马桑内酯对锥体神经元三磷酸腺苷敏感钾通道的作用
마상내지대추체신경원삼린산선감민감갑통도적작용
Effect of coriaria lactone on adenosine triphosphate-sensitive potassium channels in pyramidal neurons
  • 万方数据
    中国临床康复 중국림상강복
    CHINESE JOURNAL OF CLINICAL REHABILITATION
    2005年 45期 168-170 ,共3页

    邹晓毅%周华%周树舜

    추효의%주화%주수순

    神经元%钾通道%腺苷三磷酸%马桑内酯%膜片钳术 神經元%鉀通道%腺苷三燐痠%馬桑內酯%膜片鉗術
    신경원%갑통도%선감삼린산%마상내지%막편겸술
    背景:神经元异常放电的基础是细胞膜离子通道的激活与离子的跨膜运动.三磷酸腺苷敏感钾通道是将细胞电活动与代谢联系在一起的重要通道.三磷酸腺苷敏感钾通道是否参与癫痫的发病过程,马桑内酯是否具有调节三磷酸腺苷敏感钾通道的作用尚不清楚.目的:了解致痫剂马桑内酯对大鼠海马锥体神经细胞膜三磷酸腺苷敏感钾通道的影响及三磷酸腺苷敏感钾通道在癫痫发病中的作用.设计:随机对照实验.单位:四川大学华西医院神经内科和四川大学华西基础医学与法医学院生理学教研室.材料:实验于2000-05/12在泸州医学院完成.将Wistar乳鼠的培养的海马锥体神经元,随机分为正常对照组,四乙基胺组,二磷酸核苷组,马桑内酯组,电导与动力学组.方法:Wistar乳鼠在麻醉和无菌条件下分离出海马组织,接种、培养24 h后加入10 μmol/L的阿糖胞苷,选择培养7～10 d、生长良好、形态典型的锥体神经元进行膜片钳试验.正常对照组加入生理盐水;四乙基胺组加入5 mmol/L氯化四乙基胺;二磷酸核苷组先加入30 μmoL/L的二磷酸核苷,后加入0.5 mol/L的三磷酸腺苷;致痫组先加入1.0 mL/L的马桑内酯,后加入1 μmol/L的优降糖;对电导与动力学组,先调节钳制电压的大小,了解通道开放及通道形态,后加入马桑内酯.主要观察指标:①观察神经元三磷酸腺苷敏感钾通道的活动及形态.②观察不同钳制电压对通道活动的影响;了解二磷酸核苷、三磷酸腺苷和氯化四乙铵对通道的影响.③观察致痫剂马桑内酯对神经元细胞膜三磷酸腺苷敏感钾通道的激活作用及优降糖的影响.结果:①对称性高钾溶液条件下,通道的翻转电位接近0 mV.三磷酸腺苷敏感钾通道开放随着钳制电压绝对值的增大而增多,具有电压依赖性,该通道可被氯化四乙铵阻断.②其电流-电压(Ⅰ-Ⅴ)曲线可被直线拟合,电导值为(78.23±12.04)pS.③30μmol/L的二磷酸核苷可使通道开放增多,0.5 mol/L的三磷酸腺苷可抑制通道活动.④1.0mL/L的马桑内酯诱导通道开放数量明显增多,1μmol/L的优降糖可抑制通道活动.⑤通道开放时间,致痫神经元τ01为(1.754±0.060)ms,正常神经元为(1.733±0.046)ms,无显著性差异(n=25,t=0.147,P＞0.05);而τ02正常组为(2.441±0.265)ms,致痫组延长,为(10.446±0.579)ms(n=25,t=0.000,P＜0.01).结论:在马桑内酯诱导的癫痫发作中,三磷酸腺苷敏感钾通道开放的作用是降低动作电位频率、保护神经元,可能起一种负反馈调节作用.
    배경:신경원이상방전적기출시세포막리자통도적격활여리자적과막운동.삼린산선감민감갑통도시장세포전활동여대사련계재일기적중요통도.삼린산선감민감갑통도시부삼여전간적발병과정,마상내지시부구유조절삼린산선감민감갑통도적작용상불청초.목적:료해치간제마상내지대대서해마추체신경세포막삼린산선감민감갑통도적영향급삼린산선감민감갑통도재전간발병중적작용.설계:수궤대조실험.단위:사천대학화서의원신경내과화사천대학화서기출의학여법의학원생이학교연실.재료:실험우2000-05/12재로주의학원완성.장Wistar유서적배양적해마추체신경원,수궤분위정상대조조,사을기알조,이린산핵감조,마상내지조,전도여동역학조.방법:Wistar유서재마취화무균조건하분리출해마조직,접충、배양24 h후가입10 μmol/L적아당포감,선택배양7～10 d、생장량호、형태전형적추체신경원진행막편겸시험.정상대조조가입생리염수;사을기알조가입5 mmol/L록화사을기알;이린산핵감조선가입30 μmoL/L적이린산핵감,후가입0.5 mol/L적삼린산선감;치간조선가입1.0 mL/L적마상내지,후가입1 μmol/L적우강당;대전도여동역학조,선조절겸제전압적대소,료해통도개방급통도형태,후가입마상내지.주요관찰지표:①관찰신경원삼린산선감민감갑통도적활동급형태.②관찰불동겸제전압대통도활동적영향;료해이린산핵감、삼린산선감화록화사을안대통도적영향.③관찰치간제마상내지대신경원세포막삼린산선감민감갑통도적격활작용급우강당적영향.결과:①대칭성고갑용액조건하,통도적번전전위접근0 mV.삼린산선감민감갑통도개방수착겸제전압절대치적증대이증다,구유전압의뢰성,해통도가피록화사을안조단.②기전류-전압(Ⅰ-Ⅴ)곡선가피직선의합,전도치위(78.23±12.04)pS.③30μmol/L적이린산핵감가사통도개방증다,0.5 mol/L적삼린산선감가억제통도활동.④1.0mL/L적마상내지유도통도개방수량명현증다,1μmol/L적우강당가억제통도활동.⑤통도개방시간,치간신경원τ01위(1.754±0.060)ms,정상신경원위(1.733±0.046)ms,무현저성차이(n=25,t=0.147,P＞0.05);이τ02정상조위(2.441±0.265)ms,치간조연장,위(10.446±0.579)ms(n=25,t=0.000,P＜0.01).결론:재마상내지유도적전간발작중,삼린산선감민감갑통도개방적작용시강저동작전위빈솔、보호신경원,가능기일충부반궤조절작용.
    BACKGROUND: Abnormal neuronal discharge arose from the activation of cell membrane ion channels and transmembrane ion transport. The electric activity of the cells is associated with cell metabolism fundamentally through adenosine triphosphate (ATP)-sensitive potassium(KATP) channels.Currently the involvement of KATP channels in the pathogenesis of epilepsy and the regulation of KATP channels by coriaria lacton (EL) remain unknown.OBJETCIVE: To investigate the changes of cell membrane KATP channels in rat hippocampal neurons in response to CL as an epilepsy-inducing agent, and explore the role of KATP channels in the pathogenesis of epilepsy.DESIGN: Randomized controlled experiment.SETTING: Department of Neurology, West China Hospital Affiliated to Sichuan University, and Teaching and Research Section of Physiology,West China College of Preclinical Medicine and Forensic Medicine of Sichuan University.MATERIALS: This experiment was carried out at Luzhou Medical College between May and December 2000. Hippocampus pyramidal neurons were obtained from neonatal Wistar rats and randomized into normal control group, tetraethylammonium chloride (TEA) group, DNP group, CL group, and electric conductance and dynamics group.METHODS: The hippocampus of newborn Wistar rats was separated under aseptic condition and cultured for 24 hours prior to treatment with 10 μmol/L cytarabine for selective cell culture for 7-10 days. The cells in good growth exhibiting typical morphology of pyramidal neurons were then selected for patch-clamp experiment. The cells in the normal control group were treated with normal saline, which was replaced by 5 mmol/L TEA in TEA group, by 30 μmol/L DNP then 0.5 mol/L ATP in DNP group, and by 1.0 mL/L CL then 1 μmol/L glibenclamide in CL group. In electric conductance and dynamics group, the clamp voltage was firstly adjusted to investigate the channel opening before CL was added to the cells.MAIN OUTCOME MEASURES: ① Activity and curve of neuronal KATP channels; ② Effects of various clamp voltages on the channels activity and the effects of interventions with DNP, ATP and TEA; ③ Activation of neuronal membrane KATP channels induced by CL and the influence of glibenclamide.RESULTS: The reversal potential of the channels approximated 0 mV in homologous high-potassium solution. The opening of KATP channels increased along with the absolute value of the clamp voltage in a voltage-dependent manner, which was blocked by TEA. The electric current-voltage (Ⅰ-Ⅴ)curve could be fitted to a straight line with the electric conductance of (78.23±12.04) pS. Administration of 30 μmol/L DNP enhances the opening of the channels, which could be suppressed by 0.5 mol/L ATP.Addition of 1.0 mL/L CL to the cells caused obviously increased channel opening, which was suppressed by 1 μmol/L glibenclamide. The channel opening time was (1.754±0.060) ms for epileptic neuron τ01and (1.733±0.046) ms for normal neurons, showing no significant difference between them (n=25, t=0.147, P ＞ 0.05), but compared with the channel opening time of (2.441±0.265) ms for τ02 normal neurons, and duration was significantly prolonged in the epileptic neurons to reach (10.446±0.579)ms (n=25, t=0.000, P ＜ 0.01).CONCLUSION: The opening of KATP channels is responsible for reducing the action potential frequency for neuronal protection, which might be a negative feedback mechanism.
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