Li Zeyang, Zhao Minghui, Zhang Xiaoqian, Lu Yiran, Yang Yang, Xie Yalong, Zou Zhimiao, Zhou Liang, Shang Runshi, Zhang Limin, Jiang Fengchao, Du Dunfeng, Zhou Ping
Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Key Laboratory of Organ Transplantation, Ministry of Education, NHC Key Laboratory of Organ Transplantation, Key Laboratory of Organ Transplantation, Chinese Academy of Medical Sciences, Wuhan, China.
Front Pharmacol. 2022 Dec 15;13:1080438. doi: 10.3389/fphar.2022.1080438. eCollection 2022.
Cerebral ischemia-reperfusion injury (CIRI) inevitably occurs after vascular recanalization treatment for ischemic stroke. The accompanying inflammatory cascades have a major impact on outcome and regeneration after ischemic stroke. Evidences have demonstrated that TLR/MyD88/NF-κB signaling contributes to CIRI. This study aimed to investigate the druggability of MyD88 in the central nervous system (CNS) and the neuroprotective and anti-neuroinflammatory effects of the MyD88 inhibitor TJ-M2010-5 on CIRI. A middle cerebral artery occlusion (MCAO) model was used to simulate CIRI in mice. BV-2 cells were stimulated with oxygen glucose deprivation/reoxygenation (OGD/R) or lipopolysaccharide, and SH-SY5Y cells were induced by OGD/R . Neurological deficit scores and cerebral infarction volumes were evaluated. Immunofluorescence staining was performed to measure neuronal damage and apoptosis in the brain. The anti-neuroinflammatory effect of TJ-M2010-5 was evaluated by analyzing the expression of inflammatory cytokines, activation of microglia, and infiltration of peripheral myeloid cells. The expression of proteins of the MyD88/NF-κB and ERK pathway was detected by Simple Western. The concentrations of TJ-M2010-5 in the blood and brain were analyzed by liquid chromatography-mass spectrometry. The cerebral infarction volume decreased in mice treated with TJ-M2010-5, with the most prominent decrease being approximately 80% of the original infarction volume. Neuronal loss and apoptosis were reduced following TJ-M2010-5 treatment. TJ-M2010-5 inhibited the infiltration of peripheral myeloid cells and the activation of microglia. TJ-M2010-5 also downregulated the expression of inflammatory cytokines and inhibited the MyD88/NF-κB and ERK pathway. Furthermore, TJ-M2010-5 showed good blood-brain barrier permeability and no neurotoxicity. TJ-M2010-5 has an excellent therapeutic effect on CIRI as a novel CNS drug candidate by inhibiting excessive neuroinflammatory responses.
脑缺血再灌注损伤（CIRI）在缺血性脑卒中血管再通治疗后不可避免地发生。伴随的炎症级联反应对缺血性脑卒中后的结局和再生有重大影响。证据表明，TLR/MyD88/NF-κB信号通路促成了CIRI。本研究旨在探讨MyD88在中枢神经系统（CNS）中的可药用性，以及MyD88抑制剂TJ-M2010-5对CIRI的神经保护和抗神经炎症作用。采用大脑中动脉闭塞（MCAO）模型在小鼠中模拟CIRI。用氧糖剥夺/复氧（OGD/R）或脂多糖刺激BV-2细胞，并用OGD/R诱导SH-SY5Y细胞。评估神经功能缺损评分和脑梗死体积。进行免疫荧光染色以测量脑中的神经元损伤和凋亡。通过分析炎性细胞因子的表达、小胶质细胞的激活和外周髓样细胞的浸润来评估TJ-M2010-5的抗神经炎症作用。通过Simple Western检测MyD88/NF-κB和ERK通路蛋白的表达。通过液相色谱-质谱法分析血液和脑中TJ-M2010-5的浓度。用TJ-M2010-5治疗的小鼠脑梗死体积减小，最显著的减小约为原始梗死体积的80%。TJ-M2010-5治疗后神经元丢失和凋亡减少。TJ-M2010-5抑制外周髓样细胞的浸润和小胶质细胞的激活。TJ-M2010-5还下调炎性细胞因子的表达并抑制MyD88/NF-κB和ERK通路。此外，TJ-M2010-5显示出良好的血脑屏障通透性且无神经毒性。作为一种新型的中枢神经系统候选药物，TJ-M2010-5通过抑制过度的神经炎症反应对CIRI具有优异的治疗效果。
