Department of Anesthesiology, The First Affiliated Hospital of Wenzhou Medical University, Nanbaixiang, Wenzhou City, Zhejiang Province, 325000, PR China; Department of Anesthesiology, The People's Hospital of Wencheng, Wencheng County, Wenzhou City, Zhejiang Province, 325300, PR China.
Department of Anesthesiology, The First Affiliated Hospital of Wenzhou Medical University, Nanbaixiang, Wenzhou City, Zhejiang Province, 325000, PR China.
Brain Res Bull. 2020 May;158:90-98. doi: 10.1016/j.brainresbull.2020.03.002. Epub 2020 Mar 3.
Electroacupuncture (EA), a traditional Chinese replacement therapy, is widely accepted to treat ischemic stroke. Increasing evidence show that autophagy is involved in the process of cerebral ischemia injury and the Wnt/GSK3β pathway, playing an important role in protecting central nervous system. In this study, rats were treated with EA prior to focal ischemia by middle cerebral artery occlusion (MCAO). Deficit score, infarct volumes and levels of autophagy markers, such as LC3I, LC3II and p62, were assessed with either PI3K inhibitor wortmannin or a GSK-3β inhibitor LiCl. Oxygen-glucose deprivation/re-oxygenation (OGD/R) was made in the primitive neuron in vitro, and was respectively treated with autophagy inhibitors 3-MA, LiCl, GSK3β siRNA, or mTOR inhibitor rapamycin. The results indicated that EA pretreatment increased the levels of autophagy marker LC3-II and reduced the levels of p62. Meanwhile, deficit outcome was improved, and infarct volumes were reduced by EA pretreatment. Furthermore, the beneficial effects of EA pretreatment were reversed by wortmannin. LiCl and GSK3β siRNA can mimic the neuroprotective effects of EA pretreatment by downregulating autophagy, and increasing protein levels of p-mTOR, p-GSK3β and β-catenin in OGD/R neurons. However, the protective effects of GSK3β siRNA were blocked by rapamycin. These results suggest that EA pretreatment induces tolerance to cerebral ischemia by inhibiting autophagy via the Wnt pathway through the inhibition of GSK3β.
电针（EA）是一种传统的中医替代疗法，被广泛用于治疗缺血性中风。越来越多的证据表明，自噬参与脑缺血损伤过程和 Wnt/GSK3β 通路，在保护中枢神经系统方面发挥着重要作用。在这项研究中，通过大脑中动脉闭塞（MCAO）对大鼠进行电针治疗前的局灶性缺血。通过 PI3K 抑制剂wortmannin 或 GSK-3β 抑制剂 LiCl 评估缺血症评分、梗死体积和自噬标志物 LC3I、LC3II 和 p62 的水平。在体外原代神经元中进行氧葡萄糖剥夺/再复氧（OGD/R），并分别用自噬抑制剂 3-MA、LiCl、GSK3β siRNA 或 mTOR 抑制剂 rapamycin 进行处理。结果表明，电针预处理增加了自噬标志物 LC3-II 的水平，降低了 p62 的水平。同时，电针预处理改善了缺血症结果，减少了梗死体积。此外，电针预处理的有益作用可被 wortmannin 逆转。LiCl 和 GSK3β siRNA 可以通过下调自噬并增加 OGD/R 神经元中 p-mTOR、p-GSK3β 和 β-连环蛋白的蛋白水平，模拟电针预处理的神经保护作用。然而，GSK3β siRNA 的保护作用被 rapamycin 阻断。这些结果表明，电针预处理通过抑制 GSK3β 抑制自噬，通过 Wnt 通路诱导脑缺血耐受。
