上海交通大学学报（医学版）

›› 2012, Vol. 32 ›› Issue (12): 1567-.doi: 10.3969/j.issn.1674-8115.2012.12.011

• 论著（基础研究） • 上一篇    下一篇

损伤后脑膜成纤维细胞对星形胶质细胞活性及胶质纤维酸性蛋白表达的影响

李 奕1, 陈 雪1, 陈 颖1, 林巍巍1, 刘永华1, 朱俊杰1, 费 翔1, 刘宇西1, 王晓冬2   

  1. 1.南通大学医学院组织学与胚胎学教研室, 南通 226001； 2.南通大学附属医院神经外科, 南通 226001
  • 出版日期:2012-12-28 发布日期:2012-12-31
  • 作者简介:李 奕（1978—）, 女, 讲师, 硕士；电子信箱: ly1203@ntu.edu.cn。
  • 基金资助:

    国家自然科学基金（30870643）；江苏普通高校自然科学研究项目（11KJB310007）；江苏高校优势学科建设工程资助项目；南通大学校级自然科学基金（10Z055）

Effects of injured meninges fibroblasts on viability and expression of glial fibrillary acidic protein of astrocytes

LI Yi1, CHEN Xue1, CHEN Ying1, LIN Wei-wei1, LIU Yong-hua1, ZHU Jun-jie1, FEI Xiang1, LIU Yu-xi1, WANG Xiao-dong2   

  1. 1.Department of Histology and Embryology, Medical College, Nantong University, Nantong 226001, China；2.Department of Neurosurgery, Affiliated Hospital of Nantong University, Nantong 226001, China
  • Online:2012-12-28 Published:2012-12-31
  • Supported by:

    National Natural Science Foundation of China, 30870643；Jiangsu College Natural Science Research Foundation, 11KJB310007；Jiangsu College Excellent Discipline Construction Foundation；Nantong University Foundation, 10Z055

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摘要：

目的 体外观察损伤后脑膜成纤维细胞对星形胶质细胞活性及胶质纤维酸性蛋白（GFAP）表达的影响。方法 体外分别培养SD大鼠大脑皮层的星形胶质细胞和大脑脑膜的成纤维细胞，经GFAP和纤维粘连蛋白（FN）免疫荧光染色鉴定后，将损伤后成纤维细胞来源条件培养基与星形胶质细胞共培养，根据成纤维细胞损伤时间不同分为正常对照组（A组）、损伤4 h 组（B组）、损伤24 h组（C组）、损伤72 h组（D组），应用CCK8试剂检测星形胶质细胞的活性，通过GFAP免疫荧光染色观察星形胶质细胞GFAP表达。将损伤的成纤维细胞与星形胶质细胞接触共培养72 h，根据与成纤维细胞的距离，将星形胶质细胞分为近（J）组和远（Y）组，通过FN和GFAP双重免疫荧光细胞化学染色观察GFAP表达。结果 B组星形胶质细胞活性低于A、C、D组（P<0.05)；非接触共培养时，C组星形胶质细胞GFAP表达强度显著高于B、D组（P<0.05)；接触共培养时，J组星形胶质细胞的GFAP表达强度显著高于Y组（P<0.05)。结论 损伤后脑膜成纤维细胞能触发星形胶质细胞反应性胶质化。

关键词: 成纤维细胞, 星形胶质细胞, 细胞活性, 胶质纤维酸性蛋白

Abstract:

Objective To investigate the in vitro effects of injured meninges fibroblasts on viability and expression of glial fibrillary acidic protein (GFAP) of astrocytes. Methods Astrocytes from cerebra cortices and fibroblasts from meninges of SD rats were cultured respectively, and were identified by GFAP and fibronectin(FN) immunofluorescence staining. According to the terms of scratch injury of fibroblasts, astrocytes co-cultured with fibroblasts derived conditional medium were divided into normal control group (group A), 4-h injury group (group B), 24-h injury group (group C) and 72-h injury group (group D). The viability of astrocytes was determined by CCK8 assay, and the expression of GFAP of astrocytes was detected by GFAP immunofluorescence staining. At the same time, contact co-culture was conducted in injured fibroblasts and astrocytes for 72 h. The astrocytes were divided into short distance group (group J) and long distance group (group Y) according to the distance from fibroblasts, and the expression of GFAP was detected with FN and GFAP double immunofluorescence cytochemical staining. Results The viability of astrocytes in group B was significantly lower than that in group A, group C and group D (P<0.05). The expression of GFAP of astrocytes in group C was significantly higher than that in group B and group D (P<0.05). The expression of GFAP of astrocytes in group J was significantly higher than that in group Y (P<0.05). Conclusion Injured meninges fibroblasts are able to trigger reactive astrogliosis.

Key words: fibroblasts, astrocytes, viability, glial fibrillary acidic protein