Fang Hao, Fan Ling-Ling, Ding Ye-Ling, Wu Dan, Zheng Jia-Yi, Cai Ye-Feng, Huang Yan, Qiao Li-Jun, Zhang Shi-Jie, Zhan Jie
State Key Laboratory of Traditional Chinese Medicine Syndrome, The Second Affiliated Hospital of Guangzhou, University of Chinese Medicine, Guangzhou, 510405, China.
Department of Neurology, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510405, China.
Neurochem Res. 2024 Nov;49(11):3105-3117. doi: 10.1007/s11064-024-04228-3. Epub 2024 Aug 21.
Cerebral ischemia reperfusion injury is a severe neurological impairment that occurs after blood flow reconstruction in stroke, and microglia cell pyroptosis is one of its important mechanisms. Electroacupuncture has been shown to be effective in mitigating and alleviating cerebral ischemia reperfusion injury by inhibiting neuroinflammation, reducing cellular pyroptosis, and improving neurological function. In this experiment, we divided the rats into three groups, including the sham operation (Sham) group, the middle cerebral artery occlusion/reperfusion (MCAO/R) group, and the pre-electroacupuncture (EAC) group. Pre-electroacupuncture group was stimulated with electroacupuncture of a certain intensity on the Baihui (GV 20) and Dazhui (GV 14) of the rat once a day from the 7th day to the 1st day before the MCAO/R operation. The extent of cerebral infarction was detected by TTC staining. A modified Zea-Longa five-point scale scoring system was used to determine neurologic function in MCAO rats. The number of neurons and morphological changes were accessed by Nissl staining and HE staining. The cellular damage was detected by TUNEL staining. In addition, the expression levels of RhoA, pyrin, GSDMD, Caspase1, cleaved-Caspase1, Iba-1, CD206, and ROCK2 were examined by western blotting and immunofluorescence. The results found that pre-electroacupuncture significantly attenuated neurological impairment and cerebral infarction compared to the post-MCAO/R rats. In addition, pre-electroacupuncture therapy promoted polarization of microglia to the neuroprotective (M2) phenotype. In addition, pre-electroacupuncture inhibited microglia pyroptosis by inhibiting RhoA/pyrin/GSDMD signaling pathway, thereby reducing neuronal injury and increasing neuronal survival in the MCAO/R rats. Taken together, these results demonstrated that pre-acupuncture could attenuate cerebral ischemia-reperfusion injury by inhibiting microglial pyroptosis. Therefore, pre-electroacupuncture might be a potential preventive strategy for ischemic stroke patients.
脑缺血再灌注损伤是一种严重的神经功能障碍,发生在中风后血流重建时,而小胶质细胞焦亡是其重要机制之一。电针已被证明可通过抑制神经炎症、减少细胞焦亡和改善神经功能来有效减轻和缓解脑缺血再灌注损伤。在本实验中,我们将大鼠分为三组,包括假手术(Sham)组、大脑中动脉闭塞/再灌注(MCAO/R)组和预电针(EAC)组。预电针组在MCAO/R手术前第7天至第1天每天对大鼠的百会(GV 20)和大椎(GV 14)进行一定强度的电针刺激。通过TTC染色检测脑梗死范围。采用改良的Zea-Longa五点量表评分系统来确定MCAO大鼠的神经功能。通过尼氏染色和HE染色评估神经元数量和形态变化。通过TUNEL染色检测细胞损伤。此外,通过蛋白质免疫印迹法和免疫荧光法检测RhoA、吡啉、GSDMD、半胱天冬酶1、裂解的半胱天冬酶1、离子钙结合衔接分子1、CD206和ROCK2的表达水平。结果发现,与MCAO/R术后大鼠相比,预电针显著减轻了神经功能障碍和脑梗死。此外,预电针疗法促进小胶质细胞向神经保护(M2)表型极化。此外,预电针通过抑制RhoA/吡啉/GSDMD信号通路抑制小胶质细胞焦亡,从而减少MCAO/R大鼠的神经元损伤并增加神经元存活。综上所述,这些结果表明预针刺可通过抑制小胶质细胞焦亡减轻脑缺血再灌注损伤。因此,预电针可能是缺血性中风患者的一种潜在预防策略。