降解纤维蛋白原可减少创伤性脑损伤后胶质瘢痕与纤维瘢痕的形成

doi:10.3969/j.issn.0529-1356.2014.06.001

摘要/Abstract

摘要：

目的探讨通过降解纤维蛋白原减少创伤性脑损伤后胶质瘢痕与纤维瘢痕的形成的可能性。方法选用8周龄昆明小鼠按照川野的方法制备小鼠黑质纹状体通路损伤模型。小鼠经腹腔注射麻醉后固定在脑立体定位仪上。在前囟点右后方1.5mm处用牙钻打开一长方形缺口，用自制宽度2.0mm的刀片从大脑表面垂直插入6.0mm。然后缓慢拔出刀片，止血缝合。24只昆明小鼠随机分成对照组与实验组。实验组于手术后1h立即注入巴曲酶注射液,连续3d。术后第4、7、14天取脑行水平位冠状浮游切片。应用胶原蛋白Ⅳ(ColⅣ)及GFAP抗体特异性识别损伤区域纤维瘢痕及星形胶质细胞的表达。应用双标免疫荧光法观察损伤部位的瘢痕组织形成。结果在伤后第4天，对照组的损伤部位出现ColⅣ 沉着，周边出现由反应性星形胶质细胞形成的境界膜，第7天后损伤中心形成纤维性瘢痕，第14天后瘢痕更明显，周边同样有星形胶质细胞包围；而实验组在第4天的损伤部位周边不易形成星形胶质细胞的境界膜，第7天及第14天纤维性瘢痕几乎不存在，但周边仍被星形胶质细胞所围绕。双重免疫荧光显示，对照组的损伤中心有纤维连接蛋白（FN）沉着，形成纤维性瘢痕；而实验组在损伤7d后FN沉着明显减少，14d后几乎消失；两组在损伤周边都有GFAP免疫阳性反应阳性细胞围绕。结论在脑损伤后，注入巴曲酶可通过降解纤维蛋白原减弱纤维性瘢痕及胶质瘢痕的形成。

关键词: 创伤性脑损伤, 纤维瘢痕, 胶质瘢痕, 巴曲酶, 免疫荧光, 小鼠

Abstract:

Objective To investigate probability of reducing formation of glial and fibrotic scar by degrading fibrinogen after traumatic brain injury. Methods The nigrostiatal dopaminergic pathway was unilaterally transected in 8-weeks-old Kunming mouse according to the method of Kawano et al. Adult male mice were anesthetized and transferred to a stereotaxic frame. A small oblong hole at the right rear of the bregma at 1.5mm and at a depth of 6.0 mm from the surface of the brain was made with a dental drill.. The blade was slowly pulled out, bleeding was stopped and the incision was sutured. Twenty four Kunming mice were randomly divided into control group and experimental group. For three consecutive days, experimental group mice were injected with batroxobin at 1 hour after operation. The mice brains were obtained at 4days, 7days and 14days after brain injury. Immunohistochemical localization of fibrotic scar and glial scar were examined by using the antibodies collagenⅣ(ColⅣ) and GFAP. The formation of scar tissue was observed by double immunofluorescent staining. Results After 4 days injury, the lesion site of the control group appeared ColⅣ deposition,which was surrounded by glial limitans formed by reactive astrocytes. Compared with the fibrotic scar of the lesion center 7days after injury, more obvious ColⅣ deposition was found at 14days than 7days after injury, and the lesion center was surrounded by the reactive astrocytes. However, the glial limitans were not found around the lesion center in the experimental group and fibrotic scar was almost disappeared, but the reactive astrocytes surrounded the lesion center. Double immunofluorescent staining demonstrated that there was FN immunoreactivity deposition in the lesion site of control groups forming fibrotic scar, while the experimental group reduced the deposition of FN significantly 7days after injury, and almost eliminated it at 14days after injury. Both groups had the GFAP positive cells surrounded the lesion site. Conclusion After traumatic brain injury, injection of batroxobin could decrease the formations of the glial scar and fibrotic scar by degrading fibrinogen.

Key words: Traumatic brain injury, Fibrotic scar, Glial scar, Batroxobin, Immunofluorescence, Mouse

白丹李丹刘囡裴丹李洪鹏*. 降解纤维蛋白原可减少创伤性脑损伤后胶质瘢痕与纤维瘢痕的形成[J]. 解剖学报, 2014, 45(6): 729-734.

BAI Dan LI Dan LIU Nan PEI Dan LI Hong-peng*. Decrease of the glial and fibrotic scars tissue formation by degrading fibrinogen after traumatic brain injury[J]. AAS, 2014, 45(6): 729-734.

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