Pinacidil reduces neuronal apoptosis following cerebral ischemia-reperfusion in rats through both mitochondrial and death-receptor signal pathways

Objective To investigate effect of pinacidil, an ATP sensitive potassium channel （KATP） opener, on the neuronal apoptosis and its signaling transduction mechanism following focal cerebral ischemia-reperfusion in rats. Methods One hundred male Wistar rats were randomly divided into four groups： A, sham-operated group; B, ischemia-reperfusion group; C, KATe opener treatment group; and D, KATe opener and blocker treatment group. The middle cerebral artery occlusion （MCAO） model was established by using the intraluminal suture occlusion method, neuronal apoptosis was determined by TUNEL staining, and expressions of caspase-8, caspase-9 and caspase-3 mRNA were detected by in situ hybridization. Results （1） The numbers of apoptotic neurons at 12 h, 24 h, 48 h, and 72 h were significantly less in group C than in groups B and D （P 〈 0.01 or P 〈 0.05）; and there was no difference between groups B and D at all time points （P 〉 0.05）. （2） The expressions of caspase-3 mRNA and caspase-8 mRNA at all times and the expressions of caspase-9 mRNA at 12 h, 24 h, 48 h, 72 h were significantly lower in group C than in groups B and D （P 〈 0.01 or P 〈 0.05）; and there were no differences between groups B and D at all time points （P 〉 0.05）. Conclusions KATP opener can significantly decrease the neuronal apoptosis and the expressions of caspase-3, caspase-8 and caspase-9 mRNAs following cerebral ischemiareperfusion. The neuronal apoptosis may be decreased by the inhibition of both mitochondrial and death-receptor signal pathways.

钾通道启开剂Pinacidil的硫脲／氰胍类似物的合成和降压活性

从吡啶基硫脲类化合物衍生、开发的Ｐｉｎａｃｉｄｉｌ是钾通道启开剂的代表药物之一。为寻找具钾通道启开作用的新型抗高血压药物，本文以Ｐｉｎａｃｉｄｉｌ类药物构效关系为依据，设计并合成了其４个硫脲（ＴＣ０１～ＴＣ０４）及８个氰胍类似物（ＰＣ０１～ＰＣ０８）。所有新化合物的结构均由元素分析，ＩＲ，ＭＳ和１￣ＨＮＭＲ证实。初步体内外药理试验表明：所合成的化合物均有不同程度的血管扩张及降压活性，其中以化合物ＰＣ０８的作用最强。

Pinacidil对低氧性肺动脉高压大鼠肺动脉压和血浆内皮素-1的影响

目的 :探讨钾通道开放剂pinacidil对低氧性肺动脉高压 (HPH)及对血浆内皮素 - 1(ET - 1)含量的影响。方法 :将 4 5只雄性Wistar大鼠分为 3组 :①对照组 ;②低氧组 ,每天低氧 8h ,共 4周 ;③治疗组 ,每天低氧前半小时腹腔注射pinacidil3mg/kg ,共 4周 ,4周后观察 3组平均肺动脉压 (mPAP) ,右心室肥厚指标、血浆中ET - 1的水平。结果 :①低氧组大鼠mPAP明显高于对照组。右心室肥厚显著 ,血浆中ET - 1含量明显高于对照组 ;②治疗组mPAP明显低于低氧组 ,右心室肥厚轻于低氧组 ,血浆中ET - 1含量明显低于低氧组。结论 :pinacidil能有效降低HPH中肺动脉压力、阻抑右心室肥厚 ,并影响血浆中ET - 1的水平。

Pinacidil的硫脲/氰胍类似物降压活性的定量构效关系研究

以ＰＯＬＹＧＥＮ软件包中的ＣＨＡＲＭｍ程序和集团坐标轮换法对合成的１５个ｐｉｎａｃｉｄｉｌ类似物及副产物的结构进行计算机分子模拟，分别以ＭＭ２法和ＣＤＮＯ／２法计算了上述化合物的１１个分子力学和１３个量子化学结构参数，将所得的结构参数与化合物的血管舒张活性进行相关分析，以逐步回归分析建立了两个相关性较好的方程。ｌｇ１／ＩＣ５０（１）＝－０．１２５９＋１．７１１ＤＭ＋０．０９３０ＰＳ（ｎ＝１６，Ｒ＝０．７４４４；Ｓ＝１．３１２１，Ｆ＝８．０７７１＞Ｆ２，１３（α≤０．０１）＝６．７０）；ｌｇ１／ＩＣ５０（２）＝１．２１１４－２８．７８４０ＱＮ（１）＋０．１２６５ＤＭ（ｎ＝１６，Ｒ＝０．７６５８，Ｓ＝０．４６１６，Ｆ＝９．２１８２＞Ｆ２，１３（α≤０．０１）＝６．７０）。结果表明，化合物的血管扩张活性随分子偶极矩（ＤＭ）、分子的溶剂可及疏水性表面积与亲水性表面积的比值（ＰＳ）、吡啶基氮原子电荷的绝对值［ＱＮ（１）］的增大而增大，化合物的血管扩张活性主要取决于分子的电荷分布和分子的疏水性。

模拟缺血-再灌液对窦房结细胞膜自发性动作电位的影响及Pinacidil的干预作用

目的 探讨模拟缺血 再灌液对窦房结细胞动作电位的影响及KKTP通道开放剂的干预效果。方法 取培养 2d的乳鼠窦房结细胞进行实验 ,随机分为对照组、模拟缺血 再灌注组 (I/R)、KATP通道开放剂Pinacidil干预组 (P +I/R)及KATP通道阻断剂 5 HD干预组 (5 HD +P +I/R及 5 HD +I/R)。采用全细胞膜片钳技术记录窦房结细胞动作电位 ,测定最大舒张电位 (MDP)、0期去极化速度 (UV)、超射值 (APO)、动作电位周期 (IBI)及 5 0 %复极时程 (APD50 )的变化。结果 ①与对照组比较 ,I/R组MDP(- 5 5 .2± 4 .5 )mV明显下降 (P <0 .0 5 ) ,APD50 (6 4.3± 3.8ms)明显缩短 (P <0 .0 1) ,而UV (3.8± 0 .5 )V/s及APO (14 .2±2 .0 )mV则明显减小 (P <0 .0 1) ,但IBI无明显改变。②与I/R组比较 ,P +I/R组的MDP及APO明显增大 ,IBI明显延长、UV显著加快 ,但APD50 进一步缩短。③ 5 HD不断阻断Pinacidil对模拟缺血 再灌液窦房结细胞MDP、IBI和APD50 的影响 ,但能使Pinacidil引起的UV及APO改变发生逆转。结论 Pinacidil可能通过开放不同种类KATP通道对模拟缺血 再灌液窦房结细胞动作电位产生影响 ,但这种影响能否为细胞提供保护有待进步研究。

钾通道开放剂Pinacidil及其类似物的合成(英文)

不同烷基取代的吡啶氰胍类化合物均具一定降压活性,Pinacidil是这类药物的代表,为一新型的降压药物。我们设计并合成了10个3-吡啶取代的类似物(均未见文献报道),并通过元素分析,MS,~1HNMR和IR确定了它们的结构。初步的药理试验表明,大部分化合物均有一定的降压活性,深入的药理工作正在进行之中。

不同浓度Pinacidil对兔心室肌细胞电生理特性的影响

目的:通过观察不同浓度的ATP敏感性钾通道(KATP)开放剂Pinacidil对家兔心室肌细胞电生理特性的影响,从而间接了解它不同程度的激活是否引起电活动的不均一性。方法:取健康新西兰大耳白家兔25只,体质量1.25～1.75kg,肝素处理后按改良的Taniguchi法分离单个心室肌细胞。分离出的单个细胞在12h内进行记录。用0.1mmol/LCdCI2阻断钙电流,分别用不同浓度的特异性开放剂Pinaci-dil干预,用全细胞膜片钳技术记录动作电位并测量相关参数。全细胞膜片钳记录按Hamill法进行。实验记录到的各项指标均进行统计学处理。结果:分别用3、10、30、100、300μmol/L的Pinacidil灌注,使细胞的复极化50%(APD50)和复极化90%(APD90)明显缩短,且随剂量增大缩短增加(r=-0.967,P<0.01)。对动作电位最大上升速度(Vmax)也有一定程度的影响,但无相关性(r=0.106,P<0.01)。不同浓度的Pinacidil对动作电位幅度(APA)几乎没有影响(r=0.038,P<0.01)。结论:不同浓度Pinacidil均能使APD缩短,其量效关系提示对KATP的激活程度不同,KATP可能使代谢状态不同的细胞之间产生电不均一性。

Protective Effects of Hyperpolarizing Cardioplegia with Pinacidil on Myocardium in Rats

Whether the ATP sensitive potassium channel opener pinacidil can provide myocardial protective effects in prolonged isolated global ischemic rat heart was investigated. On modified isolated rat working heart model, 40 hearts were divided into four groups randomly: Hyperpolarized arrest H K solution containing pinacidil (50 μmol/L) (P1 and P2) and depolarized arrest St. Thomas' solution (S1 and S2) subjected to 15 ℃ hypothermia, 60 min (P1 and S1) or 120 min (P1 and S2) of ischemia and 30 min reperfusion. The experimental indices included cardioplegic efficiency, cardiac function, coronary blood flow, myocardial enzyme release, myocardial water and ATP content. Hyperpolarized arrest provided significantly better recovery of cardiac function than depolarized arrest. Postischemic coronary flow and myocardial ATP content were higher. The arrest time of electro mechanical activities were longer than depolarized arrest. There were no differences among the groups in myocardial water contents. The hyperpolarized arrest solution containing pinacidil can provide a marked myocardial protective effect during prolonged hypothermic myocardial ischemia.

一种新的血管扩张剂—Pinacidil

pinacidil(P1134)是一种新型的血管扩张剂。由于它新的作用机制和作为一种新型的抗高血压药物而引人注意.1987年在美国印第安纳波里斯市举行了关于 Pinaci-dil 的专题座谈会。会议的论文专集收载了 Pinacidil 的基础研究及临床药理。

Myocardial protection of immature rabbits with an ATP-sensitive K^+ channel opener pinacidil

Objective To investigate the effectiveness of pinacidil,an opener of ATP-sensitive K+ channels,in protecting the myocardium of immature rabbit hearts from ischemic reperfusion injury.Methods Rabbit hearts underwent 30 min of global normothermic ischemia followed by 30 min of reperfusion on the modified Langendorff apparatus.Fifty-two isolated hearts of 3 - 4 week-old immature rabbits were divided into 4 groups randomly.During ischemia,3 different cardioplegic solutions were administered intermittently by infusion every 15 min(20-25 mi each time in all groups).Group 1:control group(n = 13);group 2:Krebs-Henseleit(K-H)solution with potassium(16 mmol/L)(n = 1:3);group 3:K-H solution with potassium(16 mmol/L)and pinacidil(50 μmol/L)(n = 13);group 4:K-H solution with potassium(16 mmol/L),pinacidil(50 μmol/L)and glibenclamide(10 μmol/L)(n = 13).The pre-ischemic and post-ischemic myocardial functions were assessed by the percentage recovery of the left ventricular developed pressure(LVDP);the left ventricular end-diastolic pressure(LVEDP);both the Positive peak and negative peaks of the first derivative of the left ventricular pressures(± dp/dtmax);coronary flow;the level of creatine kinase(CK),lactic dehydrogenase(LDH)and aspartate transcarbamoylase(AST)in coronary sinus venous effluent;and by myocardial ultrastructural changes.Results Before myocardial ischemia,there were no significant differences among the four groups in any of the parameters mentioned above.Post-ischemic recovery of LVDP,LVEDP,± dp/dtmax,coronary flow,the level of CK,LDH and AST,and myocardial ultrastructural changes were better in group 3 than those in the three other groups.Conclusions As a new and effective composition,pinacidil can significantly improve myocardial protection from cardioplegia for immature rabbit hearts.

Protective effects of pinacidil hyperpolarizing cardioplegia on myocardial ischemia reperfusion injury by mitochondrial KATP channels

Background Many studies have indicated that hyperpolarizing cardioplegia is responsible for myocardial preservation and researchers have suggested that the adenosine triphosphate-sensitive potassium channels （KATe） were the end effectors of cardio-protection. But whether mitochondrial KATe plays an important role in hyperpolarizing cardioplegia is not apparent. The present study investigated the effect of hyperpolarizing cardioplegia containing pinacidil （a nonselective KATe opener） on ischemia/repeffusion injury in rat hearts, especially the role of mitochondrial KATe in pinacidil hyperpolarizing cardioplegia. Methods Sprague-Dawley rat hearts were Langendorff-perfused for 20 minutes with Krebs-Henseleit buffer at 37℃ before equilibration. Cardiac arrest was then induced in different treatments： there was no arrest and ischemia in the normal group, the control group were arrested by clamping the aorta, depolarizing caidioplegia （St. Thomas solution containing 16 mmol/L KCI） and hyperpolarizing cardioplegia groups used St. Thomas solution containing 0.05 mmol/L pinacidil and 5 mmol/L KCI to induce cardiac arrest in group hyperkalemic and group pinacidil, in group hyperkalemic ＋ 5-hydroxydecanote （5HD） and Pinacidil ＋ 5HD, 5HD （0.1 retool/L） was added to the above two solutions to block mitochondria KATe channels. Global ischemia was then administrated for 40 minutes at 37℃, followed by 30 minutes of reperfusion. At the end of equilibration and reperfusion, hemodynamics, ultrastructure, and mitochondrial function were measured. Results In the control group, ischemia/reperfusion decreased the left ventricular developed pressure, heart rate, coronary flow, mitochondrial membrane potential, impaired mitochondrial respiratory function, increased reactive oxygen species and left ventricular end diastolic pressure. Damage to myocardial ultrastructure was also evident. Both depolarized arrest and especially hyperpolarized cardioplegia significantly reduced these lesions. 5HD partially blocked the beneficial effects of pinacidil cardioplegia but showing no effects on hyperkalemic arrest. Conclusions Pinacidil cardioplegia provides better cardioprotection with preservation of hemodynamics, ultrastructure, and mitochondrial function than traditional cardioplegia. The mitochondria KATe channels may play an important role in the protection mechanism.

Effects of pinacidil on proliferation of cultured rabbit airway smooth muscle cells induced by endothelin-1

It has been found that the potassium channel dysfunction of the membrane of airway smooth muscle cells (ASMCs) is closely associated with proliferation of ASMCs.~1 Preliminary research has demonstrated that pinacidil, an ATP sensitive potassium channel (K_(ATP)) opener, could play a remarkable role in the prevention and treatment of antigen induced bronchial asthma in guinea pigs.~2 (This study)was designed to investigate further the role and molecular mechanism of the proliferation of ASMCs: a chief pathological change of the nonacute phase of bronchial asthmatic episodes.

Differential effects of pinacidil,cromakalim,and NS 1619 on electrically evoked contractions in rat vas deferens

目的：本文研究新近研制的ＡＴＰ敏感性钾通道开放剂，吡那地尔（Ｐｉｎ）和ｃｒｏｍａｋａｌｉｍ（Ｃｒｏ），以及钙离子激活性钾通道开放剂ＮＳ１６１９对电场刺激所致大鼠输精管收缩的作用．方法：利用电场刺激（０．３Ｈｚ，１ｍｓ，６０Ｖ）反复性引致输精管单相性收缩．结果：Ｐｉｎ和Ｃｒｏ浓度依赖性减低电刺激收缩．格列本脲（Ｇｌｉ）而非ｃｈａｒｙｂｄｏｔｏｘｉｎ拮抗上述两药的舒张作用．Ｐｉｎ右移去甲肾上腺素的浓度－收缩曲线，同时降低最高收缩反应．Ｇｌｉ抵消Ｐｉｎ的作用．Ｃｈａｒｙｂｄｏｔｏｘｉｎ而非Ｇｌｉ减低ＮＳ１６１９的平滑肌舒张作用．结论：ＡＴＰ敏感性和钙离子激活性钾通道参与调节输精管平滑肌的收缩性．

Pinacidil suppression on 5-HT_3 receptor-mediated contraction of guinea pig ileum in vitro

目的：研究钾通道开放剂吡那地尔对５ＨＴ３受体介导的离体豚鼠回肠（ＧＰＩ）收缩反应的影响．方法：以等长换能器记录ＧＰＩ收缩反应．以［３Ｈ］ＧＲ６５６３０结合试验检测大鼠内嗅皮层５ＨＴ３受体的结合特性．结果：（１）２甲基５ＨＴ及５ＨＴ以剂量依赖方式引起ＧＰＩ的收缩；托品色创竞争性抑制此反应．（２）吡那地尔以剂量依赖方式抑制２甲基５ＨＴ和５ＨＴ引起的ＧＰＩ收缩，并增强托品色创或Ｂｅｎｅｓｅｔｒｏｎ对５ＨＴ诱发ＧＰＩ收缩的抑制作用，但不影响卡巴胆碱引起的ＧＰＩ收缩；吡那地尔对大鼠内嗅皮层５ＨＴ３受体与［３Ｈ］ＧＲ６５６３０的结合无影响．结论：吡那地尔可能通过激活突触前神经元ＡＴＰ敏感钾通道抑制由５ＨＴ３受体介导的ＧＰＩ收缩反应．

Pinacidil,a K_(atp) channel opener,identified as a novel agonist for TRPA1

The transient receptor potential Ankyrin 1(TRPA1) cation channel is activated by various pungent and irritant compounds,and it also mediates the perception of noxious cold.Identification of different agonists for this channel is important for understanding its activation mechanism.Therefore,a screen for novel TRPA1 agonists was performed using an agonist-induced calcium influx assay.Out of 90 compounds screened,pinacidil was identified as a novel agonist for this channel.Pinacidil is a known opener of the K atp channel,for which it has an EC50 value of 1-3 μmol/L.In comparison,the EC50 value of pinacidil for TRPA1 is relatively high(260 μmol/L).Recombinant HEK-TRPA1 cells did not respond to P1075,another K atp channel opener,suggesting that the effect of pinacidil on TRPA1 was highly specific.Further studies revealed that the agonist activity of pinacidil could be blocked by the TRP channel inhibitors,ruthenium red and HC-030031.Using glutathione(GSH) and site-specific mutagenesis,we demonstrated that pinacidil could activate TRPA1 by covalent modification of the critical amino acids C619,C639 and C663 in the N-terminus of TRPA1.

不同器官保存液对停搏供心的保护

背景:有研究已经证实应用人停搏供心进行心脏移植可以取得与非停搏供心移植相同的效果,但如何对停搏供心进行处理使之达到或接近非停搏供心移植的效果是一个新的课题。目的:采用新西兰大白兔离体停搏供心模型,探讨3种不同的器官保存液Celsior液、Pinacidile液和UW液对停搏供心的保护效果。设计、时间及地点:随机对照动物实验,于2005-03/2006-04在中南大学湘雅二医院胸心外科实验室心功能室完成。材料:选用新西兰大白兔24只,随机分成3组:UW液灌注组、Celsior液灌注组、Pinacidil液灌注组。方法:制作兔离体停搏供心的模型,当心热缺血10min后,分别应用4℃的UW液、Celsior液、Pinacidil液经主动脉根部冠状动脉开口灌注后4℃保存。4h后取出兔心固定在Langendorff灌流装置上复灌。主要观察指标:复灌后30,60,90,120min兔心脏血流动力学、2h内平均冠状动脉流量、心肌含水量及丙二醛含量。结果:复灌后120minPinacidil灌注组左室发展压、左室内压最大上升速率、左室内压最大下降速率均优于其余两组(P<0.05)。UW液灌注组复灌后120min心率高于另两组(P<0.05)。Pinacidile液灌注组和Celsior液灌注组心肌中含水量少于UW液灌注组,UW液灌注组心肌丙二醛含量高于Celsior液和Pinacidile液灌注组。复灌后120min内Pinacidile液灌注组心肌平均冠状动脉流量大于Celsior液和UW液灌注组。结论:Pinacidil液对停搏供心的保护优于UW液和Celsior液,表现为左室心功能及冠状动脉流量恢复得更好。

K^+通道开放剂对离体大鼠心肌缺血/再灌注损伤的保护作用

目的 观察三磷酸腺苷敏感性K+通道开放剂 (PCOs)Pinacidil对大鼠心肌缺血 /再灌注损伤的保护作用。方法 在改良Langendorff模型上 ,2 0只SD大鼠随机分为两组 ,分别用St.Thomas液 (对照组 )及经 10 μmol/LPinacidil强化的St.Thomas液 (Pinacidil组 )两种停跳液进行低温缺血 /再灌注大鼠离体心脏实验。离体心脏缺血90min ,再灌注 15min后观察缺血前、后心率 (HR)、左室收缩最高压 (LVSP)、左室舒张末压 (LVEDP)、最大压力变化速率 (±dp/dtmax)的变化 ,用RT PCR方法检测心肌组织eNOS mRNA和iNOS mRNA的表达变化 ,通过凝胶图象分析系统对RT PCR产物电泳条带进行密度分析。结果 Pinacidil组心功能指标明显好于对照组 ;eNOS明显升高 (P <0 .0 1) ,iNOS下降 (P <0 .0 5 )。结论 Pinacidil作为新的有效的停跳液成分可增强St.Thomas液对离体大鼠心肌缺血

钾通道开放剂——新型抗高血压药

Pinacidl(匹那地尔)为开放钾通道的新型抗高血压药的代表。其作用是使细胞膜钾通道开放,导致超极化,从而使平滑肌松弛,血管扩张,血压下降。其降血压效力与现有血管扩张药如肼笨哒嗪、哌唑嗪、硝苯吡啶等相当或略胜之。本品使用安全。主要不良反应为水肿。其它不良反应包括心悸、心率加速和暂时性 T 波变化。本品可使血脂降低,与噻嗪类利尿剂合用,可明显减轻其水肿副作用并增强其疗效。预期本品及其它钾通道开放剂今后将成为又一类新型抗高血压药。