Zhao Meng-Meng, Song Qing, Xie Qian-Yun, Sun Wen-Qiang, Zhang Yang, Tang Wei, Li Meng-Xing
College of Acupuncture, Moxibustion and Tuina, Anhui University of Chinese Medicine, Hefei, 230012, China.
Anhui Province Key Laboratory of Meridian Viscera Correlationship, Hefei, 230038, China.
Neurochem Res. 2025 Nov 10;50(6):355. doi: 10.1007/s11064-025-04603-8.
Cerebral ischemia-reperfusion injury (CIRI) represents a critical pathological mechanism underlying ischemic stroke, yet effective therapeutic interventions remain limited. Neurotoxic astrocytes, activated by inflammatory mediators such as interleukin-17 A (IL-17 A), exacerbate neuronal damage. Although electroacupuncture (EA) has demonstrated neuroprotective properties, its influence on IL-17 A signaling and subsequent astrocyte-mediated neurotoxicity in CIRI remains unclear. This study aims to investigate whether EA mitigates CIRI by downregulating IL-17 A to suppress the activation of neurotoxic astrocyte. A mouse model of middle cerebral artery occlusion and reperfusion (MCAO/R) was established employing the Zea-Longa modified ligation method. EA was applied to the Baihui (GV20) and Fengfu (GV16) acupoints. Neurological and behavioral evaluations were performed using the Modified Neurological Severity Score (mNSS), foot fault test, and balance beam test. Cerebral infarction volume was quantified via TTC staining, and neuronal ultrastructure was examined by transmission electron microscopy. Laser speckle imaging was employed to monitor cerebral blood flow before and after modeling and EA treatment. Western blotting was used to analyze protein expression levels of IL-17 A, IL-17RA, NF-κB p65, Bax, Bcl-2, and cleaved-Caspase-3/Caspase-3. Co-localization of IL-17 A with GFAP and C3, as well as IL-17RA with GFAP, was assessed via immunofluorescence staining. qPCR was performed to quantify IL-17 A mRNA levels, while TUNEL staining assessed neuronal apoptosis. ELISA was used to determine the concentrations of IL-17 A, TNF-α, and IL-1β in brain tissue. EA significantly improved neurological function, reduced cerebral infarct size, and alleviated neuronal apoptosis. Compared to the MCAO/R group, EA markedly downregulated IL-17 A expression and its related signaling proteins, inhibited neurotoxic astrocyte activation (C3⁺/GFAP⁺), and suppressed the release of proinflammatory cytokines. Notably, administration of recombinant IL-17 A reversed the neuroprotective effects of EA. These findings suggest that EA mitigates ischemic brain injury by inhibiting IL-17 A-mediated neurotoxic astrocyte activation and neuroinflammation, highlighting its potential as a therapeutic strategy for CIRI.
脑缺血再灌注损伤（CIRI）是缺血性中风的关键病理机制，但有效的治疗干预措施仍然有限。由白细胞介素-17 A（IL-17 A）等炎症介质激活的神经毒性星形胶质细胞会加剧神经元损伤。尽管电针（EA）已显示出神经保护特性，但其对CIRI中IL-17 A信号传导以及随后星形胶质细胞介导的神经毒性的影响仍不清楚。本研究旨在探讨EA是否通过下调IL-17 A来减轻CIRI，以抑制神经毒性星形胶质细胞的激活。采用Zea-Longa改良结扎法建立大脑中动脉闭塞再灌注（MCAO/R）小鼠模型。将EA应用于百会（GV20）和风府（GV16）穴位。使用改良神经功能缺损评分（mNSS）、足错试验和平衡木试验进行神经和行为评估。通过TTC染色定量脑梗死体积，并通过透射电子显微镜检查神经元超微结构。采用激光散斑成像监测建模和EA治疗前后的脑血流量。蛋白质免疫印迹法用于分析IL-17 A、IL-17RA、NF-κB p65、Bax、Bcl-2和裂解型半胱天冬酶-3/半胱天冬酶-3的蛋白表达水平。通过免疫荧光染色评估IL-17 A与GFAP和C3的共定位，以及IL-17RA与GFAP的共定位。进行qPCR以定量IL-17 A mRNA水平，而TUNEL染色评估神经元凋亡。ELISA用于测定脑组织中IL-17 A、TNF-α和IL-1β的浓度。EA显著改善神经功能，减小脑梗死面积，并减轻神经元凋亡。与MCAO/R组相比，EA显著下调IL-17 A表达及其相关信号蛋白，抑制神经毒性星形胶质细胞激活（C3⁺/GFAP⁺），并抑制促炎细胞因子的释放。值得注意的是，给予重组IL-17 A可逆转EA的神经保护作用。这些发现表明，EA通过抑制IL-17 A介导的神经毒性星形胶质细胞激活和神经炎症来减轻缺血性脑损伤，突出了其作为CIRI治疗策略的潜力。
