补阳还五汤对全脑缺血模型大鼠再灌注不同时点皮质神经细胞钙超载的影响  

Effects of Buyang Huanwu Tang on Ca^(2+) Message of Global Cerebral Cortical Neurons in Rats with Ischemia and Reperfusion at Different Time Points

作　　者：关莉[1] 闫福曼[1] 张哓东[1] 苏文[1] 李小英[1] 周乐全[1] 罗荣敬[1] 

机构地区：[1]广州中医药大学,广东广州510405

出　　处：《安徽中医学院学报》2008年第4期45-49,共5页Journal of Anhui Traditional Chinese Medical College

摘　　要：目的:采用膜片钳技术和离子图像分析技术,同步观察补阳还五汤预干预对全脑缺血大鼠皮质神经细胞L型Ca2+通道及细胞内Ca2+浓度的影响,探讨Ca2+信号异常参与缺血性神经细胞损伤的机制及补阳还五汤抗脑缺血的分子机制,为临床应用本方防治缺血性脑病提供理论基础,并为中药复方研究探索新的思路和方法。方法:参照改良的Pulsinelli四血管闭塞法复制全脑缺血大鼠模型,缺血后大鼠分别在存活2,12,24,48及72 h后进行皮质神经细胞急性分离,负载荧光指示剂Fura-2/AM,用钙离子成像系统检测单个神经细胞内Ca2+浓度。单通道电流经EPC-9膜片钳放大器放大,用Pulsefit+Pulse采集至计算机。用分析软件TAC进行测量。结果:补阳还五汤预干预除再灌注12 h外,可显著降低其他各时点大脑皮质神经细胞内Ca2+浓度,对缺血再灌后两次Ca2+堆积均表现明显的抑制作用。补阳还五汤再灌注72 h,皮质神经细胞L型Ca2+通道平均开放时间显著降低,再灌注2 h,皮质神经细胞L型Ca2+通道开放概率显著降低。结论:降低缺血再灌注不同时点皮质神经细胞内Ca2+浓度及在缺血再灌注不同时点分别降低L型Ca2+通道开放概率和开放时间是补阳还五汤抗脑缺血的重要机制。补阳还五汤预干预对缺血性神经细胞损伤的预防和治疗有非常积极的意义。Objective： To apply the patch clamp and the ion-image analytical technique to simultaneously study the effects of Buyang Huanwu Tang on the Ca^2＋ L-channels and the intracellular Ca^2＋ concentrations of the ischemic rats＇ cerebral cortical neurons, intensively discuss the mechanism that the abnormity of the neuronal Ca^2＋ message joins in the ischemic neuronal injury and the molecular mechanism of cerebral anti-ischemia with BuYang Huanwu Tang, provide the theoretical basis for the prevention and treatment of the ischemic encephalopathy with this prescription, and explore new ideas and methods for the research on the complex prescription. Methods： Consulted the modified Pulsinelli＇s tetra-vessel occlusion to prepare the global cerebral ischemic rats. The rat cerebral cortical cell was acquired in the acute separation method. And the Ca^2＋ concentration of a single neuron was tested by the Ca^2＋ image-forming system. Records and analysis of the single-channel electricity, the recording single-channel current was amplified by the EPC-9 patch-clamping amplifier, and collected into the computer by Pulsefit ＋ Pulse. It was tested by the analytical software TAC. Fifty percent of the electrical amplitude at some recording voltage was considered as the transition to judge the channel＇s opening and closing. The acquired data was statistically processed by the software TACFIT, Results： The preintervention by Buyang Huanwu Tang had the definite effect on dropping the Ca^2＋ concentration of the global cerebral cortical neuron of rats with ischemia and reperfusion. Especially, the effect was more significant when the Ca^2＋ concentration reached the peaks （at 2 h and 24 h after reperfusion）. The preintervention by Buyang Huanwu Tang had the definite effect on dropping the opening probability of the Ca^2＋ L-channels of the rats＇ cerebral cortical neurons after repertfusion. Especially, the effect on drop was more significant when the probability reached the peak （at 2 h after reperfusion）

关 键 词：脑卒中 缺血再灌注 全脑皮质 神经细胞 钙离子信号 补阳还五汤 

分 类 号：R743.3[医药卫生—神经病学与精神病学] R285.5[医药卫生—临床医学]