法医学杂志

法医学杂志 ›› 2013, Vol. 29 ›› Issue (5): 325-329.DOI: 10.3969/j.issn.1004-5619.2013.05.002

• 论著 • 上一篇    下一篇

成年小鼠氯胺酮慢性中毒后脑细胞凋亡

杨  菊1，李小静1，张志湘2，陆凯明2，卞士中2   

  1. （1. 复旦大学附属上海市第五人民医院病理科，上海 200240； 2. 苏州大学医学部法医系，江苏 苏州 215123）
  • 发布日期:2013-10-25 出版日期:2013-10-28
  • 通讯作者: 卞士中，男，主任法医师，副教授，硕士研究生导师，主要从事法医毒理学研究；E-mail：bianshizhong@suda.edu.cn
  • 作者简介:杨菊（1982—），女，湖北荆州人，硕士，主要从事临床病理学研究；E-mail：yangju_33@126.com

Apoptosis in Adult Mouse Brain after Chronic Poisoning of Ketamine

YANG JU1, LI XIAO-JING1, ZHANG ZHI-XIANG2, LU KAI-MING2, BIAN SHI-ZHONG2   

  1. （1. Department of Pathology, the Fifth People’s Hospital of Shanghai, Fudan University, Shanghai 200240, China; 2. Department of Forensic Medicine, Medical College of Soochow University, Suzhou 215123, China）
  • Online:2013-10-25 Published:2013-10-28

摘要： 目的 研究不同持续时间和剂量下氯胺酮慢性中毒对成年小鼠脑细胞凋亡发生的影响。 方法 氯胺酮按不同剂量（4、10、20、30 mg/kg）每周2次于成年小鼠尾静脉注射，建立小鼠氯胺酮滥用慢性中毒模型，氯胺酮连续注射1、2、4、8、12周后处死。采用透射电镜进行细胞凋亡的定性检测，以Caspase-3免疫荧光染色法和脱氧核糖核苷酸末端转移酶介导的缺口末端标记法（terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling，TUNEL）定量检测凋亡细胞数，推断凋亡发生的时间，并统计分析实验结果。 结果 氯胺酮染毒1周后，在透射电镜下见到脑组织海马及纹状体区域有明显的神经元凋亡，并持续至8周后；染毒1周可见Caspase-3高表达，4周后呈持续低水平表达；染毒1周后可见TUNEL阳性细胞较对照组明显增加，4周时仍处于高水平表达。 结论 氯胺酮尾静脉注射可致成年小鼠神经元凋亡。

关键词: 法医病理学, 中毒, 氯胺酮, 物质滥用, 静脉内, 细胞凋亡, Caspase-3, 小鼠

Abstract: Objective To study the effect of chronic poisoning of ketamine on brain cell apoptosis in adult mouse under different duration and doses. Methods The mouse model of chronic poisoning of ketamine was established on adult mouse by tail vein injection of ketamine twice every week with different doses （4, 10, 20 and 30 mg/kg）. The mice were sacrificed after continuous injection of ketamine of 1, 2, 4, 8 and 12 weeks. The qualitative assessment of apoptosis was made by transmission electron microscope and the quantitative assessment was made by Caspase-3 immumofluorescence staining method and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） to estimate the time point of apoptosis. All the experimental results were statistically analyzed. Results The neuron apoptosis was observed in hippocampus and corpus striatum by transmission electron microscope one week after administration, and continued for eight weeks. High level of Caspase-3 expression was observed one week after administration, but with a low level expression after 4 weeks. The number of TUNEL positive cells obviously increased one week after administration and maintained in a high number at 4 weeks. Conclusion Ketamine by tail vein injection could induce neuron apoptosis in adult mouse.

Key words: forensic pathology, poisoning, ketamine, substance abuse, intravenous, apoptosis, Caspase-3, mice

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