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背景:既往有很多关于培养神经元或胶质细胞内钙的研究。然而关于高渗条件下共同观察神经元和胶质细胞内钙变化的研究目前较少。目的:探讨高渗刺激后大鼠下丘脑神经元和星形胶质细胞(AST)内钙的变化。设计:随机对照实验研究。单位:一所大学医院的神经外科和一所大学神经科学研究所的实验室。材料:实验在南方医科大学医院神经外科和解放军第四军医大学全军神经科学研究所完成。新生两三天的SD大鼠2只和孕18d大鼠胚胎6只,清洁级,由解放军第四军医大学实验动物中心提供。方法:体外联合培养的神经元和AST经钙荧光指示剂Fluo-3负载后,用激光扫描共聚焦显微镜测定高渗NaCl溶液影响前后的细胞内钙水平随时间变化情况。主要观察指标:联合培养的神经元和AST的形态观察及等渗和高渗状态下神经元和AST内Ca2+含量测定。结果:高渗刺激使培养神经元和AST内钙水平先升高后降低,最后维持在比高渗刺激前稍高的静息钙水平上。AST钙水平达到峰值的速度略快于神经元。结论:神经元和AST内Ca2+在渗透压信息传递过程起作用。
Background: There have been many previous studies on culturing neurons or glial intracellular calcium. However, there are few studies on the common observation of calcium changes in neurons and glial cells under hypertonic conditions. Objective: To investigate the changes of calcium in hypothalamic neurons and astrocytes (AST) after hypertonic stimulation in rats. Design: Randomized controlled trial. Unit: Neurosurgery at a university hospital and a laboratory at a university neuroscience institute. MATERIALS: Experiments were performed at the Department of Neurosurgery, Southern Medical University Hospital and the Institute of Neuroscience, Fourth Military Medical University of the People’s Liberation Army. Two newborn SD rats in two or three days and 6 pregnant embryos in 18d embryos were cleaned at the experimental animal center of the Fourth Military Medical University of Chinese PLA. Methods: The cultured neurons and AST in vitro were loaded with Fluo-3, a fluorescent probe of Fluo-3. Laser scanning confocal microscopy was used to measure the change of intracellular calcium level with time before and after the effect of hypertonic NaCl solution. MAIN OUTCOME MEASURES: Morphological observation of cultured neurons and AST and determination of Ca2 + content in neurons and AST under isotonic and hypertonic conditions. Results: Hypertonic stimulation increased the levels of calcium in cultured neurons and AST first and then decreased, and finally maintained at a slightly higher resting calcium level than before hypertonic stimulation. AST calcium levels peaked slightly faster than neurons. CONCLUSIONS: Ca2 + plays a role in the process of osmotic pressure transmission in neurons and ASTs.