上海交通大学学报（医学版）

›› 2012, Vol. 32 ›› Issue (7): 881-.doi: 10.3969/j.issn.1674-8115.2012.07.013

• 论著（基础研究） • 上一篇    下一篇

鞘内注射经外源性BDNF活化的星形胶质细胞对正常大鼠痛觉的影响

曾路路1,2, 汪 静1,2, 张 昕1, 周全红2, 江 伟2, 杜冬萍1   

  1. 上海交通大学附属第六人民医院 1.疼痛科, 2.麻醉科, 上海 200233
  • 出版日期:2012-07-28 发布日期:2012-08-17
  • 通讯作者: 杜冬萍, 电子信箱: dudp@sjtu.edu.cn。
  • 作者简介:曾路路(1986—), 女, 硕士生；电子信箱: zenglu1986@126.com。

Effects of intrathecal injection of exogenous BDNF-activated astrocytes on pain sensation of normal rats

ZENG Lu-lu1,2, WANG Jing1,2, ZHANG Xin1, ZHOU Quan-hong2, JIANG Wei2, DU Dong-ping1   

  1. 1.Pain Management Center, 2.Department of Anesthesiology, the Sixth People´s Hospital, Shanghai Jiaotong University, Shanghai 200233, China
  • Online:2012-07-28 Published:2012-08-17

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摘要：

目的 探讨外源性脑源性神经营养因子（BDNF）对星形胶质细胞的活化作用，分析活化的星形胶质细胞鞘内注射与正常大鼠痛觉过敏的关系。方法 体外培养的原代星形胶质细胞以外源性BDNF（100 ng/mL）分别孵育0（基线）、15、60、120 min，Western blotting检测细胞内纤丝酸性蛋白（GFAP）的表达。24只雄性SD大鼠经鞘内置管后随机分为活化细胞注射组（鞘内注射经BDNF孵育15 min的星形胶质细胞）、阴性对照组（鞘内注射未经BDNF孵育的星形胶质细胞）、安慰剂注射组（鞘内注射PBS）和空白对照组(未行鞘内注射)，每组6只；分别于单次注射前和注射后0.5、2、4和8 h时间点，von Frey纤维丝法测定各组大鼠50%机械痛缩足阈值（50%PWT）。结果 经BDNF孵育15、60和120 min的星形胶质细胞内GFAP蛋白表达均显著高于基线值（P<0.01）。活化细胞注射组大鼠注射后各时间点的50%PWT均较注射前显著降低(P<0.01)；阴性对照组大鼠50%PWT仅在注射后30 min时间点呈现一过性下降，与注射前比较差异有统计学意义(P<0.01）；安慰剂注射组和空白对照组大鼠注射前和注射后各时间点50%PWT比较，差异均无统计学意义（P>0.05）。结论 外源性BDNF可活化体外培养的星形胶质细胞，鞘内注射经外源性BDNF活化的星形胶质细胞可直接诱发正常大鼠的痛觉过敏。

关键词: 脑源性神经营养因子, 星形胶质细胞, 痛觉过敏

Abstract:

Objective To investigate the effects of exogenous brain-derived neurotrophic factor (BDNF) on the activation of astrocytes, and analyse the impact of intrathecal injection of activated astrocytes on pain hyperalgia in normal rats. Methods Primary astrocytes cultured in vitro were incubated with exogenous BDNF (100 ng/mL) for 0 min (baseline), 15 min, 60 min and 120 min, and the expression of glial filament acidic protein (GFAP) in cells was detected by Western blotting. Twenty-four male SD rats were randomly divided into activated-astrocytes group (intrathecal injection of astrocytes incubated with BDNF for 15 min), negative control group (intrathecal injection of astrocytes without incubation with BDNF), placebo group (intrathecal injection of PBS) and blank control group after intrathecal catheterization, with 6 rats in each group. Before injection and 0.5 h, 2 h, 4 h and 8 h after injection, values of 50% mechanical paw withdrawal threshold (50% PWT) were measured with von Frey filaments method. Results The expression of GFAP protein in astrocytes after incubation with BDNF for 15 min, 60 min and 120 min was significantly higher than that of the baseline (P<0.01). In activated-astrocytes group, 50%PWT at each time point after injection was significantly lower than that before injection (P<0.01). In negative control group, transient decrease of 50%PWT only occurred 30 min after injection, which was significantly different from that before injection (P<0.01). There was no significant difference between 50%PWT at each time point after injection and that before injection in placebo group and blank control group (P>0.05). Conclusion Exogenous BDNF can activate astrocytes cultured in vitro, and intrathecal injection of exogenous BDNF-activated astrocytes can directly induce pain hyperalgia in normal rats.

Key words: brain-derived neurotrophic factor, astrocyte, pain hyperalgia