上海交通大学学报（医学版）

›› 2010, Vol. 30 ›› Issue (4): 365-.

• 论著（基础研究） • 上一篇    下一篇

硫酸软骨素蛋白多糖Versican V1和V2亚型在大鼠损伤脊髓中的表达

顾文莉1, 吕合作2, 陆佩华2   

  1. 上海交通大学 1. 医学院第九人民医院检验科, 上海 200011, 2. 基础医学院神经生物学实验室, 上海 200025
  • 出版日期:2010-04-25 发布日期:2010-04-26
  • 作者简介:顾文莉（1972—）, 女, 副主任技师, 博士；电子信箱: guwenli8@yahoo.com.cn。
  • 基金资助:

    国家重点基础研究发展计划（“九七三”计划）（2003CB515302）；上海交通大学医学院附属第九人民医院第三届“优秀青年骨干”培养基金

Expression of Versican V1/V2 in injured spinal cord of rats

GU Wen-li1, LV He-zuo2, LU Pei-hua2   

  1. 1. Department of Clinical Laboratory, The Ninth People's Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200011, China；2. Department of Neurobiology, Basic Medical College, Shanghai Jiaotong University, Shanghai 200025, China
  • Online:2010-04-25 Published:2010-04-26
  • Supported by:

    National Basic Research Program of China, “973” Program, 2003CB515302；Foundation for The 3rd Excellent Young Core Members of  The Ninth People's Hospital, School of Medicine, Shanghai Jiaotong University

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摘要：

目的 探讨硫酸软骨素蛋白多糖Versican V1和V2亚型在成年大鼠损伤脊髓中的表达规律。方法 39只成年大鼠随机分为对照组（假手术组，n=9）和实验组（n=30）。利用纽约大学重物坠落装置建立实验组大鼠脊髓损伤模型。采用实时定量PCR、免疫组织化学法和免疫荧光法，分别观察大鼠脊髓损伤后Versican V1和V2亚型的表达变化。结果 实时定量PCR显示，与对照组比较，实验组Versican V1 mRNA的表达在脊髓损伤后4 h略有升高(P>0.05），损伤后1 d达到峰值(P<0.01)，其高表达可维持至损伤后7 d(P<0.01)；Versican V2 mRNA的表达升高较迟缓，脊髓损伤后1 d略有升高(P>0.05)，峰值出现在损伤后7 d(P<0.01)，然后迅速降低，损伤后14 d接近对照组水平(P>0.05）。免疫组织化学检测显示，脊髓损伤后3 d，包括损伤中心在内的10 mm全长脊髓都可观察到Versican表达明显增强；免疫荧光染色显示，高表达Versican的细胞为βⅢtubulin阳性神经元、GFAP阳性星型胶质细胞和MBP阳性少突胶质细胞。结论 Versican表达增强可能与脊髓损伤后形成抑制神经再生的微环境有关；消除其影响作用时，应考虑在不同时间采用不同方法处理表达模式不同的Versican 亚型。

关键词: 脊髓, 损伤, Versican, 大鼠

Abstract:

Objective To explore the expression patterns of Versican V1/V2 after injury of spinal cord in adult rats. Methods Thirty-nine adult rats were randomly divided into control group (sham-operation group, n=9) and experiment group (n=30), and rat models of spinal cord injury were established in experiment group using New York University impact injury device. The changes of Versican V1/V2 expression after spinal cord injury were observed by Real-time PCR, immunohistochemistry and immunofluorescent technique. Results Real-time PCR revealed that compared to control group, the expression of Versican V1 mRNA in experiment group slightly increased 4 h after injury (P>0.05), reached the peak 1 d after injury (P<0.01) and remained at elevated levels up to 7 d after injury (P<0.01). While the expression of Versican V2 mRNA increased a little slowly, slightly increased 1 d after injury (P>0.05) and peaked 7 d after injury (P<0.01), then diminished quickly, and returned to the level of control group 14 d after injury (P>0.05). Immunohistochemistry detection demonstrated that the expression of Versican significantly increased in injured spinal cord parenchyma 3 d after injury, and its staining was observed throughout the entire length of a 10 mm-long cord segment containing the injury epicenter. Immunofluorescent staining indicated that colocalization of Versican was found in the neurons with positive βⅢ-tubulin, astrocytes with positive GFAP and oligodendrocytes with positive MBP. Conclusion The increased expression of Versican may be involved in forming the environment of inhibiting axonal regeneration following injury. Optimization of strategies to enhance regeneration may need to target different Versican isoforms over an extended period following injury.

Key words: spinal cord, injury, Versican, rat