脑梗塞患者红细胞膜泵活性与血液流变性的相关性研究

目的 :探讨脑梗塞 (CI)病人红细胞膜Na+ K+ ATPase、Ca2 + Mg2 + ATPase与血液流变学指标的相关性。方法 :对 49例非急性期的脑梗塞患者和 2 9例健康人分别测定Na+ K+ ATPase、Ca2 + Mg2 +ATPase、Ox LDL、LDL C、Cho C以及血液流变学指标。结果 :CI组全血粘度、血浆粘度、还原粘度、HCT、EAI、TK与Na+ K+ ATPase、Ca2 + Mg2 + ATPase活力的变化呈明显负相关 ;与Ox LDL、LDL C、Cho C呈明显正相关。结论 :CI患者红细胞膜泵活性的变化能引起血液粘滞性的改变 ,其作用主要是红细胞膜Na+ K+ 泵、Ca2 + 泵的活性降低 ,导致红细胞变形能力下降。     

高频喷射通气治疗PE-SWD兔血气和Na+-K+-ATPase图像分析定量研究

为了探讨高频喷射通气（HFJV）治疗海水淹溺肺水肿（PE－SWD）的作用机理,采用全自动血气酸碱分析仪和计算机图像分析系统,对海水淹溺肺水肿组（PE－SWD－G）、高频喷射通气组（HFJV－G）和正常对照组（Con－trolgropu,CG）兔PaO2、p8CO2、血氧饱和度（SSO2）和兔肺内N －K －ATPase进行自动检测和定量分析。结果表明,PE－SWD经HFJV治疗100min后,HFJV－G中的PaO2、SaO2和肺毛细血管内皮细胞中Na －K －ATPase活性比PE－SWD－G明显升高（P＜0．01或P＜0．05）,并且HFJV－G中Na －K －ATPase的3项参数（D1、D2和G6）几乎恢复到接近CG水平。HFJV－G兔PaO2和SaO2的升高与肺内Na －K －ATPase活性的恢复密切相关。HFJV对PESWD的治疗,关键在于能较好地纠正低氧血症,因而对肺内Na －K －ATPase的恢复有明显的促进作用。     

DM2肾病患者血清leptin水平与红细胞ATP酶活性的关系

目的：探讨了2型糖尿病（DM2）肾病患者血清瘦素（leptin）水平与红细胞膜Na＋K＋-ATP酶和Ca2＋Mg2＋-ATP酶活性改变参与DM2肾病发生的可能机制。方法：应用放射免疫分析和Reilni制膜法测定了40例DM2无肾病和32例DM2肾病患者血清leptin水平和红细胞膜Na＋K＋-ATP酶和Ca＋2Mg＋2-ATP酶含量,并与35名正常健康人作比较。结果：DM2无肾病组和肾病组血清leptin水平非常显著地高于正常人组（P〈0.01）和红细胞膜Na＋K＋-ATP酶和Ca2＋Mg2＋-ATP酶水平显著地低于正常人组（P〈0.01）。DM2肾病组与无肾病组亦有显著性差异（P〈0.05）。结论：DM2的发生、发展与血清leptin水平和红细胞膜Na＋K＋-ATP酶和Ca2＋Mg2＋-ATP酶的活性有密切的关系。     

溶血磷脂酸通过蛋白激酶C-α/钙离子途径促进大鼠星形胶质细胞增殖

目的：了解溶血磷脂酸（LPA）对星形胶质细胞（AS）活化增殖的影响及其作用机制。方法:原代培养的新生SD大鼠前脑星形胶质细胞分为对照组、PKC-α激动剂（PMA）组、LPA组、PKC-α抑制剂（Ro31-8220）组、Ro31-8220＋PMA组和Ro31-8220＋LPA组，用二苯基四氮唑溴盐（MTT）比色法以及流式细胞仪（FCM）检测AS增殖。以Fura－2/AM标记细胞内钙离子，紫外分光光度计检测其浓度。用蛋白印迹法检测细胞内PKC-α表达。结果:LPA和PMA能明显促进AS的增殖，并增加细胞内PKC-α的表达以及细胞内钙离子（［Ca2+］i）浓度（P<0.01）；Ro31-8220预处理后，显著缓解LPA促AS增殖程度和［Ca2+］i浓度的增加（P<0.01），［Ca2+］i浓度的变化与PKC-α表达量的变化呈正相关。结论:LPA通过PKC-α/Ca2+途径促进AS的增殖活化。     

不同环境中应用猪眼玻璃体液成分浓度变化推断死亡时间的研究

目的 研究在不同环境中大出血死亡猪眼玻璃体液成分变化及其与死亡时间（postmorteminterval,PMI）的关系。方法 放血处死家猪后取192只猪眼随机分为A、B两组,分别在避光、温度（15±2）℃、湿度（50±5）%的空气中以及浸没于环境温度为（15±2）℃双蒸水的条件下放置2～96 h,采集玻璃体液,运用全自动生化检测仪、超高效液相色谱分析仪（Ultra Performance Liquid Chromatography,UPLC）检测K＋、Na＋、Cl–及次黄嘌呤（hypoxanthine,Hx）浓度;采用SPSS13.0统计软件对检测数据进行回归分析。结果 眼球置于空气中时,玻璃体液中K＋、Hx与PMI相关性较高,回归方程为：PMI=-11.467＋1.954[K＋]-0.017[K＋]2＋0.511[Hx]（R2=0.858）;眼球置于双蒸水时,玻璃体液中Na＋、Cl–与PMI相关性较高,回归方程为：PMI=144.439-1.636[Na＋]＋0.007[Na＋]2-0.961[Cl-]＋0.005[Cl-]2（R2=0.622）。结论 玻璃体液K＋、Hx浓度变化可作为推断死后位于空气中的尸体较为稳定的指标;玻璃体液Na＋、Cl–浓度变化可能对死后抛入淡水尸体的PMI推断具有参考价值。     

大鼠肾缺血与再灌时近曲小管上皮细胞内钙含量变化

本实验采用细胞化学方法结合EDX微区分析及生化测试方法，从细胞水平研究了大鼠肾近曲小管上皮细胞内钙含量在缺血-再灌流损伤过程中的改变情况，并探讨了其机理。结果表明：缺血1h，可致细胞内钙含量增高，质膜Ca2 -ATPase活性下降；膜通透性无明显变化。再灌流期，细胞内钙含量持续增高；质膜Ca2 －ATPase活性在再灌流早期（2～4h）基本恢复至正常，在再灌流晚期（12～24h）下降；再灌流期膜通透性逐渐增大。肾小管上皮细胞内钙含量增加在缺血期、再灌流早期及晚期某机制并不相同。     

血管生成素-1激活tyrosine kinase／PI3K增加血管内皮细胞[Mg^2＋]i

目的：探讨血管生成素-1（Ang-1）对人脐静脉内皮细胞（HUVECs）内游离镁离子浓度（[Mg^2＋]i）的调节机制。方法：我们采用荧光指示剂mag—fura-2，运用PTi阳离子测定系统动态测HUVECs的[Mg^2＋]i。结果：Ang-1诱导的[Mg^2＋]i增加与细胞外Mg^2＋浓度无关。Ang-1诱导的[Mg^2＋]i增加与细胞内Ca^2＋浓度无关。经酪氨酸激酶阻断剂（tyrphostin A23 和 genistein），磷脂酰3激酶阻断剂（wortmannin和LY294002）预处理，均显著阻断Ang-1诱导的[Mg^2＋]i增加。但经活化丝裂原激活激酶阻断剂（SB202190和PD98059）预处理，不能阻断Ang-1诱导的[Mg^2＋]i增加。结论：Ang-1通过酪氨酸激酶／磷脂酰3激酶信号传递途径使细胞内的Mg^2＋库释放Mg^2＋，从而增加HUVECs的[Mg^2＋]i。     

生长抑素对人脐静脉内皮细胞Ca2+及其膜通道的调控研究

目的：观察生长抑素(SOM)对人脐静脉内皮细胞(HUVEC)膜上Ca2 通道及脑浆内游离Ca2 浓度([Ca2 ]i)的调控作用。方法：应用共聚焦激光扫描显微术(CLSM)和膜片钳单通道记录技术对体培养HUVEC脑浆内[Ca2 ]i、膜上Ca2 通道开放情况进行观察。结果：SOM促使HUVEC脑浆内[Ca2 ]i明显升高。胞膜上Ca2 通道的开放出现l-2min的潜伏期后也开放但其开放概率明显降低。结论：SOM对HUVEC脑浆内[Ca2 ]i的调节可能首先通过细胞内储存的Ca2 释放及稍后的细胞膜上Ca2 通道开放，从而达到其调节效应。     

白细胞衍生的白三烯对实验性大鼠肾小球细胞的增殖作用

在大鼠肾毒性血清性肾炎（ＮＳＮ）的发病早期，肾小球合成的白三烯（ＬＴ）增高，伴有肾小球增殖性核抗原（ＰＣＮＡ）（＋）细胞及肾小球增殖活性（ＧＰＡ）升高。用花生四烯酸５－脂氧化酶抑制剂ＭＫ８８６处理肾炎大鼠，可抑制肾小球ＬＴＢ４的合成及ＰＣＮＡ（＋）细胞、ＧＰＡ的升高。用Ｘ线照射去除外周血白细胞，也可抑制ＰＣＮＡ（＋）细胞及ＧＰＡ的升高。由于肾小球合成的ＬＴ来源于浸润的白细胞，本研究证实肾小球白细胞     

双歧杆菌脂磷壁酸激活巨噬细胞活性机制的研究

目的：从信号分子PKC和细胞内游离Ca^2 ([Ca^2 ]i)这一途径探索青春型双歧杆菌的脂磷壁酸（lipoteichoic acid,LTA)激活小鼠腹腔巨噬细胞的机制，同时观察巨噬细胞之间缝隙连接通讯（GJIC）的变化。方法：γ-^32P ATP磷酸转移法测定巨噬细胞总PKC活性；激光共聚焦显微镜检测[Ca^2 ]i浓度的变化；激光漂白后荧光恢复技术（FRAP）观察GJIC的状态。结果：（1）LTA能明显提高巨噬细胞总PKC活性，呈剂量依赖性；（2）LTA可显著升高巨噬细胞[Ca^2 ]i的水平，并且EDTA和维拉帕米处理组、肝素和普鲁卡因处理组以及EDTA、维拉帕米、肝素和普鲁卡因处理组巨噬细胞内[Ca^2 ]i亦升高，但明显低于对照组（P＜0．01）。（3）巨噬细胞被LTA刺激后，其平均荧光恢复率明显高于对照组（P＜0．01）。结论：LTA可通过提高PKC活性，升高[Ca^2 ]i的水平以及增强GJIC的功能来激活巨噬细胞。     

Role of calcium during lipopolysaccharide stimulation of neutrophils.

This study investigated the role of intracellular calcium concentration ([Ca]i) as a possible intermediate in the lipopolysaccharide (LPS) second messenger pathway for the activation of neutrophils (polymorphonuclear leukocytes [PMNs]). Isolated PMNs were loaded with the calcium-sensitive fluorescent dye fura-2. The PMNs were stimulated with either LPS or the positive control formyl-Met-Leu-Phe (fMLP). As expected, PMN exposure to fMLP increased [Ca]i. However, LPS stimulation did not induce any detectable changes. Depletion of intracellular Ca stores with thapsigargin, or extracellular Ca with EGTA, significantly inhibited the upregulation of the CD11b/CD18 integrin in response to fMLP but not LPS. We conclude that [Ca]i is not an early intermediate in the second-messenger pathway for the activation of PMNs by LPS.     

心肌肽素对缺氧-再给氧损伤心肌

目的:研究多肽类物质心肌肽素对缺氧-再给氧损伤心肌细胞的保护作用。方法:复制培养心肌细胞缺氧-再给氧损伤模型,缺氧60min,再给氧30min,观察心肌肽素对细胞超微结构的影响。以ACAS570进行细胞内游离Ca^2＋的定性检测;以荧光染料Fura-2-AM定量测定细胞内游离Ca^2＋浓度;以荧光偏振法测定细胞膜脂质流动性。结果:心肌肽素能使心肌细胞线粒体超微结构的损伤减轻;可剂量依赖性地降低[Ca^2＋]_i和提高细胞膜脂质流动性;可明显减少Ca^2＋伪彩色三维图色彩值。结论:心肌肽素对缺氧-再给氧损伤心肌细胞有明显保护作用,可能与其降低[Ca^2＋]sub>i和提高细胞膜脂质流动性有关。     

肾阳虚证患者红细胞LPO、SOD和ATP酶活性的变化

目的 :探讨肾阳虚证患者红细胞LPO、SOD和ATP酶活性的特点及其意义。方法 :观察 19例肾阳虚证患者和 2 1例正常人红细胞LPO、SOD和红细胞膜Na+ K+ ATP酶、Mg2 + ATP酶、Ca2 + ATP酶、Ca2 + Mg2 + ATP酶活性的变化。结果 :与对照组比较 ,肾阳虚证患者红细胞LPO含量升高 (P <0 .0 1) ,红细胞SOD活性降低 (P <0 .0 1) ;红细胞膜Na+ K+ ATP酶活性显著升高 (P <0 .0 1) ,而Mg2 + ATP酶活性变化无显著性差异 ,Ca2 + ATP酶活性升高 (P <0 .0 1) ,Ca2 + Mg2 + ATP酶活性也显著升高 (P <0 .0 1)。结论 :肾阳虚证患者红细胞内脂质过氧化反应增强 ,而抗氧化能力降低 ,红细胞膜Na+ K+ ATP酶、Ca2 + ATP酶和Ca2 + Mg2 + ATP酶活性升高 ;本研究为理解肾阳虚证的病理生理基础提供了初步实验依据     

妊高征患者血浆ET水平与红细胞膜ATP酶活性的改变与发生肾病的关系

目的：探讨了妊高征肾病患者血浆内皮素（ET）水平和红细胞膜ATP酶活性的改变参与妊高征肾病的可能机制。方法：应用放射免疫分析和Reilni制膜法制定了32例妊高征肾病患者血浆ET水平和红细胞膜上Na^＋-K^＋-ATP酶和Ca^2＋-Mg^2＋-ATP酶活性的变化,并与70名妊高征无肾病组和35名正常孕妇作比较。结果：妊高征肾病组和无肾病组血浆ET水平非常显著地高于正常孕妇组（P〈0．01）,而红细胞膜上Na^＋K^＋-ATP酶和Ca^2＋-Mg^2＋-ATP酶的活性均显著地低于正常孕妇组（P〈0．01）,妊征肾病组与无肾病组有显著性差异（P〈0．05）。结论：妊高征肾病的发生与发展与血浆ET和红细胞膜上Na^＋-K^＋-ATP酶和Ca^2＋-Mg^2＋-ATP酶的活性有密切的关系。     

家兔实验性动脉粥样硬化血管重塑的形态定量分析

目的 :探讨AS血管重塑的几何形态变化规律。方法 :将 40只新西兰雄性家兔随机等分为正常对照组 (C组 )和动脉粥样硬化模型组 (AS组 ) ,分别于实验的第 4、8、12周末取各组家兔的胸主动脉进行定性观察及定量分析。结果 :随时间延长 ,AS组斑块面积逐渐增大 ,斑块检出率逐渐增加。管腔面积在早期斑块形成时并无改变 ,晚期则明显缩小。斑块面积及管腔面积均分别与内弹性膜包围面积 (IELA)、外弹性膜包围面积 (EELA)呈正相关关系 (P <0 .0 5 ) ,IELA和EELA间也呈显著正相关关系 (P <0 .0 1) ,但斑块面积与管腔面积间并无直线相关关系 (P >0 .0 5 )。结论 :家兔AS病变随时间延长逐渐加重 ,且在斑块形成时伴内弹性膜和外弹性膜的同时扩张 ,管腔狭窄与否主要与IELA和EELA相关。IELA和EELA可作为判断管腔狭窄及评价血管重塑的指标。     

急性肾衰竭家兔脾组织形态以及髓过氧化物酶和细胞膜泵活性的变化

目的:观察急性肾功能衰竭(ARF)家兔脾组织形态以及髓过氧化物酶(MPO)和细胞膜泵(ATPase)活性的变化,探讨脾在ARF免疫功能紊乱发病学中的作用。方法:42只家兔均分为对照组、HgCl2组、甘油组。其中HgCl2组以皮下注射1%HgCl2(1.3mL/kg)、甘油组以肌肉注射50%甘油(10mL/kg)分别复制ARF模型,均分为12h、24h、48h3个亚组。在不同时点,所有动物颈总动脉插管放血备检,测定血清反映肾功能的生化指标;制备脾组织切片,观察脾组织形态;并制备10%脾组织匀浆,检测MPO和ATPase活性。结果:模型组各时点的脾组织均有不同程度的淤血、脾小梁增多。HgCl2组与甘油组各时点的脾组织匀浆MPO活性均显著高于对照组,2组24h的MPO活性高于组内12h及48h;HgCl2组与甘油组在多个时点的Na+-K+-ATPase、Ca2+-ATPase、Mg2+-ATPase、Ca2+-Mg2+-ATPase活性均显著低于对照组,且随时间延长,各ATPase有逐渐降低的趋势,除甘油组的Mg2+-ATPase外,各组48h的ATPase均低于12h。结论:家兔ARF发展过程中,脾脏有组织学损伤,且存在中性粒细胞扣押、细胞膜泵活性降低,这可能是脾作为免疫器官导致ARF免疫功能紊乱的原因之一。     

老化红细胞变形能力与膜钙依赖中性蛋白酶活性的关系

研究表明,老化红细胞变形能力明显降低,且其降低与血红蛋白浓度增高及膜弹性降低有关［１］。红细胞膜钙依赖中性蛋白酶（Ｃａｌｐａｉｎ）和它的内源性抑制剂（Ｃａｌｐａｓｔａｔｉｎ）形成红细胞中一个蛋白水解系统,参与红细胞中的信号传导,调节细胞形状、体积和细胞膜通透性,与高血压、细胞老化等生理、病理现象密切相关。Ｃａｌｐａｉｎ可限制性水解红细胞膜骨架蛋白和其它膜内蛋白,导致红细胞损伤［２,３］。而老化红细胞变形能力降低与Ｃａｌｐａｉｎ的关系尚不清楚,为此我们检测了４２例健康人老化及年轻红细胞变形能力、Ｃ…     

Effects of ouabain on intracellular ion activities of sensory neurons of the leech central nervous system.

1. The intracellular K+, Na+, and Ca2+ of mechanosensory neurons in the central nervous system of the leech Hirudo medicinalis was measured using double-barreled ion-sensitive microelectrodes. 2. After inhibition of the Na(+)-K+ pump with 5 x 10(-4) M ouabain, the intracellular K+ activity (aKi) decreased, while the intracellular Na+ activity (aNai) increased. The input resistance decreased in the presence of ouabain. The intracellular Ca2+ increased more than one order of magnitude after ouabain addition. All changes in intracellular ion activities and membrane resistance were fully reversible. 3. When extracellular Na+ concentration ([Na+]o) was removed [replaced by tris(hydroxymethyl)aminomethane (Tris)], aNai decreased. In the absence of [Na+]o, aKi and aNai remained unchanged after inhibition of the Na(+)-K+ pump by reducing the extracellular K+ concentration ([K+]o) to 0.2 mM. The membrane resistance increased under these conditions. 4. The intracellular Ca2+ decreased or remained constant after removal of [Na+]o. Addition of ouabain in the absence of [Na+]o did not change intracellular Ca2+, which only increased after readdition of [Na+]o. 5. The relative K+ permeability (PK) measured as membrane potential change during a brief increase of the [K+]o from 4 to 10 mM, increased manyfold after addition of ouabain but only little if [Na+]o had been removed before adding ouabain. 6. The results suggest that the intracellular Na+ increase after inhibition of the Na(+)-K+ pump affects the intracellular Ca2+ level by stimulating a Nai(+)-Ca2+ exchange mechanism. The subsequent intracellular Ca2+ activity (aCai) rise may result in an increase of the membrane permeability to K+ ions.     

Calcium-sodium antagonism on the frog's heart: a voltage-clamp study.

1. In double sucrose-gap voltage-clamped frog atrial fibres the influence of [Ca]o and [Na]o on membrane current and contraction was investigated. 2. The slow (secondary) inward current varied with [Ca]o but was almost insensitive to changes in [Na]o. In contrast, the phasic (transient) contraction initiated by the slow inward current was affected by both [Ca]o and [Na]o. 3. With moderate changes of [Ca]o and [Na]o from normal, the strength of phasic contraction at a given depolarization followed the [Ca]o/[Na]2o ratio approximately. This was best seen at membrane potentials near zero level. 4. Under the same conditions, tonic (sustained) contractions associated with prolonged depolarizations were strictly correlated to the [Ca]o/[Na]2o ratio at any potential. No interrelation between tonic tension and steady-state current was found. 5. With extensive changes in [Ca]o and [Na]o, the sensitivity of both phasic and tonic tension to the [Ca]o/[Na]2o ratio declined, the negative effect of [Na]o becoming smaller than was expected from this ratio. 6. In Na-free choline-Ringer, a strong contracture developed followed by a spontaneous relaxation. Starting from the relaxed state, application of depolarizing clamps gave rise to phasic contractions with a very slow relaxation while tonic contractions were apparently lacking. 7. The results are interpreted in terms of an energy-dependent carrier mechanism exchanging one Ca for two Na ions across the cell membrane. The model implies a strong asymmetry in the rate constants governing the chemical reactions on both sides of the membrane. The system is thought to operate close to equilibrium at any potential, thereby determining the steady level of myoplasmic Ca. The equilibrium itself is considered to shift upon depolarization. Assuming that [Na]i is constant, the steady level of [Ca]i is expected to be proportional to the [Ca]o/[Na]2o ratio, the scale factor being a function of membrane potential. 8. The carrier model suggests the occurrence of a depolarization-induced inward transfer of Ca which might be involved in the generation of tonic contractions. 9. The apparent lack of tonic contractions in the absence of external Na ions may be explained by a suppression of carrier-mediated Ca influx normally occurring upon depolarization. 10. The antagonistic effects of [Ca]o and [Na]o on phasic contraction are understood as being due to alterations of the Ca pumping system rather than changes in slow inward current.