脑益嗪抗运动病时大鼠血浆PG和小脑Na^＋—K^＋—ATPase图？…

为探讨脑益嗪抗运动病作用机理，采用放射免疫方法和计算机图像分析系统，对运动病组和脑佃嗪药物预防组大鼠血浆ＴＸＢ２，６－Ｋｅｔｏ－ＰＧＦ１α和小脑毛细血管内皮＾＋－Ｋ＾＋－ＡＴＰａｓｅ进行定量测量和分析研究。结果表明ＣＰＧ大鼠血浆ＴＸＢ２和６－Ｋｅｔｏ－ＰＧＦ１α显著低于ＭＳＧ，而小脑毛细血管内皮细胞Ｎａ＾＋－Ｋ＾＋－ＡＴＰａｓｅ活性则明显高于。     

槲皮素磷酸钾对大鼠脑血栓形成模型血小板聚集及血浆TXB_2和6-keto-PGF_(12)水平的影响

以大鼠脑血栓形成为模型,观察槲皮素磷酯钾对大鼠脑血栓形成术24h后血小板聚集、血浆TxB2和6-keto-PGF12水平的影响,以及脑电图、脑重量和病理组织学改变。结果表明:复合血栓诱导剂1ml kg-1经颈内动脉注射能诱发大鼠同侧大脑半球内血栓形成,槲皮素磷酯酶钾10-20 mg kg-1能对抗大鼠脑血栓形成,降低体内血小板自发性聚集,抑制血浆TxB2的升高。提示槲皮素磷酸酯钾可能通过其抗血小板聚集及影响花生回烯酸代谢而发挥抗脑血栓形成作用。     

脑益嗪和毛冬青甲素对大鼠动脉环产生前列环素和血小板释放血栓素A_2的影响

脑益嗪体内给药能显著地抑制血小板聚集,但对动脉环前列环素(PGI_2)的产生和血小板血栓素A_2(TXA_2)的释放无作用,其抗血栓形成的作用看来不是通过对TXA_2/PGI_2系统的影响而发挥的;毛冬青甲素静脉注射,非常显著地抑制血小板聚集、抑制血小板TXA_2的释放,但不影响动脉环PGI_2的产生,生物测定法和放射免疫分析法所得结果相平行,提示毛冬青甲素可能是一种血栓素A_2合成酶抑制剂。     

6-酮-前列腺素F_(1α)酶免疫分析

本文报告了6-酮-前列腺素(PG)F_(1α)的酶免疫分析法。用混合酸酐法交联β-半乳糖苷酶和6-酮-PG-F_(1α)。通过双抗体沉淀法分离免疫复合物和游离6-酮-PGF_(1α)。以4-甲基伞形酮-β-D-半乳糖苷作底物测定酶促反应产物的荧光强度。该方法6-酮-PGF_(1α)最低检出值0.054 pmol(20pg),批内变异系数(CV)平均3.84％,批间CV平均6.72％,回收率平均105.2％。分别用酶免疫分析和放射免疫分析法测定同一家兔主动脉样品6-酮-PGF_(1α)含量,相关系数r=0.9474。用消炎痛和咪唑作为工具药验证了该方法在药理研究中应用的可行性。     

人心脏缺血预处理对非同一缺血部位心肌的保护作用

观察在双支经皮冠状动脉腔内成形术（PTCA）中缺血预处理对非同一缺血部位心肌的作用。14例患者均为两支冠状动脉（冠脉）病变。分别观察两支冠脉成形时心绞痛程度、狭窄冠脉相关导联心电图最大ST段抬高幅度和发生时间双冠状窦静脉血浆和血清中血栓素B2（TXB2）和6-酮─前列腺素F1α（6－keto－PGF1α）浓度。观察到两支冠脉狭窄程度及PTCA参数无明显差别，但第二支冠脉PTCA时心绞痛积分及发生时间和心电图最大ST段抬高幅度及发生时间均显著低于或迟于第一支冠脉PTCA时（P＜0.05）。第二文冠脉PTCA前后血浆6－keto－PGF1α水平均高于第一支（P＜0．05）。提示缺血预处理可以保护非同一缺血部位的心肌，其机制可能同前列环素的改变有关。     

白细胞介素—8对大鼠失血性休克血浆6—keto—PGF1α和TXB2含量的影响

目的和方法：本文观察重组人内皮细胞衍生的白细胞介素－８（ｒｈＥＤＩＬ－８）对大鼠晚期失血性休克血浆６－ｋｅｔｏ－ＰＧＦ１α和ＴＸＢ２含量的影响，并与平均动脉血压（ＭＡＢＰ）的变化作相关性分析。结果：晚期失血性休克血浆６－ｋｅｔｏ－ＰＧＦ１α含量明显降低（１０６７４±１２２６ｖｓ１５６８２±１１４２）ｎｇ／Ｌ，Ｐ＜００１，ＴＸＢ２含量明显升高（３１８３６±２６．５４ｖｓ１７４９１±２１５８）ｎｇ／Ｌ，Ｐ＜００１；给予ｒｈＥＤＩＬ－８（２５０μｇ／ｋｇ）后，血浆６－ｋｅｔｏ－ＰＧＦ１α含量明显升高（３６８４７±１５６８ｖｓ１０３７６±１３１８）ｎｇ／Ｌ，Ｐ＜００１，其血浆水平与ＭＡＢＰ变化呈明显正相关（ｒ＝０．７４６，Ｐ＜００１）；ｒｈＥＤＩＬ－８对血浆ＴＸＢ２含量却无明显影响。结论：ｒｈＥＤＩＬ－８抗晚期失血性休克作用与其促进血管内皮细胞产生和释放ＰＧＩ２有关     

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。     

Cytotoxic Factor in the Blood and Plasma of Animals During Leptospirosis

Whole blood and plasma from animals in the acute stage of leptospirosis contained a toxic factor which produced a cytopathic effect on fibroblastic L cell monolayers. Firm adsorption of cytotoxic factor (CTF) to L cells occurred within 1 h. The highest titer of CTF in plasma was reached at 24 h and declined after 48 h after the inoculation of leptospires. Toxic effects were also obtained with intracerebral inoculation of mice with plasma containing CTF. Mice showed signs of motor instability and muscular spasms shortly after inoculation with CTF. Death usually occurred within 1 h.     

Normal expression and inflammation-induced changes of Na and Na/K ATPase activity in spinal dorsal horn of the rat

We tested whether that peripheral inflammation induces changes in the spinal dorsal horn ATPase activity. Adult Sprague-Dawley rats were anesthetized (thiobarbital), the left hind paw (inflammation group; n = 15) was immersed in water at 60 degrees C for 60s, which induced a local inflammation. A control group (n = 12) was tested with water at room temperature. After 60 min of peripheral inflammation left (LDH) or right lumbar dorsal horn (RDH) were processed for total, Na/K, Na and remanent ATPase activities (nM P(i) (mgprotein)(-1) min(-1)). In control animals isoenzymatic activities were: Na (31.2%); Na/K (20.6%) and remanent (48.2%) from total ATPase activity. No LDH-RDH asymmetry was found. The inflammation group presented an ipsilateral increase of total ATPase activity in LDH (X+/-S.E.M.; 4798.9+/-601) over the RDH (3982.2+/-451; Delta+817; P<0.05). This is due to an increase in Na ATPase activity (1609.3+/-297) over RDH (1164.2+/-166; Delta+445; P<0.05). ATPase activities were increased in LDH from inflamed over the control group as follows: total (4798.9+/-601; Delta+840; P<0.05), Na/K (1298.1+/-301; Delta+483; P<0.05) and Na (1609.3+/-297; Delta+373; P<0.05). These increased ATPase activities, induced in a short time, can be considered a functional marker of nociceptive neuronal activity.     

Stimulation of Xenopus oocyte Na(+),K(+)ATPase by the serum and glucocorticoid-dependent kinase sgk1

The serum and glucocorticoid-dependent kinase-1 (sgk1) is expressed in a wide variety of tissues including renal epithelial cells. As it is up-regulated by aldosterone, it is considered to participate in the regulation of renal Na(+) reabsorption. Indeed, co-expression of sgk1 with the renal epithelial Na(+) channel (ENaC) augments Na(+) channel activity. The aim of the present study was to examine possible effects of sgk1 on Na(+)/K(+)-ATPase activity. To this end dual-electrode voltage-clamp experiments were performed in Xenopus oocytes expressing the active kinase (S422D)sgk1 or the inactive mutant (K127N)sgk1. Na(+)/K(+)-ATPase activity was estimated from the hyperpolarization (delta V(m)) and the outwardly-directed current ( I(P)) created by addition of extracellular K(+) in the presence of K(+) channel blocker Ba(2+). Both delta V(m) and I(P) were significantly larger in oocytes expressing (S422D)sgk1 than in those expressing (K127N)sgk1 or having been injected with water. I(P) was fully inhibited by ouabain. Ion-selective microelectrodes showed that the stimulation of pump current was not the result of altered cytosolic Na(+) activity or pH. The present results thus point to an additional action of sgk1 that may participate in the regulation of renal tubular Na(+) transport. Moreover, sgk1 may be involved in the regulation of Na(+)/K(+)-ATPase in extrarenal tissues.     

Na−K ATPase activity and intracellular ion concentrations in the lactating guinea pig mammary gland

Summary The intracellular sodium, potassium and chloride concentrations in slices of lactating guinea pig mammary gland have been determined by chemical analysis and the use of appropriate values for extracellular space. These ion concentrations after 1 hr incubation at 37° C in a Krebs-Ringer bicarbonate solution are 45 mM Na⁺, 138 mM K⁺ and 44 mM Cl^–, which values are in agreement with those found in fresh mammary gland slices. Inhibition of the Na–K activated ATPase cation pump system of the tissue by 10^–4 M ouabain, anoxia or cooling to 0°C causes a gain of Na⁺ and an equimolar loss of K⁺ without a significant change in chloride concentration. The effect of cooling (0°C) is reversible by reincubation at 37°C. Water content of the tissue (76.5% of wet weight) and extracellular space (40.5%) do not change under these conditions. The results permit the conclusion that the Na–K activated ATPase system is responsible for the maintenance of the intracellular Na⁺ and K⁺ concentrations, but do not support the presence of a chloride pump.     

Vascular tissue (Na,K)ATPase activity and aging in the F344 rat

Vascular tissue (heart, thoracic aorta and tail artery) was removed from Fischer 344 rats, 12, 18 and 27 months of age. The intact tissue was then used to determine total, ouabain-sensitive and ouabain-insensitive 86Rubidium (86Rb) uptakes, to provide a reflection of (Na,K)ATPase activity. These studies indicate no change in total, ouabain-sensitive nor ouabain-insensitive 86Rb uptakes into cardiac tissue isolated from these rats. However, tail artery total and ouabain-sensitive 86Rb uptake decreased with age (a 61% decrease in ouabain-sensitive 86Rb uptake in 12 vs. 27-month-old rats) without significant changes in the ouabain-insensitive 86Rb uptake. This pattern was repeated in aortic tissue with a 56% decrease in ouabain-sensitive 86Rb uptake in 12 vs. 27-month-old rats. The results of these studies support an age-related decline in (Na,K)ATPase activity in aortic and tail artery tissue without a significant change in cardiac (Na,K)ATPase activity between 12, 18 and 27-month-old Fischer 344 rats.     

Enzyme secretion by the isolated rabbit pancreas: Absence of a relation with the Na−K activated ATPase

Summary The isolated rabbit pancreas secretes protein and -amylase at a relatively constant rate during an eight-hour period. Ouabain (10^–5 and 10^–6 M) did not alter enzyme secretion, but inhibited flow (65% and 28% respectively). Acetazolamide (10^–3 M) had no effect on pancreatic enzyme secretion, while flow was inhibited by 25%. Na azide (10^–2 M) failed to affect protein and -amylase secretion. Flow was inhibited by approximately 88%. NaF (10^–2M) increased both protein and -amylase secretion, while flow remained virtually unchanged. Gassing of the bathing fluid with 95% N₂–5% CO₂ reduced protein and -amylase secretion by 57 and 64% respectively, while flow was decreased by 77%. Lowering of the sodium concentration in the bathing fluid by 85% decreased -amylase secretion by 46%. Flow was inhibited by 77%. Return to the standard solution caused initially an increase of -amylase secretion (86%), followed by a decrease. These results strongly suggest that the enzyme secretion of the pancreas is not coupled to the sodium pump, responsible for fluid and electrolyte secretion.     

Morphometric Description of the Wandering Behavior in Drosophila Larvae: Aberrant Locomotion in Na+ and K+ Channel Mutants Revealed by Computer-Assisted Motion Analysis

Wandering is a simple behavior in Drosophila larvae prior to metamorphosis. Using the Dynamic Image Analysis System (DIAS) initially developed for analyzing amoeboic movements of single cells, we have analyzed videotaped behaviors of Drosophila larvae at the wandering stage. Previous studies show that mutations in the Na⁺ channel gene paralytic (para) cause paralysis at 29°C, and mutations in the K⁺ channel β subunit gene Hyperkinetic (Hk) lead to leg-shaking under ether anesthesia. The application of DIAS revealed quantifiable abnormalities in the larval locomotion of both ion channel mutants even under “permissive” conditions. Analysis of centroid movement indicates that, compared to wild type, both Hk and para larvae crawled at a slower average speed, but at a similar peak instantaneous speed during a contraction cycle. Nevertheless, contraction in the body length was greater in mutants, implying a lower efficiency in conversion of muscular contraction to distance translocation. In addition, each mutant produced a characteristic crawling pattern distinct from the wild-type control. The larval crawling pattern was determined by periods of linear locomotion interposed by non-locomotive, “searching and decision-making” episodes, after which the crawling was resumed in a new direction. Our results demonstrate that mutations in single ion channel subunits resulted in stereotypic modifications in locomotion control and crawling patterns, and that DIAS is a powerful tool in revealing subtle differences in animal behavior and quantifying mutational effects on the interplay of discrete behavioral components.     

Occurrence and properties of a (Na+−K+)-activated ATPase in the mucosa of the rat intestine

Summary The existence of an ouabain-sensitive (Na⁺–K⁺)-activated ATPase system has been demonstrated in the total intestine of the rat. The (Na⁺–K⁺)-ATPase activity was about 10–15% of the total ATPase in 4 equal parts of the small intestine; in the colon about 35% of the total ATPase was (Na⁺–K⁺)-activated ATPase. The highest (Na⁺–K⁺)-ATPase activity has been observed in the first and second part of the small intestine, while in the colon the activity was 2–4 times higher than in the ileum.The (Na⁺–K⁺)-ATPase of rat colon required both Na⁺ (K _m=8.3 mmoles/l) and K⁺ (K _m=0.6 mmoles/l). Maximal activation of the (Na⁺–K⁺)-ATPase system required 2 mM Mg²⁺ at an ATP concentration of 2 mM. The pH optimum for (Na⁺–K⁺)-ATPase of rat colon was 7.5, while the Mg²⁺-activated ATPase activity had a pH optimum of 8.6. The (Na⁺–K⁺)-ATPase was inhibited by ouabain (pI ₅₀=3.6).The relation between the differences in (Na⁺–K⁺)-ATPase activity and Na⁺-absorption on different parts of the intestine is discussed.