解剖学报

解剖学报 ›› 2016, Vol. ›› Issue (3): 303-308.doi: 10.16098/j.issn.0529-1356.2016.03.003

• 神经生物学 • 上一篇    下一篇

改良的体外中枢神经系统损伤后胶质瘢痕模型的建立

李奕 谈玲 林巍巍 陈雪 陈颖 潘静莹 王晓冬*   

  1. 江苏省南通大学医学院组织学与胚胎学系，江苏 南通 226001
  • 收稿日期:2015-08-20 修回日期:2015-10-23 出版日期:2016-06-06 发布日期:2016-06-06
  • 通讯作者: 王晓冬 E-mail:wxdzw@ntu.edu.cn
  • 基金资助:

    siRNA干扰EphB2抑制胶质疤痕形成及组织工程修复脊髓损伤

An improved glial scar model after central nervous system injury in vitro

LI Yi TAN Ling LIN Wei-wei CHEN Xue CHEN Ying PAN Jing-ying WANG Xiao-dong*   

  1. Department of Histology and Embryology, College of Medical Sciences, Nantong University, Jiangsu Nantong 226001, China
  • Received:2015-08-20 Revised:2015-10-23 Online:2016-06-06 Published:2016-06-06

摘要：

目的 建立胶质疤痕体外模型，观察其形态及细胞外基质的表达情况。方法 体外分别培养来自大脑皮层的星形胶质细胞和脑膜成纤维细胞，经胶质纤维酸性蛋白（GFAP）和纤连蛋白（FN）抗体免疫细胞化学染色鉴定后，将两种细胞混合共培养，2d后添加转化生长因子-β1（TGF-β1），以未添加TGF-β1为对照组；GFAP和FN免疫细胞化学染色观察胶质疤痕的形态；免疫细胞化学染色和Western blotting观察促红素肝细胞受体B2（EphB2）、Eph受体作用配体B2（ephrinB2）、神经蛋白聚糖（neurocan）和神经抗原2（NG2）表达情况。 结果 疤痕样细胞团簇结构主要是由星形胶质细胞和成纤维细胞共同组成；实验组EphB2、ephrinB2和neurocan、NG2表达量较对照组明显增加（P＜0.05）。 结论 体外混合培养星形胶质细胞与成纤维细胞并添加TGFβ1后成功建立体外中枢神经系统损伤后疤痕模型。

Abstract:

Objective To establish a scar model in vitro and observe the morphology and extracellular matrix of the scar. Methods Astrocytes and meningeal fibroblasts were cultured from the cerebral cortex in vitroand identified by glial fibrillary acidic protein (GFAP) and fibronectin(FN) immunofluorescence cell staining respectively. Astrocytes and meningeal fibroblasts were co-cultured and treated by TGF-β1 after 2 days. The group without TGF-β1 treating was used as the control. The morphology of scar was observed by GFAP and FN immunofluorescence cell staining. Immunofluorescence cell staining and Western blotting were used to examine the expression of EphB2, ephrinB2, neurocan and NG2 in the scar. Results The scar-like cell cluster was formed from astrocytes and fibroblasts. In the TGF-β1 group, the expressions of EphB2 and ephrinB2, neurocan and NG2 were significantly increased when compared with the control group (P<0.05). Conclusion A scar model after central nervous system injury has bean established by co-culturing astrocytes and fibroblasts treate by TGF-β1 in vitro.