对不同月龄雄性大鼠成骨细胞内钙和钙通道电流的实验研究

目的研究不同月龄的雄性大鼠成骨细胞内钙浓度及钙通道电流是否存在差异.方法采用二次酶消化法分离不同月龄(分别为1、2、3、5、7、9、11、13、15)雄性大鼠的原代成骨细胞,通过激光扫描共聚焦技术测定细胞内游离钙浓度([Ca2+]i)(以平均荧光强度表示),同时应用全细胞膜片钳技术记录成骨细胞膜钙电流(ICa)的变化.结果共聚焦结果显示随月龄增加,成骨细胞内[Ca2+]i逐渐降低,但相邻2月龄组之间细胞内[Ca2+]i无显著性差异(P＞0.05);1、2、3月龄组与11、13、15月龄组成骨细胞内[Ca2+]i有显著性差异(P＜0.05),5、7、9月龄组与各组相比均无差异(P＞0.05).应用全细胞膜片钳技术发现刺激电压为+10 mV时,2、7、13月龄组鼠ICa分别为(-392.77±97.07)pA、(-330.33±33.86)pA和(-287.68±71.01)pA,13月龄组鼠与2月龄组鼠相比,ICa明显降低(P＜0.05),而7月龄组鼠与2月龄组鼠和13月龄组鼠相比,ICa均没有明显差异(P＞0.05).结论不同月龄大鼠成骨细胞内[Ca2+]i存在差异,其机制可能与细胞膜上钙通道活性改变相关.     

玉郎伞提取物对大鼠自发性高血压的影响

目的：研究玉郎伞提取物(YLS)对自发性高血压大鼠(SHR)及正常大鼠血压(BP)的影响。方法：应用颈动脉插管测压法，连接MS2000多媒体生物信号记录系统测定静注YLS前及药后20s、30s、lmin、2min、5min时的收缩压(SP)、舒张压(DP)、平均动脉压(MP)。结果：YLS使SHR及正常大鼠血压明显降低(P＜0．01)，降压作用维持时间较短，给药后5min血压基本恢复正常；YLS对去甲肾上腺素(NA)和异丙肾上腺素（ISO)的作用无明显影响。结论：YLS对正常大鼠及SHR血压有明显的降低作用。     

hCG/ATP-induced Leydig cell calcium channel reaction at different extracellular calcium ion concentration

Objective: To study the calcium channel reaction of human Leydig cells induced by hCG/ATP at different extracellular calcium ion concentrations. Methods: The Leydig cell calcium ion concentration was examined with laser confocal microscope, when the cells were stimulated with hCG/ATP at different extracellular calcium contrations. Results: With calcium-containing extracellular fluid, the Leydig cells were sensitive to hCG stimulation and when the extracellular fluid was calcium-free, the Leydig cells did not respond to the stimulation. However, the Leydig cells did respond to ATP stimulation no matter the extracellular fluid contained calcium or not. Conclusion: In human Leydig cells, there are calcium channels sensitive to hCG and ATP. The extracellular calcium ion concentration plays an important role in the regulation of Leydig cell metabolism by hCG/ATP.     

CLINICAL NEUROPHARMACOLOGY OF CALCIUM ANTAGONISTS

Summary— Experimental and clinical data clearly demonstrate that calcium antagonists (CA) may have an action on the central nervous system (CNS). The cerebrovascular action of CA justifies their use in cerebral ischaemia, vasospasm and hypoxia. Several clinical trials have demonstrated such beneficial effects. On the other hand a number of reports indicate that CA may have a direct neuronal effect, although most of such trials have not been verified or are mere case reports. In addition, the large number of conditions susceptible to being corrected by CA is impressive: epilepsy, pain, dystonia, dyskinesia, psychiatric conditions, etc. Other papers are disconcerting that report extrapyramidal disorders induced by flunarizine and cinnarizine in the elderly, whereas nicardipine does not produce such side effects and may even alleviate some parkinsonian symptoms. In various experimental models (e.g. stroke, oedema), pharmacological effects have been shown to vary from one compound to the other. Two main questions are yet to be answered: 1) has the direct neuronal effect of CA been clearly established? 2) are the multiple clinical effects on the CNS really linked to calcium antagonism?     

Cobalt-dependent potentiation of net inward current density in Helix aspersa neurons.

A low concentration of transition metal ions Co2+ and Ni2+ increases the inward current density in neurons from the land snail Helix aspersa. The currents were measured using a single electrode voltage-clamp/internal perfusion method under conditions in which the external Na+ was replaced by Tris+, the predominant external current carrying cation was Ca2+, and the internal perfusate contained 120 mM Cs+/0 K+; 30 mM tetraethylammonium (TEA) was added externally to block K+ current. In the presence of Co2+ (3 mM) or Ni2+ (0.5 mM) inward Ca2+ currents were stimulated normally by voltage-dependent activation of Ca2+ channels. There was a 5-10% decrease in the rate of rise of the inward current. The principal effect of Co2+ and Ni2+ in increasing the current density seems to be a decrease in the rate at which the inward currents decline during a depolarizing voltage pulse. The results may be due to a decrease in a voltage-dependent or Ca(2+)-dependent outward current and/or an inhibition of Ca2+ channel inactivation. Outward current under these conditions (zero internal K+) was significant and most likely due to Cs+ efflux through the voltage-activated or Ca(2+)-activated nonspecific cation channels. Co2+ is an extremely effective blocker of this outward current. These results are not an artifact of internal perfusion or the special ionic conditions. Intracellular recording of unperfused neurons in normal Helix Ringer's solution showed that the Ca(2+)-dependent action potential duration was increased significantly by low concentrations of Co2+. This result is consistant with the Co(2+)-dependent increase in inward (depolarizing) current seen in voltage-clamp experiments.     

Influence of a calcium dependent protease inhibitor on platelet activation and secretion

Continuous proteolysis resulting in consumption of major cytoskeletal proteins may be essential for platelet activation and aggregation. In this study we evaluated the effect of a known protease inhibitor, Leupeptin, on agonist induced platelet aggregation and secretion. Platelets exposed to 10 ugs/ml of Leupeptin did not aggregate in response to the action of thrombin (0.2u/ml). However, a concentration of Leupeptin as high as 250 ugs/ml did not prevent arachidonate induced aggregation and secretion. Leupeptin (100 ugs/ml) effectively blocked thrombin (0.2 u/ml) induced elevation of cytosolic calcium, but did not affect arachidonate induced elevation of platelet intracellular calcium levels. At a concentration of 100 ug/ml, Leupeptin effectively blocked thrombin (0.5u/ml) induced clot formation of platelet poor plasma, suggesting that it can exert its effect on thrombin by preventing fibrinogen degradation. Effective K_i for the competitive inhibition of thrombin induced hydrolysis of a chromogenic substrate, S2238, by Leupeptin was 2.4 uM. Leupeptin inhibition of platelet function was reversible by washing platelets free of the polypeptide. Results of our study demonstrate that Leupeptin inhibits thrombin induced platelet activation, probably by interfering with its proteolytic activity on the platelet surface membrane. However, inhibition of platelet surface membrane associated proteases did not prevent activation of platelets by other agonists.     

甲基黄酮醇胺盐酸盐对异丙肾上腺素正性频率作用的影响

本文比较了甲基黄酮醇胺盐酸盐(MFOA)、普萘洛尔(Pro)、维拉帕米(Ver)对异丙肾上腺素(Iso)所致兔离体右心房正性频率作用的影响。Pro使Iso累积浓度反应曲线平行右移,不抑制最大反应,属于典型的竞争性抑制剂,其pA2=8.43;MFOA和Ver使Iso量效曲线向下右移,抑制最大反应,为非竞争性拮抗。MFOA(2×10~(-5)M)和Ver(2×10~(-7)M)分别使最大反应下降19.28％和48.57％,其pD'2值分别为4.07±0.14、6.68±0.15。结果表明MFOA的作用不同于Pro,和Ver相似。     

Effect of intraperitoneally injected fluoride on plasma calcium in suckling and adult rats

Summary Sodium fluoride 10, 20, or 40 mg/kg body weight was given intraperitoneally to rats (6–11 days old and 90–95 days old). Blood analyses showed an initial increase in plasma fluoride concentration. The subsequent decrease in fluoride was paralleled by a decrease in total plasma calcium. These plasma concentrations were normal at blood collection 4 days after fluoride injection. The baby rats differed from the older rats in that their initial plasma calcium was higher and that the drop in plasma calcium concentration was less pronounced than in the old rats. A diet low in calcium and phosphate enhanced the effects of fluoride on total plasma calcium. The data indicate that the effect of large doses of fluoride on lowering the plasma calcium level is modified by the calcium intake.     

Distribution of cytochrome oxidase and parvalbumin in the primary visual cortex of the adult and neonate monkey,Callithrix jacchus

The anatomical distributions of the mitochondrial enzyme cytochrome oxidase (CO) and of the calcium binding protein parvalbumin (PV) were studied in the striate cortex of adult and neonate New World monkeys (Callithrix jacchus). In the adult marmoset, both proteins were found in laminar arrangements similar to those described for the macaque monkey, with prominent bands of PV-like immunoreactive (PV-LI) puncta in layers IV and IIIb, and fairly evenly distributed PV-LI nonpyramidal neurons. Furthermore, the pattern of CO activity in area 17 of the neonate marmoset was almost identical to the CO pattern described in neonate macaque and squirrel monkeys. It came, therefore, as a surprise to find that the adult pattern of PV-like immunoreactivity (PV-LI) in the marmoset striate cortex arises from a neonatal pattern strikingly different from that seen in any developmental stage of the macaque, or in any other mammal studied so far. In the deep layers IV through VI of the neonate marmoset, a large number of PV-LI neurons was stained in bandlike patterns, their number in layers IV and V exceeding the number of PV-LI neurons present in these layers of the adult marmoset area 17. Staining of layers IV and VI was restricted to area 17 and involved nonpyramidal cells and their exceeding the number of PV-LI neurons present in these layers of the adult marmoset area 17. Staining of layers IV and VI was restricted to area 17 and involved nonpyramidal cells and their processes. The stained band of layer V, in contrast, continued throughout most of the neocortex. In area 17, an estimated 10 to 20% of the stained cells in layer V exhibited pyramidal shapes. The findings show that the expression of PV by visual cortical cells occurs before birth and suggest that the comparatively early onset of PV expression is not dependent on the onset of textured vision. The exuberant number of stained cells in some layers, and particularly the staining of pyramidal cells, in the neonate marmoset, suggest that a considerable number of cells possesses the stainability for PV-LI only transiently, i.e., in the marmoset, these cells have a specific demand for parvalbumin during this phase of their development. © 1994 Wiley-Liss, Inc.