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。
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.
引文
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