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背景:纳洛酮对各种类型的昏迷具有显著的促醒作用。一般认为纳洛酮的促醒作用与抑制内源性阿片肽(主要为β内啡肽)有关,但昏迷的程度并非一定与β内啡肽水平呈正相关。目的:在已证实促醒剂纳洛酮对皮质神经元具有直接兴奋作用的基础上,进一步研究阐明纳洛酮的兴奋皮质作用是否具有剂量依赖性。设计:以细胞为研究对象的单因素设计。单位:一所大学医院的神经内科,一所军医大学医院的神经内科。材料:实验在华中科技大学同济医学院实验中心完成。选取出生8～12d健康Wistar大鼠30只,体质量150～250g,雌雄不限。方法:实验在20～24℃的室温下进行。灌流槽置于倒置显微镜载物台上,选择表面光洁,胞体呈三角型或锥形且折光性强,有一个以上突起的神经元进行膜片钳实验。以急性分离Wistar大鼠额叶皮质锥体细胞为研究对象,采用膜片钳全细胞记录技术,观察不同剂量纳洛酮作用下,皮质神经元膜电位和自发性电活动频率变化的差异。主要观察指标:不同剂量纳洛酮的兴奋反应率、去极化幅度和自发电活动增加的比率。结果:急性分离的皮质神经元对100,50,10,1,0.1μmol/L等纳洛酮的兴奋反应率分别为83%,67%,86%,71%,33%,对应的去极化幅度分别为9.8,9.6,8.4,5.2和1.3mV,自发电活动增加的比率分别为587%,375%,291%,125%,69%。结论:
Background: Naloxone has a significant arousal effect on various types of coma. Naloxone is generally believed to have a potentiating effect on the inhibition of endogenous opioid peptides (mainly beta endorphin), but the degree of coma is not necessarily positively related to beta endorphin levels. OBJECTIVE: To further investigate whether naloxone, a stimulant of naloxone, has a direct stimulatory effect on cortical neurons in a dose-dependent manner. Design: Single-factor design with cells as the research object. Unit: Department of Neurology, a University Hospital, Department of Neurology, a Military Medical University Hospital. Materials: The experiment was completed at Tongji Medical College Experimental Center, Huazhong University of Science and Technology. Thirty healthy Wistar rats aged 8-12 days were selected and their body weight was 150-250 g. Methods: The experiment was carried out at room temperature of 20 ~ 24 ℃. The perfusion tank was placed on an inverted microscope stage, and the surface of the perfusion tank was selected to be smooth and smooth. The cell body was triangular or pyramidal and highly refractive. Neurons with more than one protrusion were subjected to patch-clamp experiments. Acute isolation of frontal cortex pyramidal cells of Wistar rats was used as object of study. The patch clamp whole cell recording technique was used to observe the changes of membrane potential and spontaneous electrical activity of cortical neurons under different doses of naloxone. MAIN OUTCOME MEASURES: The excitatory response rate of naloxone, the magnitude of depolarization, and the rate of increased spontaneous activity at different doses. Results: The excitatory response rates of acutely isolated cortical neurons to naloxone such as 100, 50, 10, 1 and 0.1 μmol / L were 83%, 67%, 86%, 71% and 33% 9.8, 9.6, 8.4, 5.2 and 1.3 mV, respectively, and the rates of self-generating activities increased by 587%, 375%, 291%, 125% and 69%, respectively. in conclusion: