Zhang Gao-Feng, Yang Pei, Yin Zeng, Chen Huai-Long, Ma Fu-Guo, Wang Bin, Sun Li-Xin, Bi Yan-Lin, Shi Fei, Wang Ming-Shan
Department of Anesthesiology, Affiliated Qingdao Municipal Hospital of Qingdao University, Qingdao, Shandong Province, China.
Department of Public Health, Affiliated Qingdao Municipal Hospital of Qingdao University, Qingdao, Shandong Province, China.
Neural Regen Res. 2018 Jan;13(1):86-93. doi: 10.4103/1673-5374.224373.
Electroacupuncture preconditioning at acupoint Baihui (GV20) can reduce focal cerebral ischemia/reperfusion injury. However, the precise protective mechanism remains unknown. Mitochondrial fission mediated by dynamin-related protein 1 (Drp1) can trigger neuronal apoptosis following cerebral ischemia/reperfusion injury. Herein, we examined the hypothesis that electroacupuncture pretreatment can regulate Drp1, and thus inhibit mitochondrial fission to provide cerebral protection. Rat models of focal cerebral ischemia/reperfusion injury were established by middle cerebral artery occlusion at 24 hours after 5 consecutive days of preconditioning with electroacupuncture at GV20 (depth 2 mm, intensity 1 mA, frequency 2/15 Hz, for 30 minutes, once a day). Neurological function was assessed using the Longa neurological deficit score. Pathological changes in the ischemic penumbra on the injury side were assessed by hematoxylin-eosin staining. Cellular apoptosis in the ischemic penumbra on the injury side was assessed by terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling staining. Mitochondrial ultrastructure in the ischemic penumbra on the injury side was assessed by transmission electron microscopy. Drp1 and cytochrome c expression in the ischemic penumbra on the injury side were assessed by western blot assay. Results showed that electroacupuncture preconditioning decreased expression of total and mitochondrial Drp1, decreased expression of total and cytosolic cytochrome c, maintained mitochondrial morphology and reduced the proportion of apoptotic cells in the ischemic penumbra on the injury side, with associated improvements in neurological function. These data suggest that electroacupuncture preconditioning-induced neuronal protection involves inhibition of the expression and translocation of Drp1.
百会穴（GV20）电针预处理可减轻局灶性脑缺血/再灌注损伤。然而，确切的保护机制尚不清楚。动力相关蛋白1（Drp1）介导的线粒体分裂可引发脑缺血/再灌注损伤后的神经元凋亡。在此，我们检验了以下假设：电针预处理可调节Drp1，从而抑制线粒体分裂以提供脑保护。通过大脑中动脉闭塞建立局灶性脑缺血/再灌注损伤大鼠模型，在连续5天对GV20进行电针预处理（深度2mm，强度1mA，频率2/15Hz，30分钟，每天1次）后24小时进行。使用Longa神经功能缺损评分评估神经功能。通过苏木精-伊红染色评估损伤侧缺血半暗带的病理变化。通过末端脱氧核苷酸转移酶介导的dUTP-地高辛标记染色评估损伤侧缺血半暗带的细胞凋亡。通过透射电子显微镜评估损伤侧缺血半暗带的线粒体超微结构。通过蛋白质免疫印迹法评估损伤侧缺血半暗带中Drp1和细胞色素c的表达。结果显示，电针预处理降低了总Drp1和线粒体Drp1的表达，降低了总细胞色素c和胞质细胞色素c的表达，维持了线粒体形态，减少了损伤侧缺血半暗带中凋亡细胞的比例，并改善了神经功能。这些数据表明，电针预处理诱导的神经元保护涉及抑制Drp1的表达和转位。
