非巴比妥类静脉全身麻醉药对大鼠皮层星形胶质细胞表达凝血酶敏感蛋白1及神经元突触数目影响的实验研究

【作者】 李俣；

【导师】 叶铁虎；

【作者基本信息】 北京协和医学院 ， 麻醉学， 2011， 博士

【摘要】 目的：通过体外细胞培养实验,研究依托咪酯、咪达唑仑及氯胺酮对星形胶质细胞表达凝血酶敏感蛋白的影响。并观察依托咪酯和氯胺酮对共培养神经元突触的影响。方法：1)分离纯化培养SD大鼠大脑皮层星形胶质细胞,并分别用依托咪酯、咪达唑仑及氯胺酮进行干预。药物设立不同浓度及不同时间组。利用RT-PCR和Western blotting技术分别检测星形胶质细胞表达TSP在基因和蛋白水平的变化；2)建立并优化星形胶质细胞和神经元共培养模型。用依托咪酯和氯胺酮进行处理,利用细胞免疫荧光化学法观察上述药物对神经元突触数目的影响。结果：1)依托咪酯可促进星形胶质细胞表达TSP-1,且具有浓度依赖性和时间依赖性。2)氯胺酮可减少星形胶质细胞分泌TSP-1,但与对照组无统计学差异。3)咪达唑仑对星形胶质细胞表达TSP-1无明显影响。4)依托咪酯可增加共培养神经元的突触数目；氯胺酮可减少共培养神经元的突触数目。两组之间具有统计学差异。结论：1.依托咪酯具有以促进星形胶质细胞TSP-1表达的作用。2.氯胺酮下调星形胶质细胞表达TSP-1,但与对照组相比无统计学差异；咪达唑仑对星形胶质细胞分泌TSP-1无明显影响。3.依托咪酯和氯胺酮可通过影响TSP-1的表达间接作用于神经元,分别对突触数量产生上调和下调的作用。更多还原

【Abstract】 Objective:To investigate the modulation of TSP-1 levels in astrocytes by etomidate, midazolam and ketamine and observe the influence on synapses number of co-cultured neurons by etomidate and ketamine.Method:Cortical astrocytes were cultured then treated with etomidate, midazolam and ketamine at different concentrations. The Sprague-Dawley rat was used as the primary cell source. The TSP-1 expression was detected through RT-PCR and Western blotting. We established co-culture model of neurons and cortical astrocytes and observed the numbers of synapses of co-culture neurons after being treated with etomidate and ketamine.Results:1) Etomidate promote the expression of TSP-1.2) Ketamine reduce the secretion of TSP-1, though there is no significant difference compared with the control group.3) Midazolam has no notable effect on the modulation of TSP-1.4) Etomidate increases the number of synapses of co-cultured neurons while ketamine reduces it.Conclusion:1) Etomidate promote the expression of TSP-1.2) Ketamine reduce the secretion of TSP-1.3) Midazolam has no obvious effect on the modulation of TSP-1. 4) Etomidate and ketamine has different influence on the synapses number of neurons.更多还原