稀土离子(La~(3+)、Nd~(3+)、Ho~(3+)、Yb~(3+))对人红细胞膜Na~+/K~+ ATPase作用的初步研究

本文以低渗法从新鲜健康人红细胞中制得膜Na~+/K~+ ATPase,用于研究四种鑭系稀土离子(La~(3+)、Nd~(3+)、Ho~(3+)、Yb~(3+))对该酶的影响。结果表明,四种鑭系离子都对红细胞膜Na~+/K~+ ATPase的活性有抑制作用。当反应体系中的稀土离子浓度达50μmol/L时,Na~+/K~+ ATPase的活性所剩不多;低于50μmol/L鑭系离子时,以Nd~(3+)的抑制作用最强;金属离子螯合剂能阻止稀土离子对Na~+/K~+ ATPase的抑制作用。此项研究为稀土的生物效应的理论提供了一些新的证据。     

K~+/Na~+浓度比对罗汉果悬浮细胞生长和甜苷V合成的影响

本文研究了不同K~+/Na~+浓度比对罗汉果悬浮细胞生长和甜苷V合成的影响。结果表明,合适的K~+/Na~+浓度比促进细胞生长和甜苷V生物合成。当K~+/Na~+最佳浓度比为15:1时,罗汉果悬浮细胞内K~+浓度、K~+/Na~+、S_((K,Na))值均为最大,此时细胞利用K~+效率最高,细胞生长速率最快。此外,胞外有机酸和胞内氨基酸的变化表明此浓度比例下,细胞可能通过降低TCA循环代谢强度,以及增强EMP途径和MVA途径代谢强度,从而促进萜类化合物前体乙酰Co A向罗汉果甜苷V合成。     

高盐生境中乌拉尔甘草盐离子分泌与积存格局的研究

采用植物水培方法,以乌拉尔甘草为研究材料,用不同浓度(0、80、160、320mmol·L~(-1))NaCl溶液胁迫处理乌拉尔甘草幼苗3周后,分析其叶片表面盐离子(K~+、Ca~(2+)、Na+)分泌速率的差异,并采集盐化低地草甸重盐土生境中2年生乌拉尔甘草植株,应用ICP-AES测定其不同部位(根、根状茎、茎、老叶和幼叶)中的盐离子(K~+、Na~+、Ga~(2+)、Mg~(2+))含量,探究盐离子在乌拉尔甘草叶片上的分泌格局以及盐离子在植株体内的积存格局,为完善甘草耐盐机理的研究提供依据。结果显示:(1)随着盐胁迫浓度的升高,乌拉尔甘草叶片上K~+、Ca~(2+)、Na+的分泌速率均呈增加趋势,且Na~+的分泌速率远远大于Ca~(2+)和K+的分泌速率。(2)在乌拉尔甘草各部位中,K+的积存量从大到小依次为:幼叶根根状茎茎老叶;Na~+在各个部位的积存量都十分有限,且无论地下部分还是地上部分,差异均不大;Ca~(2+)积存量由大到小依次为:老叶幼叶茎根状茎根,且老叶中Ca~(2+)的积存量显著高于其它部位。研究认为,在重盐碱地生境中,K+主要积存在幼叶中,Ga~(2+)主要积存在老叶中,植株体内各个部位Na~+的积存量很低,乌拉尔甘草表现出明显的拒Na现象;叶片分泌的主要盐离子为Na~+;乌拉尔甘草通过泌盐的方式将Na+排出体外,从而有效降低Na~+在体内的积存,这是其能够在重盐碱地生存生长的重要原因。     

升温对湖泊底泥藻细胞复苏及其生理特性的影响

为探讨藻细胞复苏过程中环境因子的作用及其细胞生理特性的变化,在连续升高温度条件下,比较了在不同N:P值的培养基中复苏藻细胞的丰度、藻群落组成动态、藻光合活性变化,同时检测了这一过程中藻细胞中Na+K+-ATPase和Ca2+Mg2+-ATPase活性的变化。结果表明:实验期间共检测到7门,62种藻,表明太湖的底泥可以作为"种源",为藻细胞的复苏提供"种子"。6℃时蓝藻就能够萌发复苏,16℃左右是最适宜藻细胞复苏的温度。在设定的温度范围内,底泥中复苏蓝藻的光合效率随着温度的升高一直增加,表明温度越高越有利于蓝藻从底泥中的萌发和复苏;但是复苏的绿藻和硅藻的光合活性一直处在被抑制状态。低N:P值培养基中复苏的藻细胞丰度远远大于其他2种培养基中复苏的藻细胞丰度,低N:P值能够显著性的激发藻细胞从底泥中的复苏。同时,低N:P比培养液中复苏藻细胞的Na+K+-ATPase和Ca2+Mg2+-ATPase活性都显著高于其他培养液中复苏藻细胞的ATPase活性;16℃时2种ATPase活性的骤然升高与最适宜藻细胞复苏的温度相吻合,而且这个温度提前于复苏藻细胞显著增加的温度(21℃)。此外,复苏藻细胞的比生长速率与Na+K+-ATPase和Ca2+Mg2+-ATPase活性都呈现显著性的线性相关(*P<0.05)。因而,藻细胞中Na+K+-ATPase和Ca2+Mg2+-ATPase活性的恢复和升高,对推动藻细胞从底泥迁移到水柱中的萌发和复苏过程具有重要的意义。     

胡杨披针形叶与宽卵形叶的渗透调节能力的差异

以胡杨披针形叶和宽卵形叶作为实验材料,对其组织水平的离子含量及不同类型细胞的离子分布特征、可溶性有机渗透调节物质的含量、液泡膜H+-ATPase的活性进行了测定。结果表明,Na+、Cl-、K+、Ca2+等离子的分布在两种叶片及不同细胞中有差异,主要表现为宽卵形叶中Na+、Cl-的含量分别为披针形叶的111.2%和118.4%,Na/K值和Na/Ca值分别为0.75和1.78,均高于披针形叶的。宽卵形叶的栅栏细胞的Na/K值和Na/Ca值较其维管束鞘细胞的高,而披针形叶的则正相反;宽卵形叶较披针形叶含有较高浓度的可溶性糖和脯氨酸,分别为514和0.751μmol·g-1FW,而甜菜碱的含量较低,为0.703μmol·g-1FW;宽卵形叶的液泡膜H+-ATPase的活性较披针形叶的高15.47%。以上结果表明胡杨宽卵形叶的渗透调节能力较强于披针形叶。本文还对两种叶片对胡杨适应其盐渍化生境的贡献进行了讨论。     

盐碱胁迫对流苏幼苗生长及离子分布的影响

采用NaCl、Na_2SO_4、NaHCO_3、Na_2CO_(3 )四种盐按不同总盐浓度(50、100、200、300 mmol·L~(-1))和比例混合为不同处理盐碱溶液,对1a生流苏幼苗进行处理,分析了幼苗生长变化、离子代谢途径。结果表明:盐碱胁迫下流苏的生长受到显著影响。流苏幼苗的相对株高、地径生长量以及生物量均随着盐碱胁迫的加重而减少;而其根冠比却不断增加。随着盐碱浓度的增加,各器官中Na~+含量显著高于对照,其排序为:叶根茎;根中K~+含量呈下降趋势,叶中K~+含量先升后降,茎K~+含量变化较平缓,其排序为:叶茎根;各器官中K~+/Na~+呈下降趋势;流苏根的K~+-Na~+选择性吸收系数S_(K, Na)值呈下降趋势,茎、叶的S_(K, Na)值呈先升后降的趋势。研究认为,流苏幼苗对低盐碱环境具有一定的主动适应性,其盐碱适应机制主要是由于根系具有补偿生长效应及叶对Na~+进行区隔化,同时也与茎、叶选择性运输K~+的能力较强有关。     

NaCl胁迫对圆柏幼苗生长和离子吸收及分配的影响

以当年生圆柏幼苗为实验材料,采用温室调控盆栽土培法研究了不同浓度NaCl(0、100、200、300mmol·L-1)胁迫21d对其生长情况及不同器官(根、茎、叶)中K~+、Na~+、Ca~(2+)和Mg~(2+)的吸收和分配的影响,以探讨圆柏幼苗对盐环境的生长适应性及耐盐机制。结果表明:(1)随着NaCl胁迫浓度的增加,圆柏幼苗生长,包括株高、地径、相对生长量以及生物量的积累均呈下降趋势,而其根冠比却增加。(2)在各浓度NaCl胁迫处理下,圆柏幼苗根、茎、叶中Na~+含量较对照均显著增加,而且叶中Na~+含量显著高于茎和根,叶中Na~+含量是根中的5倍。(3)随着NaCl胁迫浓度的升高,圆柏幼苗各器官中K~+、Ca~(2+)和Mg~(2+)含量以及K~+/Na~+、Ca~(2+)/Na~+及Mg~(2+)/Na~+比值均呈下降趋势。(4)在NaCl胁迫条件下,圆柏幼苗根系离子吸收选择性系数SK,Na、SCa,Na、SMg,Na显著提高,茎、叶离子转运选择性系数SCa,Na、SMg,Na则逐渐降低,叶中离子转运选择性系数SK,Na则随着NaCl胁迫浓度的升高显著降低,大量Na~+进入地上部,减缓了盐胁迫对根系的伤害。研究认为,圆柏幼苗的盐适应机制主要是通过根系的补偿生长效应及茎、叶对Na~+的聚积作用来实现的,同时也与根对K~+、Ca~(2+)、Mg~(2+)的选择性运输能力增强和茎、叶稳定的K~+、Ca~(2+)、Mg~(2+)的选择性运输能力有关。     

等渗NaCL和KCL胁迫对高梁幼苗生长和气体交换的影响

本文比较研究了等渗NaCl和KCl胁迫下,高粱幼苗生长及叶片离子含量、质膜相对透性和有关气体交换参数的变化。结果表明,在低浓度NaCl和KCl胁迫7天时,高粱生长、含水量和质膜相对透性与对照相比没有明显变化,而净光合速率、蒸腾速率和气孔导度已明显下降,叶肉细胞间隙CO2浓度明显增加。NaCl胁迫下叶片Na+含量成倍增加,而K+和Ca2+含量无明显变化。KCl胁迫时叶片K+含量明显增加,Ca2+含量明显下降,而Na+含量没有明显变化。随着NaCl或KCl浓度的增加,幼苗生长和叶片含水量明显下降,质膜透性和细胞间隙CO2浓度明显增加,净光合速率、蒸腾速率和气孔导度进一步下降。 NaCl胁迫下叶片Na+含量进一步增加,K+和Ca2+进一步下降,而KCl胁迫下叶片K+含量进一步 增加,Na+和Ca2+含量进一步下降。KCl对高粱生长抑制、质膜透性、Ca2+含量下降及光合气体交换参数的影响均明显大于等渗的NaCl。     

Na超负荷与Na/H交换的关系——在等容收缩离体大鼠心脏的研究

心脏富氧灌流30min稳定后随机分为四组:(1)对照组:富氧灌流75min;(2)低血流缺氧组:低血流缺氧45min后,再富氧灌流30min;(3)Ouabain组:于低血流缺氧过程中,溶Ouabain(200μmol/L)于K—H液中,余同组(2);(4)Ouabain+Amiloride组:除在低血流缺氧期给0.5mmol/L amiloride外,余同组(3)。与低血流缺氧组相比,Ouabain可引起再灌注时心肌Na的明显增加并伴有心室功能的抑制,Amiloride可明显减轻这一损害作用。这表明,Na/K ATPase活性的抑制与再灌注心肌的Na超载有关,而这一作用的机制可能是由于Na/H交换的激活所引起。     

乌本苷免疫活性物和组织中钠泵容量的关系

依赖于Na+、K+的Na+-K+-ATP酶(EC3.6.1.3,钠泵),广泛存在于哺乳类动物细胞质膜上,是催化Na+、K+跨膜主动运输的质膜酶,除维持正常的细胞内外离子浓度梯度外,对细胞能量代谢也有重要影响,许多疾病的发生是由于钠泵活性异常引起.研究证明有2种特异性的内源性钠泵抑制因子在?..     

Uptake of cesium ions by human erythrocytes and perfused rat heart: a cesium-133 NMR study

Cesium-133 NMR studies have been carried out on suspended human erythrocytes and on perfused rat hearts in media containing CsCl. The resulting spectra exhibit two sharp resonances, arising from intra- and extracellular Cs+, separated in chemical shift by 1.0-1.4 ppm. Thus, intra- and extracellular resonances are easily resolved without the addition of paramagnetic shift reagents required to resolve resonances of the other alkali metal ions. Spin-lattice relaxation times in all cases are monoexponential and significantly shorter (3-4 times) for the intracellular component. When corrections are made for the pulse repetition rate, the total intensity of the intracellular and extracellular Cs+ resonances in erythrocytes is conserved, implying total observability of the intracellular pool. The uptake of Cs+ by erythrocytes occurs at approximately one-third the reported rate for K+ and was reduced by a factor of 2 upon addition of ouabain to the sample. These results indicate that 133Cs NMR is a promising tool for studying the distribution and transport of cesium ions in biological systems and, in some cases such as uptake by cellular Na,K-ATPase, for analysis of K+ ion metabolism.     

Thallium inhibition of ouabain-sensitive sodium transport and of the (Na+ plus K+)-ATPase in human erythrocytes.

The influence of Tl+ on Na+ transport and on the ATPase activity in human erythrocytes was studied. 0.1-1.0 mM Tl+ added to a K+-free medium inhibited the ouabain-sensitive self-exchange of Na+ and activated both the ouabain-sensitive 22Na outward transport and the transport related ATPase. 5-10mM external Tl+ caused inhibition of the ouabain-sensitive 22Na efflux as well as the (Na+ plus Tl+)-ATPase. Competition between the internal Na+ and rapidly penetrating thallous ions at the inner Na+-specific binding sites of the erythrocyte membrane could account for the inhibitory effect of Tl+. An increase of the internal Na+ concentration in erythrocytes or in ghosts protected the system against the inhibitory effect of high concentration of Tl+. A protective effect of Na+ was also demonstrated on the (Na+ plus Tl+)-ATPase of fragmented erythrocyte membranes studied at various Na+ and Tl+ concentrations.     

Caclium uptake and associated adenosine triphosphatase activity in fragmented sarcoplasmic reticulum. Requirement for potassium ions.

The effects of monovalent cations on calcium uptake by fragmented sarcoplasmic reticulum have been clarified. Homogenization of muscle tissue in salt-containing solutions leads to contamination of this subcellular fraction with actomyosin and mitochondrial membranes. When, in addition, inorganic cations are contributed by the microsomal suspension and in association with nucleotide triphosphate substrates there is an apparent inhibition of the calcium transport system by potassium and other cations. However, when purified preparations were obtained after homogenization in sucrose medium followed by centrifugation on a sucrose density gradient in a zonal rotor, calcium uptake and the associated adenosine triphosphatase activity were considerably activated by potassium and other univalent cations. When plotted against the log of the free calcium concentration there was only a slight increase in calcium uptake and ATPase activity in the absence of potassium ions but sigmoid-shaped curves were obtained in 100 mM K+ with half-maximal stimulation occurring at 2 muM Ca2+ for both calcium uptake and ATPase activity. The augmentation in calcium uptake was not due to an ionic strength effect as Tris cation at pH 6.6 was shown to be inactive in this respect. Other monovalent cations were effective in the order K+ greater than Na+ greater than NH4+=Rb+=Cs+ greater than Li+ with half-maximal stimulation in 11 mM K+, 16 mM Na+, 25 mM NH4+, Rb+, and Cs+ and in 50 mM Li+. There was nos synergistic action between K+ AND Na+ ions and both calcium uptak and associated ATPase were insensitive to ouabain. Thallous ions stimulate many K+-requiring enzymes and at one-tenth the concentration were nearly as effective as K+ ions in promoting calcium uptake. The ratio of Ca2+ ions transported to P1 released remained unchanged at 2 after addition of K+ ions indicating an effect on the rate of calcium uptake rather than an increased efficiency of uptake. In support of this it was found that during the stimulation of calcium uptake by Na+ ions there was a reduction in the steady state concentration of phosphorylated intermediate formed from [gamma-32P]ATP. It is considered that there is a physiological requirement for potassium ions in the relaxation process.     

A 133Cs nuclear magnetic resonance study of endothelial Na(+)-K(+)-ATPase activity: can actin regulate its activity?

Using (133)Cs+ NMR, we developed a technique to repetitively measure, in vivo, Na(+)-K(+)-ATPase activity in endothelial cells. The measurements were made without the use of an exogenous shift reagent, because of the large chemical shift of 1.36 +/- 0.13 ppm between intra- and extracellular Cs+. Intracellularly we obtained a spin lattice relaxation time (T1) of 2.0 +/- 0.3 s, and extracellular T1 was 7.9 +/- 0.4 s. Na(+)-K+ pump activity in endothelial cells was determined at 12 +/- 3 nmol Cs+ x min(-1) x (mg Prot)[-1] under control conditions. When intracellular ATP was depleted by the addition of 5 mM 2-deoxy-D-glucose (DOG) and NaCN to about 5% of control, the pump rate decreased by 33%. After 80 min of perfusion with 5 mM DOG and NaCN, reperfusion with control medium rapidly reestablished the endothelial membrane Cs+ gradient. Using (133)Cs+ NMR as a convenient tool, we further addressed the proposed role of actin as a regulator of Na(+)-K+ pump activity in intact cells. Two models of actin rearrangement were tested. DOG caused a rearrangement of F-actin and an increase in G-actin, with a simultaneous decrease in ATP concentration. Cytochalasin D, however, caused an F-actin rearrangement different from that observed for DOG and an increase in G-actin, and cellular ATP levels remained unchanged. In both models, the Na(+)-K(+)-pump activity remained unchanged, as measured with (133)Cs NMR. Our results demonstrate that (133)Cs NMR can be used to repetitively measure Na(+)-K(+)-ATPase activity in endothelial cells. No evidence for a regulatory role of actin on Na(+)-K(+)-ATPase was found.     

Inhibition of Na+,K+-ATPase in Penaeus indicus postlarvae by lead

The plasma membrane/mitochondrial fractions of Penaeus indicus postlarvae contain Mg2+-dependent ATPase, Na+,K+-stimulated ATPase, Na+-stimulated ATPase and K+-stimulated ATPase. The Na+,K+-activated, Mg2+-dependent ATPase was investigated further in relation to different pH and temperature conditions, and at various concentrations of protein, ouabain, ATP and ions in the incubation medium. In vitro and in vivo effects of lead were studied on the enzyme activity. In vitro lead inhibited the enzyme activity in a concentration-dependent manner with an IC50 value of 204.4 microM. In correlation with in vitro studies, in vivo investigations (both concentration and time dependent) of lead also indicated a gradual inhibition in enzyme activity. A maximum decrease of 85.3% was observed at LC50 (7.2 ppm) of lead for concentration-dependent experiments. In time-dependent studies, the decrease was maximal (81.7%) at 30 days of sublethal exposure (1.44 ppm). In addition, the substrate- and ion-dependent kinetics of Na+,K+-ATPase was studied in relation to in vitro exposure of lead; these studies suggest a non-competitive type of inhibition.     

Activation of calcium transport in skeletal muscle sarcoplasmic reticulum by monovalent cations.

The rates of calcium transport and Ca2+-dependent ATP hydrolysis by rabbit skeletal muscle sarcoplasmic reticulum were stimulated by monovalent cations. The rate of decomposition of phosphoprotein intermediate of the Ca2+-dependent ATPase of sarcoplasmic reticulum was also increased by these ions to an extent that is sufficient to account for the stimulation of calcium transport and Ca2+-dependent ATPase activity. The order of effectiveness of monovalent cations tested at saturating concentrations in increasing rate of phosphoprotein decomposition is: K+, Na+ greater than Rb+, NH4+ greater than Cs+ greater than Li+, choline+, Tris+.     

Study of ADP effect on brain Na+, K+-ATPase]

A mechanism of K-insensitive, ouabain-dependent liberation of Na+ from the cell during an increase in ADP intracellular concentration is studied. It is shown that the increase in the ADP/ATP ratio does not change the Na+, K+-ATPase affinity to K+ ions and does not result in the Na-activated, K-independent ATPase reaction. ADP protects ATPase from the inhibition by ouabain which is accounted for by a decrease in the concentration of a glycoside-sensitive form of the enzyme E2-P due to a turnover of the phosphokinase step of the reaction, but not due to the binding of free Mg2+ ions. The results obtained suggest that the increase in ADP concentration within the cell activates Na-Nan exchange along Na-transporting channels of the ionic pump.     

Binding of monovalent cations to Na+,K+-dependent ATPase purified from porcine kidney. I. Simultaneous binding of three sodium and two potassium or rubidium ions to the enzyme

We previously measured the amounts of Na+ and K+ ions bound to the Na+,K+-dependent ATPase [EC 3.6.1.3] purified from porcine kidney by a modified membrane filtration method [(1979) J. Biochem. 86, 509--523]. In this study, we improved the method for measuring the amount of the active site and measured the amount of Rb+ ions (a K+ congener) bound to the ATPase as well as those of Na+ and K+ ions to get more accurate information on the K+- and Na+-binding sites. The following results were obtained. Two kinds of cation-binding sites were found to exist on the ATPase molecule. One was the Na+-binding sites (3 mol per mol of active site). Na+ ions were bound to the sites cooperatively (Hill coefficient, 2.5--3), and the apparent dissociation constant was 0.20--0.32 mM. Three moles of Na+ ions bound to the sites was displaced by 1 mol of K+ ions bound to the ATPase (phi K, 24 microM). The other was the K+-binding sites (2 mol per mol of active site). Two moles of K+, Rb+, or Na+ ions was bound to the sites cooperatively (Hill coefficient, 1.5--2), and their apparent dissociation constants were 0.044, 0.024, and 2.2 mM, respectively. We measured the amounts of Na+ and Rb+ ions bound to the ATPase in the presence of 0.8 mM NaCl and 0.13 mM RbCl, and obtained unequivocal evidence for the simultaneous binding of 3 mol of Na+ ions and 2 mol of Rb+ ions per mol of active site of the ATPase.     

Insulin regulates ionic metabolism in a fresh water teleost, anabas testudineus (bloch)

Short term effects of insulin on total brain and branchial Na+K+ ATPase, Ca2+ ATPase and Na+, K+ and Ca2+ ions were investigated in A. testudineus. The increase in brain Ca2+ ATPase after alloxan treatment may account for an increased amount of intracellular calcium required for biochemical events taking place inside the cells. Branchial Na+K+ATPase was significantly stimulated while Ca2+ ATPase significantly inhibited after alloxan treatment. This suggests that alloxan exerts its inhibitory effect on the ATP-driven Ca2+ transport via; its action on the Ca2+ pump protein rather than the membrane permeability to Ca2+. The increased activity of brain Na+K+ ATPase at 3 and 24 hr by insulin to alloxan pretreated fish may account for the stimulated co-transport of glucose and its utilization for energy requirements and the excitatory action on neurons in the brain. The elevated brain Ca2+ ATPase may be due to the role of calcium as a second messenger in hormone action. At 24 hr, the activity of branchial Na+K+ ATPase and Ca2+ ATPase in alloxan pretreated specimens was significantly stimulated by insulin. This may be due to increased synthesis of these enzyme units. Administration of insulin (lU/fish) in normal fish significantly inhibited the activity of brain and branchial Na+K+ ATPase while brain Ca2+ ATPase showed a stimulatory effect at 3 and 24 hr compared to control. Inhibition of total branchial Ca2+ ATPase activity by insulin may be due to increased Ca2+ concentration. Higher plasma glucose level in alloxan treated groups confirms the diabetic effect of alloxan. Insulin reverses this effect. The possible mechanism by which insulin controls Na+K+ ATPase activity appears to be tissue specific. The results seem to be the first report on the effect of insulin on ATPase activity in a teleost. These data are consistent with the hypothesis that insulin performs a role in hydro mineral regulation in freshwater teleosts.     

Na+, K+-ATPase activity in erythrocytes after the effect of laser radiation

An in vitro single radiation of helium-neon laser (power flux density being 2 mW/cm2 exposure--1 and 3 min) does not change the concentration of Na+ and K+, activity of Na+, K+-dependent ATPase in erythrocytes and does not affect the intensity of active Na transport through their membrane in the donor blood. The 5 min laser action decreases the level of K+ and increases that of Na+ in the erythrocytes, activates Na+, K+-ATPases and intensifies the active Na+ transport.