Exercise: effects on hexokinase activity in red and white skeletal muscle

Single bouts of exercise increase hexokinase activity in red and white skeletal muscle of guinea pigs. Multiple bouts of exercise cause two-fold increases. In contrast to other enzymes associated with glycolysis, hexokinase activity is higher in red than in white skeletal muscle.     

17 beta-estradiol: inducer of uterine hexokinase

Administration of estradiol to ovariectomized rats induced new synthesis of uterine hexokinase which was prevented by actinomycin D, 5-fluorouracil, cycloheximide, or ethionine. The estradiol-induced increase in uterine hexokinase activity was detectable as early as 4 hours. The increase in enzyme activity was dependent upon the dose of the hormone. The evidence indicates that the increased hexokinase activity induced by 17beta-estradiol may entail an acceleration of the synthesis of certain RNA species.     

Hexokinase isoenzymes in human erythrocytes

The electrophoretic mobility of hexokinase from human erythrocytes and other tissues was studied with a new method that depends on the fluorescence of reduced nicotinamide-adenine dinucleotide phosphate for detecting enzyme activity on starch gel. The hexokinase of cord-blood erythrocytes has slightly different electrophoretic properties from that of adult red cells. Type I enzyme is split into type I(A) and type I(F); the latter is more intense in cord blood; in hemolyzates of adult blood, the activity of the two bands is usually about equal. No type II enzyme was found in cord blood. The double type I band was present in red cells from adult rabbits.     

Enzymatic basis of mannose toxicity in honey bees

Honey bees have a negligible amount of phosphomannoseisomerase, together with a high content of a hexokinase which phosphorylates mannose more efficiently than fructose or glucose. Competition at the phosphorylation level plus accumulation of mannose-6-phosphate can fully account for the toxicity of mannose in honey bees.     

啤酒废酵母制备的富铬酵母对糖尿病小鼠糖代谢的影响

[目的]研究富铬酵母对糖尿病小鼠糖代谢的影响。[方法]利用富集铬元素的啤酒废酵母制备富铬酵母,检测其基本营养指标。通过建立糖尿病小鼠模型考察富铬酵母对糖尿病小鼠的作用和有效剂量,给药试验4周后,检测血糖值、己糖激酶和苹果酸脱氢酶活性。[结果]富铬酵母的蛋白质含量为44.7%,铬含量为810.4±14.3 mg/kg,其中有机铬含量占85%以上。富铬酵母剂量在中剂量（150 mg/kg）时可显示显著的降糖能力,在高剂量时降糖能力更为显著。糖尿病小鼠在补充富铬酵母后,小鼠体内己糖激酶和苹果酸脱氢酶活力均有提高,且酶活力的增加随富铬酵母的添加量的增加而增加。[结论]在试验范围内,小鼠体内苹果酸脱氢酶活力与对照组还有较大差异,而高剂量组富铬酵母己糖激酶活力已与正常对照无显著差异。     

Oxygen-hemoglobulin dissociation curves: effect of inherited enzyme defects of the red cell

The blood of a patient with a deficiency of hexokinase in the red cells and a decreased concentration of 2, 3-diphosphoglycerate in the red cells showed an increased affinity for oxygen, whereas a patient with a deficiency of pyruvate kinase and an elevated concentration of 2, 3-diphosphoglycerate in the red cells had blood with a decreased affinity for oxygen. Defects in red cell glycolysis may alter the oxygen affinity of blood by virtue of their effect on 2, 3-diphosphoglycerate concentrations in red cells.     

绒白乳菇发酵液提取物对杨树叶枯病菌几种重要酶活性的影响

为进一步明确绒白乳菇发酵液提取物对杨树叶枯病菌生长的抑制机理,研究了该提取物对叶 枯病菌糖酵解途径中己糖激酶（HK）、丙酮酸激酶（PK）与乳酸脱氢酶（LDH）及TCA循环中琥珀酸脱氢酶（SDH）与苹果酸脱氢酶（MDH）活性的影响。结果表明:对照组菌体代谢旺盛,随处理时间的延长,5种酶活性总体呈上升趋势;提取物处理组菌体5种酶活性呈显著的下降趋势,其中处理24 h的SDH活性与处理48 h的HK、PK、MDH活性均已降至为零,而处理48 h 的LDH活性仅为对照组的14.09%。绒白乳菇发酵液提取物对5种酶的活性均具有较强的抑制作用,严重干扰了叶枯病菌菌体糖酵解途径与TCA循环的顺利进行。      

Feedback inhibition of key glycolytic enzymes in liver: action of free fatty acids

Increasing concentrations of sodium octanoate were progressively inhibitory to the activities of glucokinase, hexokinase, phosphofructokinase, and pyruvate kinase. Glucose-6-phosphate and 6-phosphogluconate dehydrogenases were also markedly inhibited. Other enzymes of carbohydrate metabolism such as lactate dehydrogenase, phosphohexose isomerase, and fructose-1,6-diphosphatase were not decreased. Among the key glycolytic enzymes, the inhibition of pyruvate kinase by the fatty acid was most marked. The biological significance of the inhibition of the key glycolytic enzymes is interpreted as a feedback inhibitory mechanism in regulation of fatty acid biosynthesis. The mechanism may function for rapid adaptation by which the organism can use the fatty acid level as a metabolic directional switch in decreasing glycolysis and turning on gluconeogenesis.     

Preparing protein microarrays by soft-landing of mass-selected ions

Intact, multiply protonated proteins of particular mass and charge were selected from ionized protein mixtures and gently landed at different positions on a surface to form a microarray. An array of cytochrome c, lysozyme, insulin, and apomyoglobin was generated, and the deposited proteins showed electrospray ionization mass spectra that matched those of the authentic compounds. Deposited lysozyme and trypsin retained their biological activity. Multiply charged ions of protein kinase A catalytic subunit and hexokinase were also soft-landed into glycerol-based liquid surfaces. These soft-landed kinases phosphorylated LRRASLG oligopeptide and D-fructose, respectively.     

Cyclic adenosine monophosphate: andromimetic action on seminal vesicular enzymes

Administration of adenosine 3',5'-monophosphate with theophylline produced testosterone-like induction of hexokinase, phosphofructokinase, pyruvate kinase, and glucose-6-phosphate dehydrogenase in the seminal vesicles of both orchidectomized and immature rats. The N(6)-O(2)'-dibutyryl analog of this cyclic nucleotide produced greater increases in vesicular enzyme activities than those induced by the parent compound. The observed enhancement of the key glycolytic enzymes and of hexose monophosphate shunt dehydrogenase was significantly inhibited by actinomycin D and cycloheximide. The evidence indicates that cyclic adenosine monophosphate may be involved as an intermediary in the action of androgenic hormones on male accessory sex organs.     

Role of the Arabidopsis glucose sensor HXK1 in nutrient,light, and hormonal signaling

Glucose modulates many vital processes in photosynthetic plants. Analyses of Arabidopsis glucose insensitive2 (gin2) mutants define the physiological functions of a specific hexokinase (HXK1) in the plant glucose-signaling network. HXK1 coordinates intrinsic signals with extrinsic light intensity. HXK1 mutants lacking catalytic activity still support various signaling functions in gene expression, cell proliferation, root and inflorescence growth, and leaf expansion and senescence, thus demonstrating the uncoupling of glucose signaling from glucose metabolism. The gin2 mutants are also insensitive to auxin and hypersensitive to cytokinin. Plants use HXK as a glucose sensor to interrelate nutrient, light, and hormone signaling networks for controlling growth and development in response to the changing environment.     

桂花己糖激酶基因家族成员的序列及表达分析

  目的  研究桂花Osmanthus fragrans己糖激酶家族成员序列特征及表达变化规律。  方法  选取3个不同花色桂花品种‘堰虹桂’O. fragrans ‘Yanhong Gui’、‘玉玲珑’O. fragrans ‘Yulinglong’、‘金球桂’O. fragrans ‘Jinqiu Gui’,从转录组数据中筛选得到HXK同源基因,进行序列分析和系统进化树分析,并利用实时荧光定量PCR检测不同品种桂花不同组织及不同发育阶段的OfHXKs基因表达情况。  结果  筛选得到OfHXK1~OfHXK4基因,分析表明不同品种桂花OfHXK1、OfHXK3和OfHXK4基因核苷酸序列相似性均高于99%。OfHXKs基因编码461~510个氨基酸残基,均包含2个保守的磷酸化作用位点和1个糖结合位点。OfHXK1和OfHXK2具有N端膜锚定结构,与拟南芥Arabidopsis thaliana AtHXK1与AtHXK2聚为一支;无跨膜区域的OfHXK3与AtHXK3亲缘关系较近,推测二者具有催化作用但无糖信号传导功能;OfHXK4蛋白腺苷结合位点处多出11个氨基酸残基,与AtHKL1和AtHKL2亲缘关系较近。4个桂花HXK基因成员在桂花1年生茎、2年生茎、嫩叶、成熟叶和花序中均有表达。随着花开放的进程,整体上OfHXK1、OfHXK3和OfHXK4基因的表达量呈现先上升后下降的趋势,而OfHXK2基因在3个品种花序发育过程中表达模式不同。  结论  根据序列分析与进化树分析,推测桂花OfHXK1~OfHXK4均具有催化己糖磷酸化的功能,OfHXK1和OfHXK2具有糖信号感知和转导的功能。OfHXK1、OfHXK3和OfHXK4基因随花序发育呈现有规律升降的表达模式,可能与其参与糖类物质代谢有关。图6表4参30     

蜂胶中4种黄酮对HepG2细胞胰岛素抵抗的改善作用

为探究蜂胶中4种黄酮成分(高良姜素、短叶松素、松属素、柯因)对胰岛素抵抗的改善作用。通过高浓度胰岛素诱导的方式建立胰岛素抵抗HepG2细胞模型;设立正常组、模型组、高良姜素处理组、短叶松素处理组、松属素处理组、柯因处理组,测定各试验组对HepG2细胞增殖、葡萄糖消耗量、糖原含量、已糖激酶(hexokinase,HK)和丙酮酸激酶(pyruvate kinase,PK)的影响。结果显示:4种黄酮成分在有效浓度范围内对胰岛素抵抗HepG2细胞增殖均无显著影响(P0.05);柯因、短叶松素作用效果不明显(P0.05);高良姜素和松属素均能不同程度地提高IR-HepG2细胞葡萄糖消耗量(glucose consumption,GC)、肝糖原含量、HK和PK活力(P0.05)。上述结果表明蜂胶中高良姜素和松属素能较好地调节IR-HepG2糖代谢,改善胰岛素抵抗,为蜂胶产品的深度开发提供参考。     

滞绿突变对大豆叶片蔗糖代谢相关基因表达的影响

为研究大豆滞绿突变对叶片衰老过程中蔗糖代谢相关基因表达产生的影响，以晋大滞绿1号及其亲本晋大74号为试验材料，测定了大豆开花后至成熟期间叶片可溶性糖含量的变化趋势，并对比了不同基因型大豆品种叶片衰老过程中蔗糖代谢相关基因的表达差异。结果表明:1)花后至成熟期间，2个供试大豆品种可溶性总糖含量均呈波动升高然后降低的变化趋势。花后14至42天，晋大滞绿1号可溶性总糖含量高于晋大74号；2)花后29至42天，晋大滞绿1号蔗糖磷酸合成酶基因SPS4，转化酶基因CInv、CWInv2以及蔗糖合酶基因SS1、SS2-2表达水平显著高于非滞绿品种晋大74号；己糖激酶和果糖激酶基因Hxk和Frk表达量也高于非滞绿品种晋大74号；3)除 SUT4-1外，其余6个蔗糖转运体基因SUTs的表达模式具有明显的品种特异性。花后29至42天，晋大滞绿1号叶片SUTs表达水平较高。综上，滞绿突变对于大豆蔗糖代谢相关基因的表达产生了明显的影响，特别是在鼓粒初期和中期等大豆籽粒形成的关键时期，滞绿品种晋大滞绿1号蔗糖代谢相关基因的表达更加活跃。     

蛋氨酸铬对鲤糖代谢相关酶活性及IR、GLUT2和SGLT基因表达的影响

为探究蛋氨酸铬(CrMet)对鲤Cyprinus carpio糖代谢相关酶活性及糖代谢相关基因表达的影响,以酪蛋白为蛋白源,豆油为脂肪源,配制7组纯化饲料,其中Cr~(3+)水平分别为0(对照)、0.1、0.2、0.4、0.8、1.6、3.2 mg/kg。选取初始体质量为(40.95±4.80)g的鲤,随机分为7组,分别投喂7种饲料,每个组设置3个重复,每个重复放60尾鱼,饲养8周后,检测其肝胰脏糖代谢相关酶活性;禁食48 h后再投喂,再投喂0、3、6、12、24、48 h时检测Cr~(3+)水平为0(对照)、0.8、3.2 mg/kg时鱼肝胰脏IR、GLUT2和肠道SGLT基因的表达量。结果表明:添加0.8 mg/kg Cr~(3+)组的组己糖激酶(HK)、丙酮酸激酶(PK)、磷酸果糖激酶(PFK)和琥珀酸脱氢酶(SDH)活力均显著高于对照组(P0.05),磷酸酵式丙酮酸激酶(PEPCK)活力显著低于对照组(P0.05);而添加Cr~(3+)并未对葡萄糖-6-磷酸脱氢酶(G6PDH)活力产生显著影响(P0.05);投喂24、48 h时,0.8 mg/kg Cr~(3+)组IR mRNA表达量显著高于0、3.2mg/kg Cr~(3+)组(P0.05);投喂3 h时,0.8、3.2 mg/kg Cr~(3+)组GLUT2 mRNA表达量均显著高于对照组(P0.05),12、24、48 h时,0.8 mg/kg Cr~(3+)组GLUT2 mRNA表达量显著高于0、3.2 mg/kg Cr~(3+)组(P0.05);而不同Cr~(3+)添加水平对鲤肠道SGLT mRNA表达量无显著性影响(P0.05)。研究表明,在饲料中添加CrMet能够提高鲤对糖的利用能力,建议Cr~(3+)添加水平为0.8 mg/kg。     

他克林对猪精子冻后质量、抗氧化能力及糖代谢的影响

旨在研究他克林对猪精液冻后质量、抗氧化能力及糖代谢的影响机制.手握法采集12头18～24月龄健康杜洛克种公猪精液,在其冷冻保存稀释液中加0.10 mmol/L他克林并冷冻保存,检测冻后精子活率、质膜完整率、顶体完整率和畸形率、线粒体膜电位、DNA完整性,同时利用试剂盒检测总抗氧能力、丙二醛含量、超氧化物歧化酶活性、总胆...     

家蚕己糖激酶基因进化分析及其多转录本在胚胎早期发育过程中的表达特性

【目的】明确家蚕（Bombyx mori）二化性品系胚胎早期发育过程中己糖激酶基因（BmHK）不同转录本的表达特性,为深入探究BmHK基因各转录本的生物学功能及揭示其在家蚕胚胎早期发育中的作用机制提供理论依据。【方法】从NCBI检索并下载家蚕等12种昆虫的HK氨基酸序列,采用MEGA 7.0的邻接法（NJ）构建基于HK氨基酸序列相似性的系统发育进化树。以家蚕二化性品系秋丰活化越年蚕卵（丙2胚胎）为材料,分别制备非滞育命运卵（ND）、滞育命运卵（DD）和即时浸酸卵（IA）,采用实时荧光定量PCR检测BmHK基因各转录本在家蚕二化性品系胚胎早期发育过程中的表达特性。【结果】BmHK基因属于HSP70-actin家族,存在4个不同的转录本,其长度分别为4610、4608、2818和1520 bp,依次命名为BmHK-a、BmHK-b、BmHK-c和BmHK-d。BmHK-a在3种蚕卵胚胎早期发育过程中整体上呈先升高后降低的变化趋势,且各时间点均表现为IA蚕卵>ND蚕卵>DD蚕卵,IA蚕卵中的BmHK-a相对表达量于发育2 h达峰值,在ND蚕卵和DD蚕卵中于发育48 h达峰值,此后其表达快速下调,至发育96 h均处于较低水平;BmHK-b在3种蚕卵中的表达水平整体上表现为IA蚕卵>ND蚕卵>DD蚕卵,3种蚕卵中的BmHK-b相对表达量变化趋势基本一致,均随发育时间推移呈逐渐上升趋势;BmHK-c在3种蚕卵中的表达差异明显,初产蚕卵中已有较高的表达水平,受精后迅速升高,至发育72 h后BmHK-c在3种蚕卵中的相对表达量再次升高;BmHK-d在3种蚕卵中的相对表达量整体上呈先升高后降低的变化趋势,且各时间点均表现为ND蚕卵>IA蚕卵>DD蚕卵,但IA蚕卵中BmHK-d表达峰值出现的时间早于DD蚕卵和ND蚕卵。【结论】BmHK-a和BmHK-d主要在家蚕胚胎早期发育（产卵或浸酸后3 d）过程中发挥重要作用;BmHK-b虽然也参与家蚕胚胎早期发育过程,但不是主要的糖代谢相关酶;BmHK-c与家蚕胚胎早期发育关系密切,通过糖酵解为胚胎发育提供能量。     

微小RNA介导东方蜜蜂微孢子虫侵染意大利蜜蜂工蜂的分子机制

【目的】通过对东方蜜蜂微孢子虫（Nosema ceranae）纯化孢子与侵染意大利蜜蜂（Apis mellifera ligustica,简称意蜂）工蜂的东方蜜蜂微孢子虫的差异表达miRNA（DEmiRNA）及其靶mRNA进行系统分析,筛选、分析和探讨病原毒力因子和侵染因子相关的DEmiRNA及调控网络,在miRNA组学层面揭示东方蜜蜂微孢子虫对意蜂的侵染机制。【方法】利用small RNA-seq（sRNA-seq）技术对东方蜜蜂微孢子虫感染7 d和10 d的意蜂工蜂中肠和东方蜜蜂微孢子虫纯化孢子（NcCK）进行深度测序,通过连续比对rRNA数据库、西方蜜蜂（Apis mellifera）基因组和东方蜜蜂微孢子虫基因组筛滤出处于侵染过程的东方蜜蜂微孢子虫（NcT1和NcT2）数据和东方蜜蜂微孢子虫孢子的测序数据。根据P≤0.05,|log₂ fold change|≥1的标准,通过比较分析筛选出各比较组中的差异表达miRNA（differentially expressed miRNA,DEmiRNA）。通过相关生物信息学软件对DEmiRNA进行表达谱分析,靶mRNA预测及功能和代谢通路注释,以及调控网络的构建与分析。通过Stem-loop RT-qPCR验证DEmiRNA的差异表达趋势及测序数据的可靠性。【结果】NcCK vs NcT1、NcCK vs NcT2和NcT1 vs NcT2比较组分别包含164、122和60个DEmiRNA。Venn分析结果显示,3个比较组共有的上调和下调miRNA分别为5和6个。上述DEmiRNA分别预测出1 885、1 733和1 524个靶mRNA。这些靶mRNA分别注释到27、25和26个功能条目,其中注释数量最多的是新陈代谢进程、催化活性、细胞进程、结合和细胞。上述靶mRNA可分别注释到84、84和84条代谢通路,其中注释数量最多的是代谢途径、核糖体和次级代谢产物生物合成。此外,对于NcCK vs NcT1、NcCK vs NcT2和NcT1 vs NcT2中的DEmiRNA,分别有35、26和12个靶向结合MAPK信号通路相关靶mRNA,分别有49、40和17个DEmiRNA靶向结合糖酵解/糖异生通路相关靶mRNA。进一步分析发现,东方蜜蜂微孢子虫的DEmiRNA参与调控蓖麻毒素B凝集素、细胞凋亡抑制因子、极管蛋白和孢壁蛋白等病原毒力因子的基因表达,以及己糖激酶、ATP/ADP移位酶、ABC转运蛋白和转录因子ste12等侵染因子的基因表达。【结论】通过对东方蜜蜂微孢子虫纯化孢子与侵染意蜂工蜂的东方蜜蜂微孢子虫进行深入细致的miRNA组学分析和探讨,解析了病原侵染过程的miRNA差异表达谱,揭示了东方蜜蜂微孢子虫可能通过调节相应miRNA的表达水平对蓖麻毒素B凝集素、细胞凋亡抑制因子、极管蛋白和孢壁蛋白等毒力因子及己糖激酶、ATP/ADP移位酶、ABC转运蛋白和转录因子ste12等侵染因子基因表达进行调控,从而适应宿主细胞内的环境并促进自身的增殖与侵染。     

中间球海胆己糖激酶基因克隆及高温-酸化胁迫对其表达影响的初步研究

为明确中间球海胆己糖激酶的序列信息和表达规律，了解高温-酸化胁迫对其表达和生物活性的影响，以中间球海胆为研究对象，利用cDNA末端快速扩增技术获得中间球海胆已糖激酶基因的全长cDNA序列（SiHK），并利用生物信息学软件分析其序列特征，分析高温-酸化胁迫下中间球海胆肠和性腺组织中SiHK基因的表达及其酶活力变化。结果表明，SiHK基因的cDNA全长2 041 bp，编码476个氨基酸，SiHK蛋白的理论等电点为6.44，蛋白质分子质量52.72 kD。生物信息学分析显示，SiHK蛋白氨基酸序列与紫球海胆HK氨基酸序列相似最高。实时荧光定量PCR结果显示，SiHK基因的表达具有组织特异性，其中，肠和性腺组织中SiHK基因的相对表达量较高，而在体腔液和管足中SiHK酶活力较高。高温-酸化胁迫处理60 d后，与对照组相比，处理组中间球海胆的肠和性腺组织中SiHK基因的相对表达量和SiHK酶活性均发生改变。由此表明，高温-酸化胁迫可能通过调控糖代谢关键酶活性对海胆的物质代谢过程产生影响。     

高盐对缢蛏生长存活、Na+/K+-ATPase活性及能量代谢相关指标的影响

以缢蛏（Sinonovacula constricta）为研究对象,设置养殖周期为60 d的对照组（20）和高盐组（30）比较实验,分别在0、10、30和60 d进行取样,检测高盐对缢蛏生长存活、鳃组织Na+/K+-ATPase（NKA）活性及肝胰腺和血清能量代谢相关指标的影响。结果表明,高盐组缢蛏在30~60 d的养殖时间段内存活率为45.1%±2.9%,壳长、壳宽、壳高和体质量特定生长率分别为0.15%/d、0.17%/d、0.13%/d和0.61%/d,均显著低于对照组。各时间点高盐组NKA活性均显著低于对照组。10和60 d时对照组缢蛏肝胰腺己糖激酶（HK）活性显著高于高盐组,血清中葡萄糖（GLU）含量、乳酸（LA）含量和乳酸脱氢酶（LDH）活性也显著高于高盐组,高盐组缢蛏无氧代谢水平低于对照组。30 d时高盐组血清中GLU含量、LA含量和LDH活性显著高于对照组,高盐组缢蛏无氧代谢水平高于对照组。0、10和30 d时高盐组肝胰腺琥珀酸脱氢酶（SDH）活性显著高于对照组,高盐胁迫下缢蛏需氧代谢水平上升。研究表明：长期高盐养殖下,缢蛏仍能保持一定的存活率和生长速度;高盐影响了缢蛏的渗透调节和能量代谢,渗透调节增加了其能量消耗。本研究为进一步培育缢蛏耐高盐新品系提供了参考资料。