欢迎访问《解剖学报》官方网站！今天是
English
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
Decrease of the glial and fibrotic scars tissue formation by degrading fibrinogen after traumatic brain injury
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 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.
创伤性脑损伤 / 纤维瘢痕 / 胶质瘢痕 / 巴曲酶 / 免疫荧光 / 小鼠
Traumatic brain injury / Fibrotic scar / Glial scar / Batroxobin / Immunofluorescence / Mouse
［1］Mocchetti I, Wrathall JR. Neurotrophic factors in central nervous systemtrauma［J］. J Neurotrauma, 1995,12(5):853-870.
［2］Padilla CR. Syringomyelia after spinal cord injury［J］. Am Fam Physician, 1982,26(1):145-151.
［3］Vernon JD, Silver JR, Symon L. Post-traumatic syringomyelia: the results of surgery［J］. Paraplegia, 1983,21(1):37-46.
［4］Spencer T, Domeniconi M, Cao Z， et al. New roles for old proteins in adult CNS axonal regeneration［J］. Curr Opin Neurobiol, 2003,13(1):133-139.
［5］Asher RA, Morgenstern DA, Moon LD， et al. Chondroitin sulphate proteoglycans: inhibitory components of the glial scar［J］. Prog Brain Res, 2001,132:611-619.
［6］Moon LD, Asher RA, Rhodes KE， et al. Regeneration of CNS axons back to their target following treatment of adult rat brain with chondroitinase ABC［J］.Nat Neurosci,2001,4(5):465-466.
［7］Kawano H,Kimura-kuroda J,Komuta Y.Role of the lesion scar in the response to damage and repair of the central nervous system［J］.Cell tissue Res,2012,349(1):169-180.
［8］Yoshioka N,Kimura-Kuroda J,Saito T，et al. Small molecule inhibitor of type I transforming growth factor-β receptor kinase ameliorates the inhibitory milieu in injured brain and promotes regeneration of nigrostriatal dopaminergic axons.［J］.J Neurosci Res,2011,89(3):381-393.
［9］Raposo C,Schwartz M. Glial scar and immune cell involvement in tissue remodeling and repair following acute CNS injuries［J］.Glia,2014, 62(11):1895-1904.
［10］Soderblom C1, Luo X, Blumenthal E,et al. Perivascular fibroblast form the fibrotic scar after contusive spinal cord injury［J］.J Neurosci, 2013, 33 (34):13882-13887.
［11］Kawano H,Li HP,Sango K,et al.Inhibition of collagen synthesis overrides the age-related failure of regeneration of nigrostriatal dopaminergic axons［J］.J Neurosci Res,2005,80(2):191-202.
［12］Li HP,Homma A,Sango K, et al.Regeneration of nigrostriatal dopaminergic axons by degradation of chondroitin sulfate is accompanied by elimination of the fibrotic scar and glia limitans in the lesion site［J］. J Neurosci Res,2007,85(3):536-547.
［13］Mckeon RJ,Jurynec MJ,Buck CR.The chondroitin sulfate proteoglycans neurocan and phosphacan are expressed by reactive astrocytes in the chronic CNS glial scar［J］.J Neurosci,1999,19(24):10778-10788.
［14］Cua RC,Lau LW,Keough MB, et al.Overcoming neurite-inhibitory chondroitin sulfate proteoglycans in the astrocyte matrix［J］.Glia,2013,61(6):972-984.
［15］Bush TG,Puvanachandra N,Horner CH, et al.Leukocyte infiltration,neuronal degeneration,and neurite outgrowth after ablation of scar-forming,reactive astrocytes in adult transgenic mice［J］.Neuron,1999,23(2):297-308.
［16］Yoshika N,Hisanaga S,Kawano H.Suppression of fibrotic scar formation promotes axonal regeneration without disturbing blood-brain barrier repair and withdrawal of leukocytes after traumatic brain injury［J］.J Comp Neurol,2010,518(18): 3867-3881.
［17］Lord ST. Fibrinogen and fibrin:scaffold proteins in hemostasis［J］.Curr opin Hematol,2007,14(3)236-241.
［18］Grainger DJ,Wakefield L,Farndale RW, et al.Release and activation of platelet latent TGF-beta in blood clots during dissolution with plasmin［J］.Nat Med,1995, 1(9):932-937.
［19］Grainger DJ,Mosedale DE,Metcalfe JC.TGF-beta in blood:a complex problem.［J］. Cytokine Growth Factor Rev,2000,11(1-2):133-145.
［20］Asher RA,Morgenstern DA,Fidler PS, et al.Neurocan is upregulated in injured brain and in cytokine-treated astrocytes［J］.J Neurosci,2000,20(7):2427-2438.
［21］Komuta Y,Teng X,Yanagisawa H, et al.Expression of transforming growth factor-β receptors in meningeal fibroblasts of the injured mouse brain.［J］.Cell Mol Neurobiol,2010,30(1):101-111.
［22］Yang Y,Tian SJ,Wu L, et al.Fibrinogen depleting agent batroxobin has a beneficial effect on experimental autoimmune encephalomyeli-tis［J］.Cell Mol Neurobiol,2011,31(3):437-448.
［23］Li HP,Bai D,Gao J, et al.Immunohistochemical localization of dermatan sulfate in mouse brain after traumatic injury［J］.Acta Anatomica Sinica, 2012,43(2):166-170. (in Chinese)
李洪鹏，白丹，高杰,等.硫酸皮肤素在小鼠脑外伤后的经时表达 ［J］.解剖学报,2012,43(2): 166-170.
［24］Faulkner JR1, Herrmann JE, Woo MJ, et al. Reactive astrocytes protect tissue and preserve function after spinal cord injury［J］.J Neurosci,2004,24(9):2143-2155.
［25］Myer DJ, Gurkoff GG, Lee SM, et al. Essential protective roles of reactive astrocytes in traumatic brain injury［J］.Brain,2006,129(Pt 10):2761-2772.
抑制TGF-β1/ Smad2信号转导通路对中枢神经损伤后神经再生的促进机制;TGF-β1/ Smad2信号转导通路与中枢神经损伤后纤维性瘢痕形成的相关研究
/
| 〈 |
|
〉 |
