Hubei Provincial Collaborative Innovation Center of Preventive Treatment by Acupuncture and Moxibustion, Hubei University of Chinese Medicine, Wuhan, 430061, China.
Department of Acupuncture, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
Curr Med Sci. 2020 Dec;40(6):1128-1136. doi: 10.1007/s11596-020-2295-9. Epub 2021 Jan 11.
Acute focal cerebral ischemic stroke (IS) is a leading cause of morbidity and mortality worldwide. Acupuncture is an emerging alternative therapy that has been beneficial to acute brain ischemia. However, the underlying protective mechanism of its neuroprotective effect remains unclear. Human original circadian rhythm will be lost after IS, which seriously affects the quality of life and functional recovery of stroke patients. We hypothesize that acupuncture treats IS by regulating the balance of Clock and Bmal1. This study aims to explore the effect of acupuncture at acupoints GV20 and BL23 on neuroprotection and anti-apoptosis in middle cerebral artery occlusion (MCAO) rats and expression of apoptosis and circadian rhythm related proteins. Male Sprague-Dawley (SD) rats were randomly divided into five groups: normal group (Normal), sham model group (Sham MCAO), MCAO model group (MCAO), sham electroacupuncture group (Sham EA) and electroacupuncture group (EA). The MCAO model was prepared by electrocoagulation. The first acupuncture treatment was performed within 2 h after surgery, and then acupuncture therapy was performed on 1st day, 2nd day and 3rd day respectively. After their neurological examination at 72 h of ischemia, the rats from each group were sacrificed. Triphenyltetrazolium chloride (TTC) staining was used to evaluate the brain infarct size. Ultrastructural observation on cerebral ischemic cortex and serum inflammatory cytokines were evaluated. TUNEL staining was used to detect cell apoptosis of brain tissue. The expression levels of proteins Bax, bcl-2, caspase-3, Clock and Bmal1 in the cerebral ischemic region were detected by immunofluorescence staining. Here, we presented evidence that EA at GV20 and BL23 could significantly improve the neurological deficit score and infarct size, and alleviate the cell apoptosis of brain tissue. Moreover, acupuncture treatment upregulated the anti-apoptotic Bcl-2/Bax ratio and reversed the upregulation of caspase-3 following 72-h cerebral ischemia. In addition, the expression levels of circadian proteins Clock and Bmal1 were upregulated in EA group while compared with MCAO group. Our study demonstrated that acupuncture exerted neuroprotective effect against neuronal apoptosis after stroke and the mechanism might be related with regulation of circadian rhythm proteins Clock and Bmal1.
急性局灶性脑缺血性中风（IS）是全球发病率和死亡率的主要原因。针灸是一种新兴的替代疗法，对急性脑缺血有一定的益处。然而，其神经保护作用的潜在保护机制尚不清楚。IS 后人体原有的昼夜节律会丧失，严重影响中风患者的生活质量和功能恢复。我们假设针灸通过调节 Clock 和 Bmal1 的平衡来治疗 IS。本研究旨在探讨针刺百会穴和脑户穴对大脑中动脉闭塞（MCAO）大鼠神经保护和抗细胞凋亡作用，以及对凋亡和昼夜节律相关蛋白表达的影响。雄性 Sprague-Dawley（SD）大鼠随机分为五组：正常组（Normal）、假手术模型组（Sham MCAO）、MCAO 模型组（MCAO）、假电针组（Sham EA）和电针组（EA）。采用电凝法制备 MCAO 模型。术后 2 h 内进行首次针刺治疗，然后分别在第 1、2、3 天进行针刺治疗。缺血 72 h 后进行神经功能检查，每组大鼠处死。采用氯化三苯基四氮唑（TTC）染色评估脑梗死体积。超微结构观察大脑缺血皮质和血清炎症细胞因子。TUNEL 染色检测脑组织细胞凋亡。免疫荧光染色检测大脑缺血区 Bax、bcl-2、caspase-3、Clock 和 Bmal1 蛋白的表达水平。本研究表明，GV20 和 BL23 的电针治疗可显著改善神经功能缺损评分和梗死体积，减轻脑组织细胞凋亡。此外，针刺治疗可上调抗凋亡 Bcl-2/Bax 比值，并逆转缺血后 72 h caspase-3 的上调。此外，电针组 Clock 和 Bmal1 昼夜节律蛋白的表达水平上调。本研究表明，针刺对中风后神经元凋亡具有神经保护作用，其机制可能与调节昼夜节律蛋白 Clock 和 Bmal1 有关。
