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CJAP ›› 2018, Vol. 34 ›› Issue (3): 277-282.doi: 10.12047/j.cjap.5627.2018.065

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Dynamic changes of autophagy activity of cortical neurons in rats with cerebral ischemia and cerebral ischemia/reperfusion

TAO Hong-miao1, SHAN Xiao-yun2, LI Xu-sheng1, CHEN Hao-hao1, MAO Yu-fei1, HE Zhong-ping3   

  1. 1. Jinhua Polytechnic, Jinhua 321000, China;
    2. Jinhua Central Hospital, Jinhua 321000, China;
    3. Jinhua Inspection and Research Institute for Food and Drug Control, Jinhua 321000, China
  • Received:2017-08-28 Revised:2018-02-02 Online:2018-05-28 Published:2018-09-08
  • Supported by:
    浙江省科技厅公益技术应用研究计划项目(2015C37118);金华市科技局社会发展类项目(2015-3-057)

Abstract: Objective: To investigate the changes of autophagy in cerebral cortex neurons at different times after cerebral ischemia and ischemia/reperfusion (I/R) in rats.Methods: Healthy male SD rats (n=60) were randomly assigned into sham-operation group (sham, n=10), cerebral ischemia and ischemia/reperfusion model group (I/R, n=50). In the model group, 10 rats were randomly sampled at five different time points:ischemia 30 min, 2 h, and reperfusion 1 h, 6 h and 24 h after ischemia 2 h. Cerebral infarction volume and cerebral water content were measured. Autophagy of cerebral cortical neurons was examined by transmission electron microscopy (TEM). The protein levels of microtubule associated protein light chain 3-Ⅱ (LC3-Ⅱ) in cerebral cortex of the rats in each group were determined by Western blot.Results: LC3-Ⅱ/Ⅰ ratio had no obvious rise at ischemia 30min,and it began to increase at ischemia 2 h point which was significantly higher than that in sham group (P<0.01).LC3-Ⅱ/Ⅰ ratio was still higher at ischemia/reperfusion 1h and 6h points than that in sham group (P<0.05),although it was decreased comparison with that at ischemia 2 h point. LC3-Ⅱ/Ⅰ ratio reached a peak at ischemia/reperfusion 24 h point, which was significantly higher than that in sham group (P<0.01). This phenomenon was further confirmed by the transmission electron microscopy. The infarction volume at ischemia/reperfusion 6h and 24 h points was increased significantly, and there was a statistical difference compared with sham group (P<0.01). The content of water in the brain tissue of rats at ischemia/reperfusion 24 h was increased obviously, which was significantly higher than that in sham group (P<0.05). Edema, rarefaction, some cell degeneration and apoptosis only in the cortical tissue of rats at ischemia/reperfusion 24 h were showed,and a large number of neurons nuclei in the hippocampus was shrunken,hyperchromatic showing degeneration and apoptosis by HE staining.Conclusion: Neurons autophagy in cerebral cortices of rats with focal cerebral ischemia and ischemia/reperfusion was obviously activated at ischemia 2 h point, it was increased continuously at ischemia/reperfusion 1 h and 6 h points and reached the peak at ischemia/reperfusion 24 h point.

Key words: brain, ischemic/reperfusion injury, LC3, autophagy, cortical neurons, rat

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