Flurazepam interaction with sodium and potassium channels in squid giant axon

External application of 0.05-1.0 mM flurazepam was found to partially block both sodium and potassium currents in voltage-clamped squid giant axons. At the same concentration the fractional block of the potassium current was found to be 3 times greater than that of the sodium current. In the presence of the drug the potassium current appeared to "inactivate', as flurazepam block became more profound during the course of the depolarization. The decay of the potassium current can be explained by a model in which flurazepam enters and blocks the potassium channels only after they have opened. Once bound in the potassium channel, removal of flurazepam from its binding site develops slowly (tau = 48 ms). Thus repetitive stimulation of the nerve produced a cumulative block. When applied inside the axon flurazepam was found to be 1.5 (n = 4) times more potent blocker of potassium channels than following external application. This result suggests that when applied externally, a neutral form of the drug diffuses across the membrane and blocks occurs from the inner end of the channel.     

硝普钠对豚鼠耳蜗外毛细胞全细胞钙电流作用的实验研究

目的　探讨一氧化氮供体———硝普钠 (sodiumnitroprusside ,SNP)对豚鼠单离耳蜗外毛细胞钙电流的影响及作用机制。方法　利用急性分离的豚鼠耳蜗外毛细胞 ,在全细胞膜片钳电压钳记录技术下 ,通过分离离子电流成分的方法 ,记录豚鼠耳蜗外毛细胞全细胞钙电流。结果　SNP对内向钙电流有抑制作用 ,在钳制电位为 -60mV ,刺激电压为 + 10mV的条件下 ,10mmol/L的SNP能抑制 (61 12± 1 99) %的内向钙电流 ( x±s ,n =5)。从 5～ 8个细胞得到的量效关系曲线中 ,其半数作用浓度为 1 9mmol/L ,最大抑制浓度为 10 0mmol/L ,斜率为 0 98。作用的机制为SNP可选择性地阻断外毛细胞膜上的L 型钙通道 (n =6)。结论　SNP作为一氧化氮的一种供体 ,能影响豚鼠耳蜗外毛细胞的生理功能 ,是通过阻断外毛细胞L 型钙通道电流的内流来实现的     

钾通道阻滞剂对大鼠支气管平滑肌细胞增殖的影响

目的探讨电压依赖性延迟整流钾通道(K_V)、Ca²⁺激活钾通道(K_Ca)和ATP敏感性钾通道(K_ATP)对大鼠支气管平滑肌细胞(BSMC)增殖的影响。 方法应用免疫细胞化学、MTT微量比色分析法及流式细胞术,观察K_V,K_Ca和K_ATP对培养中大鼠BSMC增殖的影响。结果K_V阻断剂4-氨基吡啶(4-AP)显著促进大鼠BSMC增殖细胞核抗原的表达,提高BSMC吸光度值,使S+G₂M期细胞数显著增多,并显著提高基础状态下BSMC内Ca²⁺浓度,而K_Ca阻断剂四乙铵(TEA)和K_ATP阻断剂格列本脲(Glib)均无此效应。 结论大鼠BSMC K_V活性的抑制,可提高细胞内Ca²⁺浓度,促进细胞的增殖,而K_Ca和K_ATP对BSMC的增殖均无明显影响。     

Puerarin blocks Na~+ current in rat ventricular myocytes

AIM: To study the effect of puerarin (Pue) on Na~+ channel in rat ventricular myocytes. METHODS: Whole-cell patch-clamp technique was applied on isolated cardiomyocytes from rats. RESULTS: Pue inhibited cardiac I_(Na) in a positive rate-dependent and dose-dependent manner, with an IC_(50) of 349 μmol/L. The kinetics of blockage of cardiac sodium channel by Pue resembled the ClassIa/Ic of antiarrhythmic agents. Pue 300 μmol/L did not alter the shape of the I-V curve of I_(Na), but markedly shifted the steady-state inactivation curve of I_(Na) towards more negative potential by 15.9 mV, and postponed the recovery of I_(Na) inactivation state from (21.9±1.6) ms to (54.4±3.4) ms (P<0.01 ). It demonstrated that the steady state of inactivation was affected by Pue significantly. CONCLUSION: Pue protected ventricular myocytes against cardiac damage and arrhythmias by inhibiting recovery from inactivation of cardiac Na~+ channels.     

Veratridine modifies the TTX-resistant Na channels in rat vagal afferent neurons

A number of neurotoxins from venoms of invertebrates and plants are ligands for voltage-gated Na⁺ channels and are useful tools for studying Na⁺ channel function and structure. Using whole-cell recordings from vagal afferent nodose neurons, we studied neurotoxins that target Na⁺ channels. We asked whether Ts3 (an α-scorpion toxin) and/or veratridine (a lipid-soluble toxin), could modify the TTX-resistant Na⁺ current generated by vagal afferent nodose neurons. Nodose TTX-resistant current was not affected by Ts3, whereas Ts3 slowed inactivation of the current generated by TTX-sensitive current component. We found that veratridine inhibited the TTX-resistant Na⁺ currents on rat nodose neurons. Interestingly, veratridine-modified Na⁺ channels developed a persistent current that accounted for the large tail current observed. We propose that veratridine modifies TTX-resistant Na⁺ channels through a mechanism distinct from its actions on other voltage-gated Na⁺ channels.     

N-甲基-D-天门冬氨酸受体及其调控的钙内流在大鼠感染性脑损伤中的变化

为探讨Ｎ－甲基－Ｄ－天门冬氨酸（ＮＭＤＡ）受体及其调控的钙内流在感染性脑损伤发病机制中的作用，采用Ｆｕｒａ－２／Ａｍ荧光法检测大鼠大脑皮质突触体内游离钙浓度（［Ｃａ＋＋］ｉ）。结果表明，注射百日咳杆菌组的［Ｃａ＋＋］ｉ较对照组显著增加（Ｐ均＜０．０１），并与注菌时间呈正相关关系（４小时ｖｓ０．５小时和２４小时ｖｓ４小时，Ｐ＜０．０５和＜０．０１）。ＮＭＤＡ受体的非竞争性拮抗剂ＭＫ－８０１预处理组（ＭＢＰ）的［Ｃａ＋＋］ｉ、脑含水量和伊文思蓝含量均较注菌组显著减少（Ｆ值分别为７２．２５、１６．８９、１４．３６，Ｐ＜０．０１和＜０．０５），并以２４小时［Ｃａ＋＋］ｉ减少最为明显（Ｐ＜０．０１）。提示ＮＭＤＡ受体调控的神经元内钙超载在感染性脑损伤、尤其在迟发性脑损伤的发病机制中起着重要作用。     

Cytochrome P450 epoxygenases as EDHF synthase(s)

The metabolism of arachidonic acid by cytochrome P450 (CYP) epoxygenases generates epoxyeicosatrienoic acids (EETs) which affect numerous cellular process including Ca(2+) signaling and the activity of Ca(2+)-dependent K(+) channels. The expression of the CYP epoxygenase(s) that generate EETs in endothelial cells is not constitutive but is determined by a number of physical (fluid shear stress and cyclic stretch) and pharmacological stimuli which also affect responses attributed to an endothelium-derived hyperpolarizing factor (EDHF). This review summarizes the role played by EETs, and the enzymes that generate and metabolize them, in EDHF-mediated responses.     

陈红风治疗乳腺癌中的扶正观

介绍陈红风教授治疗乳腺癌的临床经验。认为乳腺癌的发病与气血亏虚,肝、脾、肾及冲任二脉功能失调有关;治疗时重视通过调摄冲任以扶正、调理脏腑以祛邪。并附病例1则。     

葛根素对慢性偏头痛小鼠三叉神经节细胞钠离子电流及动作电位的影响

目的 通过观察葛根素对硝酸甘油诱导的慢性偏头痛模型小鼠行为学、三叉神经节细胞钠离子电流抑制率和动作电位的影响,探讨其治疗慢性偏头痛的作用机制。方法 随机取6只雄性C57BL/6小鼠,处死后分离三叉神经节细胞,检测不同浓度的葛根素对电压依赖性钠离子通道电流的抑制率。建立硝酸甘油诱导的慢性偏头痛小鼠模型,给予药物干预后,qRT-PCR检测三叉神经节细胞神经元电压门控钠通道基因（voltage-gated sodium channel α1-subunit gene,SCN1A）mRNA表达;用膜片钳记录各组小鼠三叉神经节细胞的动作电位参数。结果 葛根素对正常小鼠三叉神经节细胞的电压门控性钠离子通道抑制率的半数抑制浓度（half inhibitory concentration,IC₅₀）为29.96 μmol/L。与对照组比较,模型组小鼠0～30、30～60、60～90、90～120 min各时间段抓头次数及2 h内抓头次数总和显著增加（P＜0.05、0.001）,三叉神经节细胞中SCN1A mRNA表达显著升高（P＜0.001）,动作电位发放频率和静息膜电位显著升高（P＜0.001）。与模型组比较,葛根素（50、100、200 mg/kg）组30～60 min抓头次数和2 h内抓头次数总和显著减少（P＜0.05、0.01）,葛根素（200 mg/kg）组90～120 min抓头次数显著减少（P＜0.001）;葛根素（100、200 mg/kg）组小鼠三叉神经节细胞中SCN1A mRNA表达显著降低（P＜0.001）;葛根素（50、100、200 mg/kg）组小鼠三叉神经节细胞动作电位发放频率和静息膜电位显著下降（P＜0.001）。结论 葛根素通过阻断三叉神经节细胞钠离子电流和SCN1A mRNA表达,降低动作电位静息膜电位和发放频率,减少慢性偏头痛发作,有望成为临床治疗慢性偏头痛的辅助治疗药物。