中药川芎、当归有效成分对缺氧后复氧大鼠海马神经元钠、钾电流的作用

背景:中药红景天、人参、当归的有效成分具有抗缺氧损伤的作用。目的:从细胞电生理角度观察中药芎归滴丸对脑海马神经元的保护作用。设计:以细胞为观察对象,自身对照观察。单位:解放军第八十八医院和解放军军事医学科学院。材料:实验于2002-03/08在解放军军事医学科学院毒物药物研究所完成。选择新生清洁级Wistar大鼠1只,取其海马,将其分离培养成海马神经元细胞,然后选取9例海马神经元细胞,将其放入3个培养皿中培养,每个培养皿中3例神经元细胞,将其分为缺氧-复氧前后(对照组)、芎归滴丸(由解放军第八十八医院制剂室提供,主要成分为川芎和当归的挥发油)1×10-6mol/L组和芎归滴丸10×10-6mol/L组。方法:缺氧-复氧前后组分为两个亚组(即缺氧-复氧前、缺氧-复氧后),均不进行药物干预;芎归滴丸1×10-6mol/L组分为两个亚组(即芎归滴丸1.0×10-6mol/L 不缺氧组、芎归滴丸1×10-6mol/L 缺氧组);芎归滴丸10×10-6mol/L组分为两个亚组(即芎归滴丸10×10-6mol/L 不缺氧组、芎归滴丸10×10-6mol/L 缺氧组)。采用膜片箝全细胞记录方法检测缺氧-复氧前后体外培养的大鼠海马神经元的电压门控性Na 、K 电流。主要观察指标:体外培养的大鼠海马神经元的电压门控性Na 、K 电流幅度。结果:①海马神经元的电压门控性Na 电流幅度变化:缺氧-复氧后明显低于缺氧-复氧前犤(-3.8±1.0,-10.3±0.5)nA,t=3.49,P<0.01犦。②海马神经元的电压门控性K 电流幅度变化:缺氧-复氧后K 电流的激活阈值由-40mV变为-20mV,电流幅度变化差异无显著性意义。③芎归滴丸预处理后缺氧-复氧前后海马神经元的Na 、K 变化:缺氧-复氧前海马神经元ⅠNa、Ⅰk在阈值处的幅度很接近,差异无显著性意义。缺氧-复氧后芎归滴丸10×10-6mol/L组高于芎归滴丸1×10-6mol/L组和缺氧-复氧后犤(-58.5±1.9,-6.9±1.4,-3.8±1.0)nA,t=7.51,P<0.05犦。结论:芎归滴丸对海马神经元的缺氧损伤有抑制作用。

Effect of chuanxiong and danggui on Na+ and K+ currents of rat hippocampal neurons with hypoxia-reoxygenation

BACKGROUND: The active ingredients of Chinese herbs, such as hongjingtian (rhodiola root, Radix Rhodiolae), srenshen (ginseng, Radix Ginseng), and danggui (Chinese angelica root, Radix Angelicae Sinensis),were found to be effective to inhibit the hypoxia injury.OBJECTIVE: To observe the protecting effect of xionggui diwan (Dripping Pills) on the hippocampal neurons from hypoxia by whole-cell recording patch-clamp technique.DESIGN: An auto-control observation on cells.SETTING: The 88 Hospital of Chinese PLA and Academy of Military Medical Science of Chinese PLA.MATERIALS: The experiment was performed during March to August 2002 in the Institute of Toxicology and Pharmacology of Academy of Military Medical Sciences of Chinese PLA. The neonatal Wistar rats, clean degree, were used. The hippocampal neuron cells out of 9 rats were cultured and divided into the control groups of before and after hypoxia-oxygenation,and xionggui diwan 1 × 10-6 mol/L and 10 × 10-6 mol/L groups. (Xionggui diwan was provided by the 88 Hospital of Chinese PLA and its active ingredient was the volatile ingredient of chuanxiong and danggui.)METHODS: While hypoxia-reoxygenation groups were not given any medicine intervention as control, the treatment of xionggui diwan 1×10-6mol/L was subdivided into xionggui diwan 1 × 10-6 mol/L + non-hypoxia and xionggui diwan 1 × 10-6 mol/L + hypoxia groups, and the treatment of xionggui diwan 10 × 10-6 mol/L was subdivided into xionggui diwan 10 × 10-6 mol/L + non-hypoxia and xionggui diwan 10 × 10-6 mol/L + hypoxia groups. The patch-clamp technique was performed to determine the Na+ and K+ currents of rat hippocampal neurons cultured in vitro before and after hypoxia-reoxygenation.MAIN OUTCOME MEASURES: The Na+ and K+ current amplitude of rat hippocampal neurons cultured in vitro.RESULTS:① The Na+ current amplitude of hippocampal neurons: After hypoxia-reoxygenation, it was significantly lower than that before (-3.8±1.0, -10.3±0.5) nA, t=3.49, P ＜ 0.01]. ② The K+ current amplitude of hippocampal neurons: After hypoxia-reoxygenation, the activation thresholdof K+ current was changed from -40 mV to -20 mV.③The Na+ and K+ currents of hippocampal neurons before and after hypoxia-reoxygenation with xionggui diwan pretreatment: Before hypoxia-reoxygenation, the ampli tudes of Ⅰ Na and J K of hippocampal neurons at the threshold point were very similar, without significant difference. After hypoxia-reoxygenation,those amplitudes of xionggui diwan 10.0 × 10-6 mol/L group were higher than that of xiongguidiwan 1.0 × 10-6 mol/L group (-58.5±1.9, -6.9±1.4,-3.8±1.0) nA, t=7.51,P ＜ 0.05].CONCLUSION: It was suggested that xionggui diwan propect the function of hippocampal neurons from hypoxia.