中国神经再生研究(英文版) ›› 2014, Vol. 9 ›› Issue (15): 1437-1445.doi: 10.4103/1673-5374.139460

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

胡黄连苷Ⅱ保护脑缺血损伤的最佳治疗剂量及时间窗

  

  • 收稿日期:2014-05-06 出版日期:2014-08-12 发布日期:2014-08-12
  • 基金资助:

    国家自然科学基金(81041092, 81274116)

Optimal therapeutic dose and time window of picroside II in cerebral ischemic injury

Guangyi Liu, Li Zhao, Tingting Wang, Meizeng Zhang, Haitao Pei   

  1. Institute of Cerebrovascular Diseases, Affiliated Hospital of Qingdao University, Qingdao, Shandong Province, China
  • Received:2014-05-06 Online:2014-08-12 Published:2014-08-12
  • Contact: Guangyi Liu, Institute of Cerebrovascular Diseases, Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong Province, China, 15898877166@163.com.
  • Supported by:

    This study was financially supported by the National Natural Science Foundation of    China, No. 81041092, 81274116.

摘要:

本课题组前期研究证实,胡黄连苷Ⅱ可抑制脑缺血损伤大鼠缺血半影区神经细胞凋亡,缩小脑梗死体积,改善其神经行为功能。实验旨在通过正交试验法验证胡黄连苷Ⅱ对大鼠脑缺血损伤的神经保护作用,并优化其最佳剂量和时间窗。结果发现,胡黄连苷Ⅱ显示出下调大鼠脑缺血损伤后神经元损伤标志酶神经元特异性烯醇化酶的表达,抑制细胞凋亡作用的最佳治疗时间和剂量经不同检测方法得出:甲苯胺蓝染色显示缺血2.0 h,10 mg/kg;流式细胞术检测显示为缺血1.5 h,10 mg/kg;免疫组织化学染色法显示为缺血2.0 h,10 mg/kg;Western blot检测显示为缺血2.0 h,10 mg/kg;RT-PCR显示为缺血1.5 h,10 mg/kg。实验根据用药剂量最小化和治疗时间窗最大化的原则综合评价显示,胡黄连苷Ⅱ改善和保护脑缺血损伤的最佳治疗时间窗为脑缺血1.5-2.0 h,腹腔注射胡黄连苷Ⅱ剂量为10 mg/kg。

关键词: 神经再生, 胡黄连苷II, 治疗剂量, 时间窗, 脑缺血, NSE, 甲苯胺蓝染色, 流式细胞术, 免疫组织化学, Western blot, RT-PCR, 大鼠, 国家自然科学基金

Abstract:

A preliminary study from our research group showed that picroside II inhibited neuronal apoptosis in ischemic penumbra, reduced ischemic volume, and improved neurobehavioral function in rats with cerebral ischemia. The aim of the present study was to validate the neuroprotective effects of picroside II and optimize its therapeutic time window and dose in a rat model of cerebral ischemia. We found that picroside II inhibited cell apoptosis and reduced the expression of neuron-specific enolase, a marker of neuronal damage, in rats after cerebral ischemic injury. The optimal treatment time after ischemic injury and dose were determined, respectively, as follows: (1) 2.0 hours and 10 mg/kg according to the results of toluidine blue staining; (2) 1.5 hours and 10 mg/kg according to early apoptotic ratio by flow cytometry; (3) 2.0 hours and 10 mg/kg according to immunohistochemical and western blot analysis; and (4) 1.5 hours and 10 mg/kg according to reverse transcription polymerase chain reaction. The present findings suggest that an intraperitoneal injection of 10 mg/kg picroside II 1.5–2.0 hours after cerebral ischemic injury in rats is the optimal dose and time for therapeutic benefit.