大鼠继发性脊髓损伤中半胱氨酸蛋白酶水平的变化

doi:吉林省科技厅

大鼠继发性脊髓损伤中半胱氨酸蛋白酶水平的变化

寇万福¹, 杨有赓^1*, 侯元², 刘永茂², 白云深¹

1. 吉林大学第二医院骨科，吉林长春130041；2. 吉林大学基础医学院病原免疫细胞遗传学实验中心, 吉林长春130021

收稿日期:2004-08-25 修回日期:1900-01-01 出版日期:2005-07-28 发布日期:2005-07-28
通讯作者: 杨有赓

Changes of Calpain levels in secondary spinal cord injury in rats

KOU Wan-fu¹, YANG You-geng^1*, HOU Yuan², LIU Yong-mao²,BAI Yun-shen¹

1. Department of Orthopedics, Second Hospital ,Jilin University,Changchun 130041, China;2. Experimental Center of Pathogenobiology, Immunology, Cell Biology and Genitics,School of Basic Medical Sciences,Jilin University,Changchun 130021,China

Received:2004-08-25 Revised:1900-01-01 Online:2005-07-28 Published:2005-07-28
Contact: YANG You-geng

摘要/Abstract

摘要： 目的：通过半胱氨酸蛋白酶（Calpain）的活性变化探讨其在急性脊髓损伤中的作用。方法：将18只雌性SD大鼠随机分为Calpain高选择性抑制剂组(EST)、二甲基亚砜赋形剂对照组（DMSO）和空白对照组（Sham)，每组6只。3组大鼠通过Allen′s方法（50 g-cm)制作急性脊髓损伤模型后，在15 min内，EST组给予EST（1 mg·kg^-1)，DMSO组给予1.5% DMSO，Sham组仅行椎板切除。24 h后每组收集脊髓标本（长度约5.0 cm），分成均等的5份：S₁远头侧、S₂近头侧、S₃损伤段、S₄近尾侧和S₅远尾侧。通过Western blotting对5份样品中的相对分子质量为 68 000神经丝蛋白（NFP）进行分析。结果：S₁~S₅段EST组和Sham组NFP条带密度值显著高于DMSO组（P<0.005），EST组与Sham组比较差异无显著性（P>0.05）；Western blotting条带分析S₃段EST组NFP条带密度值较DMSO组高9.55，S₄段高11.66（P<0.01）。结论：Calpain在急性脊髓损伤中活性显著增高，加重脊髓损伤。

关键词: 长配因, 脊髓损伤, 高选择性抑制剂, 神经丝蛋白

Abstract: Changes of Calpain levels in secondary spinal cord injury in rats KOU Wan-fu¹, YANG You-geng^1*, HOU Yuan², L IU Yong-mao²,BAI Yun-shen¹ ( 1. Department of Orthopedics, Second Objective To observe the changes of activity of calpain in acute spinal cord injury with its highly selective inhibitor EST. Methods Methods:Eighteen S-D female rats were used to set up acute spinal cord injury models with Allen′s method. In 15 min, EST (1 mg·kg^-1) was administrated with intravenous injection in EST group,1.5% Climethyl sulfoxide (DMSO) was given in DMSO group, the rats in sham control group received nothing with laminectomy only. After 24 h, all rats were sacrificed and per fused via the left ventricle with 100 mL of phosphate buffered saline (PBS). Then a 5.0 cm long spinal cord section with the lesion in the center was collected from rat under anesthesia, and divided equally into 1.0 cm segments, S₁ distant rostrol,S₂ near rostrol,S₃ lesion,S₄ near caudal,S₅ distant caudal, which were prepared for Western blotting analysis. Results The density values of neurofilament protein (NFP,68 000) band in EST group and sham group were significantly higher than that in DMSO group (P<0.005). EST could inhibit the degradation of NFP in various phases, the content of NFP increased significantly. Compared with DMSO group, the content of NFP in S₃ section increased 9.55, while that in S₄ section increased 11.66%(P<0.001). Conclusion Calpain activity is significantly enhanced in the rats following acute spinal cord injury. Key words：cysteine endopeptidases; calpain; spinal cord injur ies; highly selective inhibitor；neurofilament protein

Key words: calpain, spinal cord injuries, highly selective inhibitor, neurofilament protein

中图分类号:

R-332， Q556.9

寇万福, 杨有赓, 侯元, 刘永茂, 白云深. 大鼠继发性脊髓损伤中半胱氨酸蛋白酶水平的变化[J]. J4, 2005, 31(4): 574-576.

KOU Wan-fu, YANG You-geng, HOU Yuan, LIU Yong-mao,BAI Yun-shen. Changes of Calpain levels in secondary spinal cord injury in rats[J]. J4, 2005, 31(4): 574-576.

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