星形胶质细胞钙调蛋白在损伤后胶质瘢痕形成中的作用

中枢神经系统损伤后的最后恢复大多数会形成胶质瘢痕[1]。胶质瘢痕形成的基础是星形胶质细胞的增生，其最重要的物质基础就是胶质纤维酸性蛋白（Glia fibrillary acidic protein ，GFAP ）。胶质瘢痕的形成对中枢神经系统损伤后神经结构和功能的

重建起到了严重的阻碍作用。因此，抑制胶质瘢痕的形成对中枢神经系统的损伤后修复起到了很好的促进作用。在前期实验中构建了大鼠星形胶质细胞损伤前后差异表达基因的cDNA 文库。在筛选差异表达的基因中,表达显著上调的基因包括包括热休克蛋白70基因，钙调蛋白基因等[2]。因此本实验主要探讨星形胶质细胞损伤后钙调蛋白对细胞增生的影响以及与胶质瘢痕形成之间的关系，为以后

作者介绍：顾香（1982-），女，硕士，

研究方向：神经损伤与修复。

通讯作者：陈阳美，男，教授，E-mail ：chenym1997@ 。

星形胶质细胞钙调蛋白在损伤后胶质

瘢痕形成中的作用

顾香1，张杰敏2，陈阳美1

（1．重庆医科大学附属第二医院神经内科；2．浙江衢州衢化医院神经内科，重庆400010）

【摘要】目的：探讨钙调蛋白对星形胶质细胞损伤后增生及胶质瘢痕形成的影响。方法：体外纯化培养星形胶质细胞，机械划痕法构建星形胶质细胞损伤模型，动态观察星形胶质细胞损伤前后钙离子浓度的变化，以及损伤后应用钙调蛋白抑制剂三氟拉嗪（Trifluoperazine dihydrochloride ，TFP ）处理，星形胶质细胞增生的主要标志物GFAP 表达的变化。采用激光共聚焦观察细胞内游离钙离子浓度动态变化，免疫组织化学染色方法检测星形胶质细胞胶质纤维酸性蛋白（Glia fibrillary acidic protein ，GFAP ）表达量的变化。

结果：星形胶质细胞损伤后细胞内钙离子浓度迅速上升，2min 后明显下降至接近基础水平。星形胶质细胞损伤后钙调蛋白抑制剂能够抑制星形胶质细胞GFAP 蛋白表达量的增加。结论：星形胶质细胞损伤后钙离子浓度明显变化，激活细胞内钙离子信号传导并参与星形胶质细胞损伤后的增生反应，而钙调蛋白抑制剂三氟拉嗪对星形胶质细胞损伤后的增生反应有显著的抑制作用。

【关键词】

星形胶质细胞；划痕损伤；增生；胶质瘢痕；钙调蛋白【中国图书分类法分类号】R338

【文献标识码】A

【收稿日期】2008-08-08

Effect of calmodulin on the formation of glial scar after the astrocyte injury

GU Xiang ，et al

（Department of Neurology ，the Second Affiliated Hospital ，Chongqing Medical University ）

【Abstract 】Objective ：To study the effect of calmodulin on reactive astrocytosis and formation of glial scar after astrocytes are injured.Methods ：Astrocytes were prepared from newborn rat cerebral cortex and evaluated the purity by immunostaining of glial fibrillary acidic protein （GFAP ）.The astrocyes injury model was made by means of surgical blade scraping.We observed the concentration of intracellular free calcium before and after injury in dynamic state .Then injured astrocytes were divided into two groups.The intervention group was added with suppressive agent of calmodulin-trifluoperazin dihydrochloride （TFP ）after injury ；the control group wae added into nothing.At different times after injury ，we performed immunocytochemical staining for glial fibrillary acidic protein （GFAP ）on each group and im-munocytochemical staining photographs were analyzed by image analysis system ，obtaining average optical densities.Results ：Concentra-tion of the tracellular free calcium increased quickly 2minutes after astrocytes were injured 。The immunocytochemical staining results in-dicated the control group and the intervention group had statistical significance.Conclusion ：After astrocytes were injured the intracellu-lar free calcium concentration obviously incredsed ，activated the intracellular Ca 2+signal pathway and participatal in the proliferation.TFP can obviously inhibit this proliferation reaction after the astrocytes are injured.【Key words 】Astrocyte ；Scraption ；Proliferation ；Glial scar ；Calmodulin

文章编号：0253-3626（2009）05-0625-04

临床研究