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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。
Objective To explore the possibility of reducing the formation of glial scar and fiber scar after traumatic brain injury by degrading fibrinogen. Methods Eight-week-old Kunming mice were used to prepare mouse nigrostriatal pathway injury model according to Kawano’s method. The mice were anesthetized by intraperitoneal injection and fixed in a stereotaxic instrument. A rectangular notch was opened with a dental drill at a right rear 1.5 mm of the bregma and 6.0 mm vertically from the brain surface with a self-made 2.0 mm width blade. Then slowly pull out the blade to stop the suture. 24 Kunming mice were randomly divided into control group and experimental group. The experimental group was infused batroxobin 1h immediately after surgery for 3 days. On the 4th, 7th and 14th days after operation, coronal floating slice was taken at the level of brain. Col Ⅳ and GFAP antibodies were used to identify the expression of fiber scar and astrocyte in injured area. Double-labeled immunofluorescence was used to observe the formation of scar tissue at the injury site. Results On the fourth day after injury, Col Ⅳ was found in the injured area of the control group, and the peripheral membrane was formed by reactive astrocytes. On the 7th day, the scar formed in the center of the injury and the scar became more obvious on the 14th day. The same astrocyte surrounded; while the experimental group in the first 4 days of the injury site around the formation of astrocytes is difficult to form the realm of membrane, scar on the 7th and 14th days scarce, but the surrounding stars Surrounded by astrocytes. The results of double immunofluorescence showed that the control group had fibronectin (FN) at the center of injury, which resulted in the formation of fibrous scars. In the experimental group, the FN deposition decreased significantly after 7 days of injury and almost disappeared after 14 days. GFAP immunization Positive reaction-positive cells around. Conclusions Batroxobin injection attenuates the formation of fibrous scars and glial scar by degrading fibrinogen after brain injury.