天津医药 ›› 2017, Vol. 45 ›› Issue (4): 364-367.doi: 10.11958/20170055

• 实验研究 • 上一篇    下一篇

基于不同时间窗的延迟亚低温对颅脑创伤大鼠 Bcl-2、 Bax 和 Caspase-3 表达的影响

赵万勇, 李晓红, 王景景, 徐超, 王丽娜, 陈江龙, 张赛, 孙洪涛△   

  1. 武警后勤学院附属医院, 脑创伤与神经疾病研究所, 天津市神经创伤修复重点实验室 (邮编 300162)
  • 收稿日期:2017-01-12 修回日期:2017-01-24 出版日期:2017-04-15 发布日期:2017-04-15
  • 通讯作者: △通讯作者 E-mail: chenmo333@163.com E-mail:837993342@qq.com
  • 作者简介:赵万勇 (1989), 男, 硕士研究生, 主要从事颅脑创伤相关研究
  • 基金资助:
    国家自然科学基金资助项目(81471275, 81401067, 81671222); 天津市应用基础与前沿技术研究计划(15JCQNJC11100); 天津 市自然科学基金项目 (16JCYBJC27600)

Effects of delayed mild hypothermia based on different time windows on the expressions of Bcl-2, Bax and Caspase-3 after traumatic brain injury in rats

ZHAO Wan-yong, LI Xiao-hong, WANG Jing-jing, XU Chao, WANG Li-na, CHEN Jiang-long, ZHANG Sai, SUN Hong-tao△   

  1. The Affiliated Hospital of Logistics College of PAP, Institute of Traumatic Brain Injury and Neurology, Tianjin Key Laboratory of Neurotrauma Repair, Tianjin 300162, China
  • Received:2017-01-12 Revised:2017-01-24 Published:2017-04-15 Online:2017-04-15
  • Contact: △Corresponding Author E-mail: chenmo333@163.com E-mail:837993342@qq.com

摘要: 目的 探讨不同时间窗的延迟亚低温(MHT)治疗对颅脑创伤(TBI)大鼠脑组织 Bcl-2、 Bax 和 Caspase-3 蛋白表达的影响。方法 36 只清洁级成年雄性 SD 大鼠随机均分为常温治疗 (NT) 组、 MHT 15 min 组、 MHT 2 h 组及 MHT 4 h 组。采用电子可控性皮质损伤装置制备大鼠 TBI 模型, 造模完成后 NT 组给予常温 (37 ℃) 维持 6 h, 3 个亚 低温组分别于 TBI 后 15 min、 2 h、 4 h 给予低温 (33.0±1.0) ℃维持 6 h。3 d 后对各组大鼠进行改良神经功能缺损评分 (mNSS), HE 染色观察海马 CA1 区组织病理学变化, 免疫组化染色和 Western blot 检测 Bcl-2、 Bax 和 Caspase-3 蛋白 表达情况。结果 各组大鼠表现为不同程度的神经行为缺陷, 与 NT 组相比, 3 个亚低温组 mNSS 评分均降低(P < 0.01)。HE 染色结果显示 3 个亚低温组神经细胞结构较规则、 排列相对整齐, 神经元坏死数量减少, 核碎裂、 溶解现 象减轻。与 NT 组相比, 3 个亚低温组均能上调 Bcl-2 表达, 下调 Bax 和 Caspase-3 表达(P < 0.05)。以上实验结果 均显示 MHT 15 min 组疗效优于 MHT 2 h 组, 而 MHT 2 h 组和 MHT4 h 组疗效相当。结论 恰当地延迟亚低温治疗 在一定程度上能够抑制神经细胞凋亡, 缓解脑损伤进展。

关键词: 颅脑损伤, 原癌基因蛋白质 c-bcl-2, bcl-2 相关 X 蛋白质, 半胱氨酸天冬氨酸蛋白酶 3, 亚低温, 时间窗

Abstract: Objective To explore the effects of delayed mild hypothermia (MHT) in different time windows on the expressions of Bcl-2, Bax and Caspase-3 in brain tissue of model rats with traumatic brain injury (TBI). Methods Thirtysix clean adult male SD rats were randomly divided into NT group (normal temperature), MHT 15 min group, MHT 2 h group and MHT 4 h group. TBI rat model was established by electronical controlled cortical injury device. The rats in the NT group were treated with normothermia (37 ℃) and the rats in the three hypothermia groups were implemented with low temperature (33.0±1.0) ℃ at 15 min, 2 h and 4 h for 6 h respectively after establishment of TBI model. The modified neurological senerity scores (mNSS), morphological changes in hippocampal CA1 areas, immunohistochemical staining and Western blot assay for Bcl-2, Bax and Caspase-3 were compared 3 days after TBI between the four groups. Results The neurological behavioral deficits were found in each group. Compared with the NT group, the mNSS were decreased in the three hypothermia groups (P < 0.01). The results of HE staining showed that the structure of neurons was regular and arranged neatly, and the number of neurons decreased with alleviated nuclear fragmentation and dissolution in hypothermia groups. Compared with the NT group, the expression of Bcl- 2 was upregulated, and the expressions of Bax and Caspase- 3 were downregulated in three hypothermia groups (P < 0.05). The above experimental results were superior in MHT15 min group to MHT 2 h group, and the therapeutic effect in MHT 2 h group was similar to MHT 4 h group. Conclusion The proper delayed mild hypothermia treatment could inhibit neuronal apoptosis and alleviate brain damage.

Key words: craniocerebral trauma, proto- oncogene proteins c- bcl- 2, bcl- 2- associated X protein, caspase 3, low temperature, time window