a structural component or as a precursor of other steroid products. The use of oxygenated sterols to block sterol synthesis in cultured cells provides new information regarding the role of sterol in cell membrane structure and function. For example, de novo sterol synthesis is required for DNA synthesis and cell division by some cultured cells. Studies with cultured cells, and with rats and mice in vivo, suggest that oxygenated sterols could be of value in the treatment of several important human diseases.     

肉桂酸对平邑甜茶幼苗根系呼吸速率及相关酶活性的影响

 【目的】探讨外源肉桂酸对平邑甜茶幼苗根系呼吸速率和相关酶活性的影响,为深入研究肉桂酸等酚酸类物质在苹果连作障碍中造成伤害的机理提供参考。【方法】将20 ml不同浓度的肉桂酸溶液浇至栽植平邑甜茶幼苗的营养钵中,处理后定期取样,在恒温（25℃）条件下用液相Oxy-Lab氧电极测定根系呼吸速率,用冻样测定相关酶活性。【结果】测定期内,5 mg•kg-1土、25 mg•kg-1土浓度处理使基础呼吸速率和三羧酸循环(TCA)、磷酸戊糖(PPP)途径的呼吸速率均表现初期上升随后（第3天）下降但末期有所恢复的趋势,而糖酵解途径(EMP)的呼吸速率则持续下降在末期小幅回升;125 mg•kg-1土浓度处理后基础呼吸速率及3条途径呼吸速率均呈下降趋势;磷酸果糖激酶(PFK)、苹果酸脱氢酶(MDH)和葡萄糖-6-磷酸脱氢酶(G-6-PDH)活性变化趋势分别与其对应途径呼吸速率变化相似。【结论】一定浓度的肉桂酸可抑制平邑甜茶幼苗根系基础呼吸速率以及EMP、TCA、PPP途径的呼吸速率,其中,高浓度下TCA途径呼吸速率下降更为显著。各途径对应相关酶活性与其呼吸速率变化相一致,随处理浓度的升高各酶活性均降低。     

外源Ca2+、Mg2+、Cu2+和吖啶黄素对刺梨果实维生素C合成的影响

【目的】研究二价金属离子和传统抑制剂对刺梨果实维生素C合成的调控作用。【方法】以‘贵农5号’刺梨为材料,采用离体喂饲和活体注射相结合的方法,研究Ca2+、Mg2+、Cu2+等二价金属离子及吖啶黄素、石蒜碱等有机抑制剂对刺梨果实半乳糖内酯脱氢酶（GalLDH）活性和维生素C（AsA）合成的影响。【结果】Ca2+、Mg2+两种离子对初纯化的GalLDH酶活性有一定的促进作用,Cu2+则显著抑制其活性;当用金属离子与AsA合成底物半乳糖内酯（GalL）共同喂饲果肉时,Ca2+、Mg2+两种离子未有效促进AsA合成,Cu2+处理下的刺梨果肉中AsA含量甚至显著降低;在刺梨果实发育过程中,注射Ca2+对活体果实AsA合成有明显促进作用,Mg2+的作用不明显,而Cu2+依然表现出对AsA合成的抑制作用。吖啶黄素能抑制初纯化GalLDH酶液约70%的活性,而石蒜碱的抑制作用较小（约30%）,但两者在离体喂饲处理中对AsA合成表现出相似的抑制效果;活体注射处理时,吖啶黄素在处理后期对AsA合成表现出较明显的抑制作用,石蒜碱则无此效应。【结论】外源增Ca2+可促进刺梨果实AsA合成和积累,而Cu2+和吖啶黄素则有抑制作用。     

基于专利情报分析的花生四烯酸产业研究

依托Thomson Innovation专利信息平台,以2000年—2012年花生四烯酸专利为研究对象,借助专利情报分析方法,对花生四烯酸相关专利文献进行分析。从专利视角,对国际上花生四烯酸产业的发展态势进行评价,为花生四烯酸产业发展战略提供了情报参考。     

半滑舌鳎体色白化相关突变位点的鉴定

本研究对体色相关通路基因的9个错义突变位点进行了测序调查,在体色黑化、体色白化、和体色正常的半滑舌鳎小样本群体中,发现其中3个突变位点与体色白化存在显著性相关,进一步通过扩大样本验证,显示B型内皮素受体第160位的C/T位点(ertB-160-C/T)与体色白化呈极显著性相关（P<0.01）,酪氨酸酶第473的G/A位点（tyr-473-G/A）和花生四烯酸5-脂氧合酶的第1178位的C/T位点（alox5-1178-C/T）与体色白化呈显著性相关（P<0.05）。此外,这三个突变位点分别位于蛋白Periplasmic_Binding_Protein_type1、Tyosinase C-terminal和Lipoxygenase结构域。qRT-PCR检测表明,ertB在白化半滑舌鳎中有眼侧不同颜色区域的皮肤表达没有显著差异（p>0.05）,而在无眼侧皮肤中的表达明显高于有眼侧皮肤（p<0.01）。在白化半滑舌鳎中tyr在有眼侧色素部分和非色素部分的表达水平均显著高于无眼侧（p<0.01）。此外,白化半滑舌鳎中无眼侧alox5的表达水平均显著高于有眼侧（p<0.05）,有眼侧白化区alox5的表达水平明显高于有眼侧白色皮肤（p<0.01）,首次鉴定出与半滑舌鳎养殖群体体色白化相关的突变位点,为今后半滑舌鳎的体色选育提供了分子基础。     

The metabolomics variations among rice,brown rice,wet germinated brown rice,and processed wet germinated brown rice

Germination and processing are always accompanied by significant changes in the metabolic compositions of rice. In this study, polished rice (rice), brown rice, wet germinated brown rice (WGBR), high temperature and pressure-treated WGBR (WGBR-HTP), and low temperature-treated WGBR (WGBR-T18) were enrolled. An untargeted metabolomics assay isolated 6 122 positive ions and 4 224 negative ions (multiple difference ≥1.2 or ≤0.8333, P<0.05, and VIP≥1) by liquid chromatography-mass spectrum. These identified ions were mainly classified into three categories, including the compounds with biological roles, lipids, and phytochemical compounds. In addition to WGBR-T18 vs. WGBR, massive differential positive and negative ions were revealed between rice of different forms. Flavonoids, fatty acids, carboxylic acids, and organoxygen compounds were the dominant differential metabolites. Based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, there 7 metabolic pathways (phenylalanine/tyrosine/tryptophan biosynthesis, histidine metabolism, betalain biosynthesis, C5-branched dibasic acid metabolism, purine metabolism, zeatin biosynthesis, and carbon metabolism) were determined between brown rice and rice. Germination changed the metabolic pathways of porphyrin and chlorophyll, pyrimidine, and purine metabolisms in brown rice. In addition, phosphonate and phosphinate metabolism, and arachidonic acid metabolism were differential metabolic pathways between WGBR-HTP and WGBR-T18. To sum up, there were obvious variations in metabolic compositions of rice, brown rice, WGBR, and WGBR-HTP. The changes of specific metabolites, such as flavonoids contributed to the anti-oxidant, anti-inflammatory, anti-cancer, and immunomodulatory effects of GBR. HTP may further improve the nutrition and storage of GBR through influencing specific metabolites, such as flavonoids and fatty acids.     

Stimulation of colonic secretion by lipoxygenase metabolites of arachidonic acid

Both 5-hydroperoxyeicosatetraenoic acid (5-HPETE) and 5-hydroxyeicosatetraenoic acid (5-HETE) increased the short-circuit current (Isc) in rabbit colonic mucosa mounted in vitro in Ussing chambers. Measurements of chlorine-36 fluxes indicated that the Isc response to 5-HPETE is due to stimulation of active chlorine secretion. 9-, 11-, and 12-HPETE's and leukotrienes C4 and B4 produced either very small increases in Isc or no increase. In contrast to results in rabbit colon, no HPETE, HETE, or leukotriene was effective in rabbit ileal mucosa. The effects of 5-HPETE in the rabbit colon were unaffected by mepacrine, but could be partially blocked by indomethacin. These results suggest that drugs which block both cyclooxygenase and lipoxygenase may be effective antidiarrheals in patients with colitis.     

Leukotrienes as mediators in tissue trauma

A significant increase in the production of cysteinyl leukotrienes was observed after mechanical or thermal trauma in the anesthesized rat. The amount of biliary N-acetyl-leukotriene E4, which represents a suitable indicator for blood plasma leukotrienes, was used as a measure of leukotriene generation. Cysteinyl leukotrienes were rapidly eliminated from blood plasma into bile where N-acetyl-leukotriene E4 was the major metabolite. Leukotrienes were at a much lower concentration in blood plasma than in bile and differed in the pattern of metabolites. The detected amounts of leukotrienes were sufficient to induce known phenomena associated with trauma, such as tissue edema and circulatory and respiratory dysfunction. Increased leukotriene generation appears to play an important role in the pathophysiology of tissue trauma.     

毛白杨4-香豆酸:辅酶A连接酶338Val缺失突变分析

通过对毛白杨中的4--香豆酸:辅酶A连接酶1（Pt4CL1）蛋白第338位缬氨酸进行缺失突变,从而获得了具有芥子酸催化活性的Pt4CL1蛋白突变体338dVal。与野生型4CL1蛋白活性相比较,该突变体获得了催化芥子酸的活性,比活力是(162±034) nkat/mg。同时,对4--香豆酸催化活性有轻微的降低（约降低12％）,对咖啡酸的催化活性有明显增加（约增加126％）,对阿魏酸的催化活性有部分的降低（约降低29％）,对肉桂酸的催化活性没有显著变化。该结论证实了第338位缬氨酸对底物芥子酸5位甲氧基具有空间位阻效应。该项研究为通过基因工程技术调控木质素的合成提供了新的方法。      

Synthesis of compounds with properties of leukotrienes C4 and D4 in gerbil brains after ischemia and reperfusion

6-Sulfidopeptide-containing leukotriene-like immunoreactivity was synthesized in gerbil forebrains after bilateral common carotid occlusion and reperfusion. At 5, 10, or 15 minutes of ischemia, concentrations increased significantly and became more marked on reperfusion. Immunoreactivity was highest in forebrain gray matter and was below the detection limit of the assay in brain regions remote from the zone of ischemia. In vitro experiments with vascular cells and organ cultures of cerebral arteries indicate that the cerebral blood vessel wall is not a major source of biosynthetic activity in the brain. These experiments demonstrate leukotriene biosynthesis by the brain. Because synthesis occurs during ischemia and reperfusion and because leukotrienes are potent vasoconstrictors and promoters of tissue edema, they may play a role in the pathophysiology of cerebral ischemia.     

花生四烯酸高产突变株5L罐发酵及菌丝老化研究

[目的]研究花生四烯酸高产突变株的发酵及菌丝老化特性。[方法]对1株经诱变获得的花生四烯酸高产突变株A1．13进行5L罐发酵试验,考察不同培养条件对发酵产物的影响。[结果]碳源一次性过量加入并改变培养温度,可使该菌株花生四烯酸产量提高至602．99mg／L;发酵菌丝体在4℃条件下老化15d后,花生四烯酸产量达885．16mg/L。大豆油、花生油、玉米油、葵花籽油和芝麻油5种植物油均可起到消泡作用,添加浓度4％葵花籽油效果最佳,与对照相比,菌丝生物量和花生四烯酸产量分别提高了124％和22％。[结论]该研究可为突变株A1．13工业化发酵产花生四烯酸提供理论依据和现实参考。     

红花油的营养价值与保健作用

油脂是人类赖以生存必不可缺的组成部分,其所含必须脂肪酸和非必须脂肪酸的比例应适当.食用油脂与疾病的关系已研究了半个多世纪,主要集中于心脑血管疾病及癌症等方面.通过科学选用油脂和合理食用膳食油,着重叙述红花油的医疗保健作用.     

Respiratory Response of Dormant Nectarine Floral Buds on Chilling Deficiency

Changes in main biochemical respiratory pathways in dormant nectarine floral buds were studied with nectarine trees （Prunus persica.var, nectariana cv. Shuguang） in order to determine the function of respiration in dormancy release. Oxygen-electrode system and respiratory inhibitors were used to measure total respiratory rates and rates of respiratory pathways. Results showed that chilling deficiency blocked the transition of respiratory mode, and made buds stay in a state of high level pentose phosphate pathway （PPP） and low level tricarboxylie acid cycle （TCA）. The decline of PPP and activation of TCA occurred synchronously with the release of dormancy. In addition, the inhibition of PPP stimulated a respiration increase related with TCA. It could be concluded that the function of PPP activation in dormancy release might be limited and PPP declination inducing TCA activation might be part of respiration mode transition mechanism during bud sprouting.