Zhao Guo-Jian, Zhang Li-Mei, Wang Si-Rou, Yang Mei, Jiang Jia-Hao, Yuan Bo-Xiang, Huang Cheng, Huang Zhi-Hua, Tang Xiao-Lu, Chen Tao
Jiangxi Province Key Laboratory of Pharmacology of Traditional Chinese Medicine, Key Laboratory of Prevention and Treatment of Cardiovascular and Cerebrovascular Diseases of Ministry of Education, Ganzhou Key Laboratory of Neuroinflammation Research, Gannan Medical University, Ganzhou, China.
College of Physical Education and Health Science, Yibin University, Yibin, China.
CNS Neurosci Ther. 2025 Jul;31(7):e70519. doi: 10.1111/cns.70519.
Neuroinflammation constitutes a critical pathological event subsequent to ischemic stroke. AD16, a novel anti-neuroinflammatory compound, has demonstrated efficacy in alleviating neuroinflammation in neonatal rats induced by ischemia-hypoxia. This study aims to elucidate the therapeutic utility and underlying mechanisms of AD16 in an adult ischemic stroke rat model.
A rat transient middle cerebral artery occlusion (tMCAO) model was employed. Neurological function was evaluated using the Longa and Garcia JH scores, motor function was assessed through rotary rod and CatWalk gait analysis, and brain injury was examined via TTC and Nissl staining. Molecular docking techniques simulate the binding of a target compound to a potential target. Western blot, immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) were used to detect microglia phenotype, pro-inflammatory factors, and activation of signaling molecules.
AD16 treatment improved neural function in tMCAO rats, reduced cerebral infarction volume and brain water content, preserved blood-brain barrier integrity, and inhibited pro-inflammatory cytokines. Molecular docking showed AD16 has high affinity for α7nAChR, TLR4, ERK, and STAT3. AD16 increased α7nAChR, CD206, and p-ERK protein levels, while decreasing CD40, CD68, TLR4, and p-STAT3. These effects were reversed by α-BTX (α7nAChR inhibitor) and U0126 (ERK inhibitor).
AD16 may inhibit microglia activation and polarization via the α7nAChR-ERK-STAT3 pathway, thus reducing neuroinflammation from cerebral ischemia and protecting the brain. This study suggests AD16 as a potential treatment for ischemic stroke.
神经炎症是缺血性中风后的一个关键病理事件。AD16是一种新型抗神经炎症化合物，已证明其在减轻新生大鼠缺血缺氧诱导的神经炎症方面具有疗效。本研究旨在阐明AD16在成年缺血性中风大鼠模型中的治疗效用及潜在机制。
采用大鼠短暂性大脑中动脉闭塞（tMCAO）模型。使用Longa和Garcia JH评分评估神经功能，通过转棒试验和CatWalk步态分析评估运动功能，并通过TTC和尼氏染色检查脑损伤。分子对接技术模拟目标化合物与潜在靶点的结合。采用蛋白质印迹法、免疫荧光法和酶联免疫吸附测定（ELISA）检测小胶质细胞表型、促炎因子和信号分子的激活情况。
AD16治疗改善了tMCAO大鼠的神经功能，减少了脑梗死体积和脑含水量，维持了血脑屏障的完整性，并抑制了促炎细胞因子。分子对接显示AD16对α7nAChR、TLR4、ERK和STAT3具有高亲和力。AD16增加了α7nAChR、CD206和p-ERK蛋白水平，同时降低了CD40、CD68、TLR4和p-STAT3。这些作用被α-BTX（α7nAChR抑制剂）和U0126（ERK抑制剂）逆转。
AD16可能通过α7nAChR-ERK-STAT3途径抑制小胶质细胞的激活和极化，从而减轻脑缺血引起的神经炎症并保护大脑。本研究表明AD16有望成为缺血性中风的一种治疗方法。
