中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2016, Vol. 11 ›› Issue (7): 1108-1114.doi: 10.4103/1673-5374.187045

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

脑卒中后缺血半暗区神经元的自噬和凋亡：时间动态变化启示目标靶向神经保护

  

  • 出版日期:2016-07-30 发布日期:2016-07-30
  • 基金资助:
    国家自然科学基金（81460351）；昆明理工大学博士基金（kksy201360112）；云南教育科学研究基金（2014y070）

Dynamic changes in neuronal autophagy and apoptosis in the ischemic penumbra following permanent ischemic stroke

Yi-hao Deng, Hong-yun He, Li-qiang Yang, Peng-yue Zhang*   

  1. Department of Human Anatomy and Histoembryology, Medical School, Kunming University of Science and Technology, Kunming, Yunnan Province, China
  • Online:2016-07-30 Published:2016-07-30
  • Contact: Peng-yue Zhang, Ph.D., zpy19802000@163.com.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, No. 81460351; the Doctoral Foundation of Kunming University of Science and Technology of China, No. KKSY201360112; the Scientific Research Foundation of Yunnan Provincial Department of Education of China, No. 2014Y070.

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摘要：

脑卒中后缺血半暗区神经元自噬和凋亡发生的时间窗尚不明确。为探究半暗区内自噬和凋亡的动态变化，确定脑卒中后临床治疗的靶点，实验通过大脑中动脉栓塞法制备成年SD大鼠永久缺血性脑卒中模型，以LC3-II和cleaved caspase-3为抗体，通过western blot和免疫荧光染色分别检测脑卒中后缺血半暗区细胞自噬和凋亡水平，发现脑卒中后5 h内，缺血半暗区LC3-II和cleaved caspase-3蛋白表达逐渐升高随后下降，以LC3-II蛋白表达降低更显著。脑梗死体积在缺血后1-4 h缓慢扩大，但至5 h后扩张速度明显加快，而神经功能缺损的严重程度与脑梗死体积的扩大正相关。脑脑卒中后5 h，缺血半暗区LC3-II和cleaved caspase-3的表达维持在较高水平，随后两者的表达逐渐降低；脑脑卒中后72 h，LC3-II的表达已降至很低水平，但cleaved caspase-3的表达仍维持在较高水平。说明脑卒中后半暗区内自噬和凋亡呈不同的时间动态变化，启发临床医生应根据各自的激活状态考虑不同的治疗目标靶向。 

orcid: 0000-0002-7981-3092 (Yi-hao Deng)

关键词: 神经再生, 脑损伤, 永久性缺血性脑卒中, 自噬, 凋亡, LC3-II, Caspase-3, 梗死体积, 神经功能缺损评分, 免疫印迹, 免疫荧光法, 动态变化

Abstract: The temporal dynamics of neuronal autophagy and apoptosis in the ischemic penumbra following stroke remains unclear. Therefore, in this study, we investigated the dynamic changes in autophagy and apoptosis in the penumbra to provide insight into potential therapeutic targets for stroke. An adult Sprague-Dawley rat model of permanent ischemic stroke was prepared by middle cerebral artery occlusion. Neuronal autophagy and apoptosis in the penumbra post-ischemia were evaluated by western blot assay and immunofluorescence staining with antibodies against LC3-II and cleaved caspase-3, respectively. Levels of both LC3-II and cleaved caspase-3 in the penumbra gradually increased within 5 hours post-ischemia. Thereafter, levels of both proteins declined, especially LC3-II. The cerebral infarct volume increased slowly 1–4 hours after ischemia, but subsequently increased rapidly until 5 hours after ischemia. The severity of the neurological deficit was positively correlated with infarct volume. LC3-II and cleaved caspase-3 levels were high in the penumbra within 5 hours after ischemia, and after that, levels of these proteins decreased at different rates. LC3-II levels were reduced to a very low level, but cleaved caspase- 3 levels remained high 72 hours after ischemia. These results indicate that there are temporal differences in the activation status of the autophagic and apoptotic pathways. This suggests that therapeutic targeting of these pathways should take into consideration their unique temporal dynamics.

Key words: nerve regeneration, brain injuries, permanent ischemic stroke, autophagy, apoptosis, LC3-II, cleaved caspase-3, infarct volume, neurological deficit score, western blotting, immunofluorescence, dynamic variations