Interactions between the putative calcium entry promotor Bay k 8644 and pressor responses produced by α-1 and α2-adrenocepter agonists in the pithed normotensive rat

Summary Interactions between the putative calcium entry promotor Bay k 8644 and both -₁ and ₁-adrenocepter mediated increases in diastolic pressure were studied in the pithed normotensive rat. The ₁-adrenocepter mediated pressor responses elicited by B-HT920, TL-99, DP-6,7-ADTN and B-HT958 were potentiated by Bay k 8644, reflected by a leftward shift and an increase in the maximum of the log dose-pressor respinse curves. The -₁-adrenocepter effects elicited by cirazoline, methoxamine, (–)-amidephrine, St 587, (–)-phenylephrine and Sgd 101/75 were less enhanced by Bay k 8644. Only a leftward shift of the dose-response curves was observed, which was most pronounced for (–)-phenylephrine and Sgd 101/75. The -₁ and ₂-adrenocepter pressor components of (–)-noradrenaline were similarly distinguished by Bay k 8644 as observed for the selective -₁ or ₂-adrenocepter agonists.Effects of Bay k 8644 on the increase in diastolic pressure mediated by B-HT 920, St 587 and cirazoline were also studied after pretreatment with the calcium entry blocker nifedipine. After additional pretreatment with nifedipine the potentiation by Bay k 8644 observed for B-HT 920 and St 587 was more pronounced. The presence of nifedipine had no effect on the interaction between Bay k 8644 and cirazoline.It is concluded that Bay k 8644 behaves as a mirror image of nifedipine. The observation that Bay k 8644 enhances ₂-adrenocepter mediated pressor effects more effectively than ₁-adrenocepter increases in diastolic pressure is in accordance with the hypothesis of the more pronounced calcium dependency of ₂-adrenocepter mediated pressor responses. The data obtained for ceptor mediated pressor responses. The data obtained for St 587 and (–)-phenylephrine are in apparent contradiction to the finding that the pressor responses to the former drug are more markedly inhibited by calcium entry blockade than those of the latter. It is suggested that St 587 employs calcium channels which are already maximally modulated and that (–)-phenylephrine makes use of calcium channels which are in a rather inactive state. The hypothesis is put forward that the intrinsic activity of ₂-adrenocepter agonists reflects their ability to bring calcium channels in an active state.     

Oral Administration of Ginger-Derived Lipid Nanoparticles and Dmt1 siRNA Potentiates the Effect of Dietary Iron Restriction and Mitigates Pre-Existing Iron Overload in Hamp KO Mice

Intestinal iron transport requires an iron importer (Dmt1) and an iron exporter (Fpn1). The hormone hepcidin regulates iron absorption by modulating Fpn1 protein levels on the basolateral surface of duodenal enterocytes. In the genetic, iron-loading disorder hereditary hemochromatosis (HH), hepcidin production is low and Fpn1 protein expression is elevated. High Fpn1-mediated iron export depletes intracellular iron, causing a paradoxical increase in Dmt1-mediated iron import. Increased activity of both transporters causes excessive iron absorption, thus initiating body iron loading. Logically then, silencing of intestinal Dmt1 or Fpn1 could be an effective therapeutic intervention in HH. It was previously established that Dmt1 knock down prevented iron-loading in weanling Hamp (encoding hepcidin) KO mice (modeling type 2B HH). Here, we tested the hypothesis that Dmt1 silencing combined with dietary iron restriction (which may be recommended for HH patients) will mitigate iron loading once already established. Accordingly, adult Hamp KO mice were switched to a low-iron (LFe) diet and (non-toxic) folic acid-coupled, ginger nanoparticle-derived lipid vectors (FA-GDLVs) were used to deliver negative-control (NC) or Dmt1 siRNA by oral, intragastric gavage daily for 21 days. The LFe diet reduced body iron burden, and experimental interventions potentiated iron losses. For example, Dmt1 siRNA treatment suppressed duodenal Dmt1 mRNA expression (by ~50%) and reduced serum and liver non-heme iron levels (by ~60% and >85%, respectively). Interestingly, some iron-related parameters were repressed similarly by FA-GDLVs carrying either siRNA, including ⁵⁹Fe (as FeCl₃) absorption (~20% lower), pancreatic non-heme iron (reduced by ~65%), and serum ferritin (decreased 40–50%). Ginger may thus contain bioactive lipids that also influence iron homeostasis. In conclusion, the combinatorial approach of FA-GDLV and Dmt1 siRNA treatment, with dietary iron restriction, mitigated pre-existing iron overload in a murine model of HH.     

牛磺酸对培养乳鼠心肌细胞缺氧坏死的保护作用

目的　观察牛磺酸 (Tau)对缺氧引起的心肌细胞坏死有无直接保护作用。方法　将培养心肌细胞分为正常对照、Tau对照缺氧 Tau和缺氧 4个组。测定培养细胞上清液中肌酸磷酸激酶 (CPK)活性 ,用原子吸收法测定细胞内钙、镁含量 ,Annexin V- FITC/ PI双染流式细胞术和台盼蓝染色镜检检测坏死细胞。结果　 Tau可使缺氧时培养细胞上清液中CPK活性降低 30 % ,坏死细胞减少大约 2 0 %。减少细胞内钙超载和镁丢失。结论　 Tau对缺氧导致的心肌细胞坏死有直接保护作用 ,此作用可能与减少细胞内钙超载和镁丢失有关。     

复合纳米人工骨的注射、凝固及其机械性能研究

目的 构建半水硫酸钙和纳米羟基磷灰石为主的复合人工骨材料并对其注射性能、凝固性能和机械强度的影响因素进行考察.方法 测试不同液固比条件下复合材料的注射特性,25℃和37℃时分别测试不同液固比、不同二水硫酸钙促凝剂条件下的材料初、终凝时间和压缩强度,均与纯硫酸钙作对比.结果 液固比0.50以上时注射性能满意.无论何种液/固比,复合材料的凝固时间均较硫酸钙延长,37℃下的凝固时间较25℃下延长.一定范围内促凝剂用量过大或过小均使凝固时间延长.液固比越大或促凝剂用量越高,材料压缩强度越低.纳米磷灰石含量增大则材料强度降低.结论 合理掌握纳米磷灰石的比例,液固比和促凝剂的用量,是开发可注射纳米人工骨的关键.     

Single channel activity associated with the calcium dependent outward current inHelix pomatia

A recently improved version of the extracellular patch clamp technique (9, 13) was used to record currents from microscopic membrane areas of Helix neurons with predominant Ca²⁺ dependent outward currents. Current fluctuations in the patches consisted mainly of frequently interrupted, one-sided steps indicating discrete open-closed state changes of single channels with an ohmic conductance of approximately 19 pS. Frequency of occurrence of the elementary events compares with amplitudes of macroscopic currents during depolarizing voltage steps of varied amplitude. Average delays in appearance of the events vary in line with delayed time courses of the cell's outward current.     

羟苯磺酸钙对血液流变及血栓形成作用的实验研究

目的研究国产抗糖尿病微血管病变药物羟苯磺酸钙对血小板聚集、血栓形成、血液粘滞度、红细胞脆性以及出血时间等影响 ,为临床应用提供理论依据。方法与结果羟苯磺酸钙0.15、0.21和0.30g/kg,灌胃 ,每日1次 ,连续3d ,可明显抑制ADP和胶原引起的大鼠血小板聚集 ,其抑制率与对照组比较有显著性差异(P<0.01) ;明显降低大鼠血液粘滞度及改善大鼠红细胞中间脆性 ,其作用与对照组比较均有显著性差异(P<0.01)。羟苯磺酸钙0.15、0.30和0.60g/kg灌胃 ,一次给药 ,可明显抑制大鼠颈动静脉旁路血栓形成 ,其作用呈剂量依赖性 ,与对照组比较有显著性差异(P<0.01 ,P<0.05) ;对于小鼠 ,羟苯磺酸钙0.15、0.30和0.60g/kg 灌胃 ,一次给药 ,可明显对抗胶原诱导的血栓形成 ,其降低血栓引起的死亡和偏瘫率呈剂量依赖性 ,与对照组比较有显著性差异(P<0.01 ,P<0.05) ;能明显延长小鼠出血时间 ,与对照组比较有显著性差异(P<0.01)。结论羟苯磺酸钙对于糖尿病微血管病变相关的血液学指标有改善作用。     

The action of caffeine on inward barium current through voltage-dependent calcium channels in single rabbit ear artery cells

The effect of caffeine on inward current carried by barium ions through voltage-dependent calcium channels has been investigated in single rabbit ear artery cells using whole-cell voltage-clamp techniques. Caffeine (1 –30 mM) caused a rapid and reversible concentration-dependent blockade of barium current and a related compound, 3-isobutyl-1-methylxanthine (IBMX), was a more potent inhibitor of barium current. Caffeine-induced inhibition of barium current showed no voltage- or usedependence and caffeine did not alter the steady-state inactivation of barium current. The effect of caffeine was not blocked by extracellular or by intracellular ryanodine or inclusion of both 5 mM 1,2-bis(2-aminophenoxy)-ethane N,N,N,N,-tetraacetic acid (BAPTA) and 2 mM ethylene glycol-bis(-amino ethyl ether) N,N,N,N,-tetraacetic acid (EGTA) in the intracellular solution. Rolipram and M&B 22984, non-xanthine inhibitors of phosphodiesterase, did not diminish inward barium current. The data indicate that caffeine and IBMX block voltage-operated calcium channels and it is suggested that this is due to a direct interaction of methylxanthines with the calcium channel.     

Studies on subcutaneous fat necrosis of the newborn

Summary Biopsy specimens from the skin and subcutaneous fat tissue of four cases with neonatal subcutaneous fat necrosis were made and investigated by light and electron microscopy at 2, 4, and 6 weeks, and 5 months (Case 2) from the onset of the disease. Three stages of ultrastructural change of fat cells were observed. The evolution of crystal formation in the fat cells was seen and phagocytosis of crystals and fat droplets by macrophages and foreign-body giant cells was also noted. In the light microscope accumulation of calcium concretions in the spaces between and inside the fat cells was found. In the electron microscope we detected foci of highly electrondense granules, which were similar in distribution and structure to calcium salts stained with the von Kossa method. Changes in small and medium size blood vessels were observed.This work was written during a stay supported by Max-Planck-Gesellschaft from Dec. 1. 1973 to March 31. 1975 in the Max-Planck-Institut für Klinische und Physiologische Forschung (W.G. Kerckhoff-Institut), Bad Nauheim, West Germany (Director: Prof. Dr. Wolfgang Schaper)     

Homer 1b/c通过内质网功能调节由谷氨酸诱发的HT22细胞自噬

目的　研究Homer1b/c蛋白在谷氨酸诱发的细胞自噬中的作用及机制。方法　选用小鼠海马细胞系HT22细胞,通过500 ?mol/L谷氨酸处理建立细胞损伤模型。用siRNA慢病毒转染方式下调Homer1b/c表达和10 ?mol/LBAPTA-AM（1,2-双(2-氨基苯氧基)乙烷-N,N,N`,N`-四乙酸四乙酸甲酯,钙离子螯合剂）、10 mmol/L4-PBA（4-苯基丁酸,内质网应激抑制剂）分别抑制细胞内钙离子释放和内质网应激后,使用蛋白质印迹法检测Homer1b/c,自噬蛋白Beclin-1、微管相关蛋白轻链3（LC3）,以及内质网应激标志蛋白CHOP（人内质网应激相关蛋白）、GRP-78(葡萄糖调节蛋白78)的表达水平。每组实验均进行3次,采用独立样本t检验和单因素方差分析进行统计学分析。结果　谷氨酸处理HT22细胞12 h后,Beclin-1表达和LC3-Ⅱ/LC3-Ⅰ比值均升高（P<0.05）,下调Homer1b/c表达可降低Beclin-1表达和LC3-Ⅱ/LC3-Ⅰ比值的升高程度（P<0.05）。抑制细胞内钙离子释放和抑制内质网应激均能降低Beclin-1表达和LC3-Ⅱ/LC3-Ⅰ比值的升高程度（P<0.05）。然而在下调Homer1b/c表达后,抑制细胞内钙离子释放和抑制内质网应激未能进一步降低Beclin-1表达和LC3-Ⅱ/LC3-Ⅰ比值的升高程度（P<0.05）。结论　Homer1b/c能够调节谷氨酸诱导的自噬,其调节作用可能与内质网功能有关。     

Efficacy of cerebroprotective substances in the management of functional disorders induced by the cytotoxic brain oedema-producing substance hexachlorophene

Summary The hexachlorophene-induced cytotoxic brain oedema is used as experimental model of brain damage, suitable for testing cerebroprotective substances. It has clinical importance since many brain injuries are accompanied by an oedema. The primary target of the neurotoxin, hexachlorophene, is the neuronal cell membrane, but it also causes secondary effects including a disruption of myelin lamellae, increases in water and sodium content, decreases of potassium content, and vacuolation in the white matter. Rats received orally hexachlorophene 240 mg/kg a day for three weeks by liquid diet. The disruption of coordinative motor response, observed in a specially developed test, was used to characterise hexachlorophene-induced injuries in studies designed to evaluate the potential of cerebroprotective substances. Because of their membranotropic efficacy some nootropic substances with different modes of action were examined. The disturbance of coordinative motor response was restored significantly earlier than in spontaneous remission following administration of piracetam, pyritinol, methyl glucamine orotate, naftidrofuryl, and also under the influence of the calcium antagonists cinnarizine, flunarizine and nifedipine. These results support the therapeutic use of nootropic substances in the management of neurotoxic injuries and brain oedema.