间歇性低氧及运动对大鼠红细胞膜Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性的影响

Wistar雄性大鼠分为常氧对照组、常氧运动组、低氧对照组、低氧运动组.分别对常氧运动、低氧运动组进行游泳训练,对低氧对照、低氧运动组进行低氧刺激.用导管法测血压和心率,用比色法测红细胞膜Na+-K+-ATP酶Ca2+-Mg2+-ATP酶活性,探讨低氧和训练对红细胞膜Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性的影响.发现低氧刺激降低大鼠红细胞膜Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶的活性,而适宜负荷运动则可提高以上两种酶的活性.结果表明,间歇性低氧运动对大鼠红细胞膜的Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性具有一定的保护作用.     

尼罗罗非鱼在淡、海水中Na~+-K~+-ATPase活性变化

用ATPase活性测试盒测定了尼罗罗非鱼鳃和肾中Na+-K+-ATPase活性在海、淡中的变化.从淡水到海水中,尼罗罗非鱼鳃和肾Na+-K+-ATPase活性分别增强了1.61倍和1.41倍;把尼罗罗非鱼由淡水直接放入海水,鳃中的Na+-K+-ATPase活性升高较剧烈,在前期(0.5 h)最高,在鱼即将死亡的后期(8 h)次之,在中期(3 h)较低;肾中Na+-K+-ATPase活性升高较缓慢,在前期较小,中期较高,后期最高.除了在淡水中,肾的Na+-K+-ATPase活性高于鳃外,在海水中以及海水的前期、中期和后期中,鳃的Na+-K+-ATPase活性都高于肾,与肾相比,鳃对肾盐协迫更为敏感.     

不同负荷游泳训练对大鼠红细胞膜的影响

目的探讨不同负荷游泳训练对大鼠的运动能力、抗氧化酶SOD的活性、脂质过氧化产物MDA含量、Na -K -ATP酶和Ca2 -ATP活性的影响。方法选取雄性SD大鼠60只,随机分为三大组,即对照组(C组,20只)、小负荷游泳训练组(L组,20只)、大负荷游泳训练组(H组,20只),测试不同负荷游泳训练对大鼠红细胞膜抗氧化能力和ATP酶活性的影响。结果小负荷的运动训练可以降低安静时红细胞膜上MDA含量,提高抗氧化酶SOD活性,大负荷的训练则产生相反的结果;力竭运动后小负荷训练组红细胞膜Na -K -ATP酶活性增加,大负荷训练组Na -K -ATP酶活性下降;力竭运动后即刻各组RBCM上Ca2 -ATP酶活性都有下降,安静时小负荷的运动训练组Ca2 -ATP酶活性提高,但没有显著性差异,大负荷的运动训练使Ca2 -ATP酶活性显著下降。结论小负荷的运动训练可以提高机体的运动能力,而大负荷的训练会对机体产生不良影响;适宜负荷的运动训练不但可以提高安静时酶的活性,而且可以提高酶活性的储备力,延缓运动性疲劳的发生及抵抗力竭运动对机体造成运动损伤。     

锌-金属硫蛋白对力竭运动大鼠肝脏和股四头肌线粒体ATPase损伤的保护作用

通过锌-金属硫蛋白(Zn-MT)的补充干预观察对大鼠肝脏和股四头肌线粒体ATPase的影响,探讨Zn-MT对线粒体ATPase保护作用的机制.用雄性SD大鼠30只,随机分为安静对照组、运动对照组、补充Zn-MT+运动组.按Bedford运动模型运动至力竭后即刻处死,取肝脏和股四头肌制备线粒体备测.测定肝脏和股四头肌线粒体Na+,K+-ATPase、Ca2+-ATPase,Mg2+-ATPase活性,并进行有关统计学检验.结果表明,Zn-MT可以抑制由于力竭运动造成的肝脏和股四头肌中Na+,K+-ATPase、Ca2+-ATPase,Mg2+-ATPase活性的下降.     

力竭性运动对大鼠骨骼肌Ca2+含量的影响

观察大鼠力竭性运动后即刻组及三小时后组(以下简称3h组)骨骼肌内Ca2+的变化情况,探讨骨骼肌Ca2+在力竭性运动后和恢复中的变化及其机制.力竭性运动后取大鼠骨骼肌,采用原子吸收分光光度计测其Ca2+浓度.运动后即刻组骨骼肌中Ca2+浓度(2068.33±752.02μg/g)比对照组(615.31±546.03μg/g)有极显著增高(P＜0.001),3 h组(734.58±258.70μg/g)与对照组无明显差异,即刻组和3 h组有极显著性差异(P＜0.001)力竭性运动即刻,骨骼肌中Ca2+浓度显著增高,恢复期内Ca2+浓度逐渐恢复.     

补充白藜芦醇对大强度运动大鼠心肌损伤的保护作用

目的:探讨RES对大强度运动心肌过氧化损伤的保护作用.方法:在活动跑台上建立大强度运动模型,测试心肌各项过氧化损伤指标.结果:大强度运动可造成大鼠心肌MDA,TNOS,iNOS,NO显著升高,TSOD,CuZn-SOD,GSH-Px,cNOS,Na+,K+-ATPase,Ca2+-ATPase显著下降.补充RES可使大鼠运动后心肌MDA,TNOS,iNOS,NO下降,TSOD,CuZn-SOD,Mn-SOD,GSH-Px,cNOS,Na+,K+-ATPase,Ca2+-ATPase升高.结论:RES可增强心肌的抗氧化能力,保持心肌酶活性.提示RES对大强度运动后心肌过氧化损伤具有保护作用.     

力竭运动对大鼠骨骼肌肌浆网钙转运的影响

采用递增负荷力竭运动方式观察急性运动对肌浆网(SR)钙转运能力的影响,发现运动后股四头肌SR Ca2+-ATPase活性和Ca2+摄取速率均明显下降,表现为SR Ca2+-ATPase活性和Ca2+摄取速率分别下降51.12%(P<0.01)和17.72%(P<0.01);而脂质过氧化物浓度则显著升高,增加48.85%(P<0.01).提示急性运动后SR Ca2+-ATP ase 活性和Ca2+摄取速率下降可能是脂质过氧化增高的结果,而Ca2+摄取功能下降可能与运动性疲劳密切相关.     

秋水仙碱对斑马鱼肝脏和鳃组织中SOD及Na~+-K~+-ATPase活性的影响

将斑马鱼(Brachyclanio rerio)暴露于秋水仙碱5个浓度组(0mg·mL-1、10.00mg·mL-1、14.12mg·mL-1、19.95mg·mL-1、28.18mg·mL-1、39.80mg·mL-1)中进行96h急性毒性试验,计算其半数致死浓度(LC50),再设置秋水仙碱3个浓度(0.67mg·mL-1、1.70mg·mL-1、4.26mg·mL-1)将斑马鱼进行21d慢性毒性实验,每7天测定肝脏和鳃中的超氧化物歧化酶(SOD)及N+-K+-三磷酸腺苷酶(Na+-K+-ATPase)活性。结果表明,秋水仙碱对斑马鱼的LC50为16.90mg·mL-1,随着秋水仙碱浓度增大和染毒时间延长,斑马鱼鳃中SOD和Na+-K+-ATPase活性均显著受到抑制,肝脏中SOD活性增加,Na+-K+-ATPase活性总体呈现抑制趋势。斑马鱼肝脏和鳃中的SOD及Na+-K+-ATPase对秋水仙碱敏感,可以作为观测指标用于评价秋水仙碱的毒性。     

蜂胶对小鼠运动能力及心肌自由基代谢的影响

目的:通过给昆明(KM)小鼠灌服蜂胶液四周并于最后一次游泳训练至力竭实验,观察训练后小鼠心肌脂质过氧化损伤程度,探讨蜂胶抗氧化、保护心肌组织、提高机体抗疲劳能力的作用.方法:KM小鼠随机分为安静对照组(A)、运动对照组(B)、运动给药组(C).记录小鼠游泳至力竭时间,并测定心肌细胞MDA含量、SOD、GSH-Px、Ca2+ -ATPase的活性.结果显示:1)C组小鼠游泳至力竭时间明显长于B组(P <0.05).2) 4周递增负荷游泳训练并于最后一次力竭后,B组小鼠运动能力下降,心肌MDA含量显著增高(P<0.05),SOD、GSH-Px、Ca2+-ATPase活性均显著下降(P<0.05).3)C组小鼠力竭运动后心肌细胞MDA含量低于B组(P<0.05),SOD 、GSH-Px 、Ca2+-ATPase活性显著高于B组(P<0.05).结论: 1)4周递增负荷训练,小鼠呈疲劳状态,心肌脂质过氧化程度明显升高.2)蜂胶可通过提高抗氧化酶活性、减少自由基的生成而降低心肌脂质过氧化损伤,具有抗氧化的作用.3)蜂胶能明显延长小鼠游泳至力竭时间,具有提高机体耐力运动能力、延缓疲劳的作用.     

烟草叶和根细胞质膜Ca2+ -ATPase特性的比较研究

测定了烟草叶和根细胞质膜Ca2+-ATPase的活性并对它们的特性进行了比较.结果表明,二者在许多方面都不尽相同.前者的最适pH为7.0,后者则为6.0,而且在一个相对较宽的pH范围内有高的活性.叶细胞质膜Ca2+-ATPase的最适反应温度为45℃,[ATP]0.5为2.2mmol/L,根细胞质膜Ca2+-ATPase则分别为40℃和1.9mmol/L.叶和根的细胞质膜Ca2+-ATPase均受Ca2+激活.曙红B(EB)对烟草叶和根细胞质膜Ca2+-ATPase有明显的抑制作用,当EB浓度达到50μmol/L后,抑制作用才不再有显著的增加.本文讨论了烟草叶和根细胞质膜Ca2+-ATPase特性出现差异的原因及其与环境的关系.     

龙柚果实总钙、可溶性Ca2+含量及Ca2+-ATPase活性的变化

研究以柚类品种龙柚为材料,对发育过程中果皮、果肉总钙和可溶性Ca~(2 )含量及Ca~(2 )-ATPase活性进行了测定.结果表明:龙柚花前至花期子房总钙含量相对较低,花后有明显上升;龙柚果皮总钙和可溶性Ca~(2 )均在果实增大期保持持续增长的趋势,此时的果肉总钙呈明显下降趋势,而对应可溶性Ca~(2 )的变化幅度较大;龙柚Ca~(2 )-ATPase活性在花前较低,花期开始上升;而果皮和果肉的可溶性Ca~(2 )含量和Ca~(2 )-ATPase活性在增大期内均有一明显增长的过程或相对较高的水平.     

地西泮对大鼠脑突触体内Ca2+水平和Ca2+, Mg2+-ATP酶活性的作用

用Quin 2法测得大鼠脑突触体内静息游离Ca2+浓度([Ca2+]i)为85±13μmol/g protein.地西泮0.1,1和10 mmol/L对静息突触体[Ca2+]i无明显影响,但能以剂量依赖方式增高65 mmol/LKCl所致突触体[Ca2+]i升高,从105±23μmol/g protein分别达到185±28,267±49和291±48μmol/gprotein.地西泮1和10 mmol/L分别使突触体Ca2+,Mg2+-ATP酶活性降低28.7%和42.5%;使Mg2+-ATP酶活性分别降低12.6%和32.8%,提示地西泮可能是通过抑制钙调素,进而抑制Ca2+,Mg2+-ATP酶活性,使突触体[Ca2+]1升高,促进神经末梢释放递质.     

Location of high affinity Ca2+-binding sites within the predicted transmembrane domain of the sarcoplasmic reticulum Ca2+-ATPase

Cation pumps bind and translocate ions with the intermediate formation of a phosphoenzyme. In spite of extensive knowledge of the primary and even secondary structures of several of these cation transport enzymes, however, no high affinity cation binding sites have yet been determined. Here we report the use of oligonucleotide-directed, site-specific mutagenesis to identify the amino acids involved in Ca2+ binding in one of these transport enzymes, the Ca2+-ATPase of sarcoplasmic reticulum. Alteration of Glu 309, Glu 771, Asn 796, Thr 799, Asp 800 or Glu 908, each of which is predicted to lie near the centre of the transmembrane domain in putative transmembrane sequences M4, M5, M6 and M8 resulted in complete loss of Ca2+ transport function and of Ca2+-dependent phosphorylation of the enzyme by ATP. Phosphorylation of each of the mutant enzymes with inorganic phosphate was observed, however, even in the presence of Ca2+, which inhibits phosphorylation in the wild-type enzyme possessing an intact high affinity Ca2+-binding site. These results suggest that at least six polar, oxygen-containing residues lying near the centre of the transmembrane domain provide ligands for one or both of the two high affinity Ca2+ binding sites in the Ca2+-ATPase.     

仙人掌多糖对荷瘤小鼠肿瘤细胞钙泵的影响

研究了食用仙人掌多糖、药用仙人掌多糖对S180小鼠肿瘤细胞膜Ca2 -ATPase的活性影响.结果表明,二者均能明显抑制荷瘤小鼠肿瘤细胞膜上Ca2 -ATPase的活性.仙人掌多糖的这一作用改变了荷瘤小鼠细胞膜的物质、能量平衡,推测其作用可能表现为促进肿瘤细胞的凋亡.     

Ca2+、CaM及CaM拮抗剂对粟酒裂殖酵母质膜H+-ATP酶活性的影响

经差速离心,酸处理,蔗糖密度梯度离心制备粟酒裂殖酵母（Schizosaccharomyces pombe）质膜H^＋-ATP酶,以研究Ca^2＋、CaM以及CaM拮抗剂对纯化的质膜H＋-ATP酶活性的影响.结果表明Ca^2＋强烈的抑制该酶的活性,而CaM对该酶的活性没有影响,但TFP与Compound R24571这两类CaM拮抗剂又都能强烈地抑制该酶的活性.推测TEP等抑制该酶的活性不是通过与CaM结合作为CaM的拮抗剂竞争性抑制该酶的活性,而是直接对该酶发生作用或者通过磷脂或其它的CaM依赖性的膜蛋白间接对该酶产生作用.     

孕酮处理对蛋鸡蛋壳钙化过程中Ca2+-ATPase基因表达的影响

为了解孕酮对蛋鸡产蛋过程中钙离子ATP酶(Calcium-ATPase,Ca2+-ATPase)基因表达的影响,本研究以产蛋高峰期蛋鸡作为实验动物,进行孕酮处理后,利用实时荧光定量PCR技术(QRT-PCR),测定在蛋壳形成过程中输卵管子宫部Ca2+-ATPase、孕酮受体(progesterone receptor,PGR)的表达量.同时对血液中钙离子和孕酮浓度变化进行测定.结果表明:孕酮处理后,血液中钙离子浓度显著降低(P＜0.05),输卵管子宫部Ca2+-ATPase表达量极显著降低(P＜0.01),PGR mRNA表达量则显著降低(P＜0.05).结果提示,在蛋壳钙化过程中,孕酮对钙离子转运和Ca2+-ATPase表达起抑制作用.     

Ca2+-dependent and Ca2+-independent phagocytosis in human neutrophils

The phagocytic function of neutrophils is a crucial element in host defence against invading microorganisms. Two main specific receptor-mediated mechanisms operate in the phagocyte plasma membrane, one recognizing the C3b/bi fragment of complement and the other the Fc domain of immunoglobulin G (ref. 1). There is evidence that phagocytosis mediated by these receptors differs in the number and nature of the intracellular signals generated. However, the mechanisms by which receptor binding is transduced into a signal that generates the formation of the phagocyte pseudopod is not known, although extensive biochemical evidence has allowed the postulate that calcium ion gradients in the peripheral cytoplasm, by interacting with calcium-sensitive contractile proteins, initiate the process of engulfment. Using the high-affinity fluorescent calcium indicator quin2 both to measure and to buffer intracellular calcium ([Ca2+]i), we show here that in human neutrophils two mechanisms of phagocytosis coexist: a [Ca2+]i-dependent and modulated phagocytosis, triggered by activation of the Fc receptor, and a [Ca2+]i-independent mechanism triggered by the activation of the C3b/bl receptors.     

Direct addition of insulin inhibits a high affinity Ca2+-ATPase in isolated adipocyte plasma membranes.

The mechanism by which insulin regulates cellular metabolism remains unknown although indirect evidence suggests that alterations in intracellular calcium are important. More specifically, it has been proposed that insulin triggers an increase in intracellular calcium which is responsible for the subsequent modification of metabolic activities. The cell maintains a large electrochemical gradient for ionised calcium between the cytoplasm (less than 10(-6) M, as determined for muscle and nerve) and the extracellular environment (less than 10(-3) M). The plasma membrane may, therefore, be important in the regulation of calcium homeostasis, as a slight alteration in the processes maintaining this gradient could result in marked changes in cytoplasmic calcium. One such process is the active extrusion of calcium from the cell by a high affinity calcium-stimulated ATPase (Ca2+-ATPase). Such a mechanism has been well established in red cells and is postulated in nerve, liver and muscle. We have identified a high affinity Ca2+-ATPase in a plasma membrane-enriched subcellular fraction isolated from rat adipocytes which may provide the enzymatic basis for a calcium extrusion pump. We demonstrate here that the Ca2+-ATPase is specifically inhibited by the direct addition of physiological concentrations of insulin to the direct addition of physiological concentrations of insulin to the isolated plasma membranes. This effect suggests that direct regulation of calcium homeostasis may represent an important event in the mechanism of action of insulin.