Dong Shan-Shan, Li Ming-Yue, Yu Xue-Ping, Kan Yu-Na, Dai Xiao-Hong, Zheng Lei, Cao Hong-Tao, Duan Wen-Hui, Luo En-Li, Zou Wei
Acupuncture and Moxibustion Department, First Affiliated Hospital of Heilongjiang University of Chinese Medicine, Harbin, Heilongjiang, P. R. China.
Second Department of Traditional Chinese Medicine, Medical School, South China Hospital, Shenzhen University, Shenzhen, P. R. China.
Brain Behav. 2024 Dec;14(12):e70095. doi: 10.1002/brb3.70095.
Intracerebral hemorrhage (ICH) is a significant global disease with high mortality and disability. As of now, there is no effective therapy available. Oxidative stress and neuronal apoptosis play essential roles in ICH, determining neuronal survival. In our preliminary studies, we found that Baihui-penetrating-Qubin acupuncture could improve neurological deficits and neuropathological damage in the perihematomal area in ICH rats. The SIRT1/FOXO1 signaling pathway has been reported to mediate antioxidant and anti-neuronal apoptosis. This study aimed to investigate the effects of Baihui-penetrating-Qubin acupuncture on oxidative stress and neuronal apoptosis after ICH and the role of SIRT1/FOXO1 in acupuncture's neuroprotection.
ICH rat models were established by autologous tail blood (50 µL) infusion into the caudate nucleus. EX527, SIRT1-specific inhibitor was intraperitoneally administered 3 days before ICH. Baihui-penetrating-Qubin acupuncture treatment was performed once a day for 30 min after ICH. Neurological deficits were evaluated using the modified neurological severity score (mNSS). Brain edema was evaluated using brain water content. HE staining and Nissl staining were used to evaluate neuropathological damage in the perihematomal area. Terminal deoxynucleotidyl transferase dUTP nick end labeling was used to quantify neuronal apoptosis. Specific kits were used to detect the levels of SOD, CAT, GSH-Px in the brain. The oxidative DNA damage was evaluated using enzyme-linked immunosorbent assay to detect the level of 8-hydroxyguanosine (8-OHdG). Western blot was used to evaluate the expressions of SIRT1, Ac-FOXO1, FOXO1, Bcl-2, and Bax. Immunofluorescence staining was conducted to detect the cellular localization of SIRT1.
Baihui-penetrating-Qubin acupuncture improved the neurological deficits and brain edema, reduced the pathological injury and neuronal degeneration in 3 days in the perihematomal area after ICH. Mechanistically, acupuncture reduced oxidative stress injury and neuronal apoptosis via activating SIRT1/FOXO1 pathway. The neuroprotective effects of acupuncture were abolished by injection of the SIRT1 inhibitor EX527.
Baihui-penetrating-Qubin acupuncture could reduce oxidative stress and neuronal apoptosis, at least in part, through the SIRT1/FOXO1 signaling pathway, improving neurological deficits and neuropathological damage after ICH. These findings suggest that Baihui-penetrating-Qubin acupuncture is an effective therapy for ICH, as well as targeting SIRT1 signaling to promote neuron survival could be a potential therapeutic strategy.
脑出血（ICH）是一种全球范围内的重大疾病，具有高死亡率和高致残率。目前尚无有效的治疗方法。氧化应激和神经元凋亡在脑出血中起关键作用，决定着神经元的存活。在我们的初步研究中，我们发现百会透曲鬓针刺可改善脑出血大鼠血肿周围区域的神经功能缺损和神经病理损伤。据报道，SIRT1/FOXO1信号通路介导抗氧化和抗神经元凋亡作用。本研究旨在探讨百会透曲鬓针刺对脑出血后氧化应激和神经元凋亡的影响，以及SIRT1/FOXO1在针刺神经保护中的作用。
采用自体尾血（50 μL）注入尾状核建立脑出血大鼠模型。在脑出血前3天腹腔注射SIRT1特异性抑制剂EX527。脑出血后每天进行一次百会透曲鬓针刺治疗，每次30分钟。采用改良神经功能缺损评分（mNSS）评估神经功能缺损。用脑含水量评估脑水肿。采用苏木精-伊红（HE）染色和尼氏染色评估血肿周围区域的神经病理损伤。采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法（TUNEL）定量神经元凋亡。使用特定试剂盒检测脑中超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GSH-Px）水平。采用酶联免疫吸附测定法检测8-羟基鸟苷（8-OHdG）水平，评估氧化性DNA损伤。采用蛋白质免疫印迹法评估SIRT1、乙酰化FOXO1（Ac-FOXO1）、FOXO1、Bcl-2和Bax的表达。进行免疫荧光染色检测SIRT1的细胞定位。
百会透曲鬓针刺改善了脑出血后3天的神经功能缺损和脑水肿，减轻了血肿周围区域的病理损伤和神经元变性。机制上，针刺通过激活SIRT1/FOXO1通路减轻氧化应激损伤和神经元凋亡。注射SIRT1抑制剂EX527消除了针刺的神经保护作用。
百会透曲鬓针刺可至少部分通过SIRT1/FOXO1信号通路减轻氧化应激和神经元凋亡，改善脑出血后的神经功能缺损和神经病理损伤。这些发现表明，百会透曲鬓针刺是一种治疗脑出血的有效疗法，靶向SIRT1信号通路促进神经元存活可能是一种潜在的治疗策略。
