Gu W W, Lu S Q, Ni Y, Liu Z H, Zhou X Y, Zhu Y M, Luo Y, Li X, Li L S, Sun W Z, Zhang H L, Ao G Z
Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases, Department of Pharmacology, Laboratory of Cerebrovascular Pharmacology, College of Pharmaceutical Science, Soochow University, Suzhou 215123, China; Jiangsu Key Laboratory of Preventive and Translational Medicine for Geriatric Diseases, School of Public Health, Soochow University, Suzhou 215123, China.
Department of Emergency, The First Affiliated Hospital of Soochow University, Suzhou 215006, China.
Neuroscience. 2016 Mar 1;316:26-40. doi: 10.1016/j.neuroscience.2015.11.052. Epub 2015 Nov 30.
2-(3',5'-Dimethoxybenzylidene) cyclopentanone (DMBC) is a novel small-molecule compound synthesized by our group. Here, we found that in rat models of permanent middle cerebral artery occlusion (pMCAO), intraperitoneal injection (ip) of DMBC at 1h after ischemia reduced infarct volume, improved neurological deficits and increased the protein levels of microtubule-associated protein 2 (MAP 2) and glial fibrillary acid protein (GFAP) in the ischemic cortex. Post-treatment of DMBC still produced neuroprotective effects even when administered at 6h after ischemia. In the oxygen-glucose deprivation (OGD)-induced astrocytes or HT22 cell injury, DMBC treatment decreased the OGD-induced lactate dehydrogenase (LDH) leakage and increased the GFAP levels in astrocytes. In addition, Annexin-V-Fluos staining analysis revealed that DMBC treatment attenuated both OGD-induced apoptosis and necrosis in astrocytes. Western blotting analysis showed DMBC treatment inhibited the ischemia or OGD-induced increases in active cathepsin B in the ischemic cortex or in astrocytes or HT22 cells. Immunofluorescence analysis demonstrated that DMBC treatment blocked the ischemia or OGD-induced release of cathepsin B from the lysosomes into the cytoplasm in the ischemic cortex or in astrocytes or HT22 cells. Taken together, our results indicate that DMBC can offer neuroprotective effects against cerebral ischemia with an extended therapeutic window and its mechanism might be associated with inhibition of the cathepsin B activation.
2-(3',5'-二甲氧基亚苄基)环戊酮（DMBC）是我们团队合成的一种新型小分子化合物。在此，我们发现，在永久性大脑中动脉闭塞（pMCAO）大鼠模型中，缺血后1小时腹腔注射（ip）DMBC可减小梗死体积，改善神经功能缺损，并增加缺血皮层中微管相关蛋白2（MAP 2）和胶质纤维酸性蛋白（GFAP）的蛋白水平。即使在缺血后6小时给药，DMBC的治疗仍产生神经保护作用。在氧-葡萄糖剥夺（OGD）诱导的星形胶质细胞或HT22细胞损伤中，DMBC处理可减少OGD诱导的乳酸脱氢酶（LDH）泄漏，并增加星形胶质细胞中的GFAP水平。此外，膜联蛋白V-荧光染色分析显示，DMBC处理可减轻OGD诱导的星形胶质细胞凋亡和坏死。蛋白质印迹分析表明，DMBC处理可抑制缺血或OGD诱导的缺血皮层、星形胶质细胞或HT22细胞中组织蛋白酶B活性的增加。免疫荧光分析表明，DMBC处理可阻断缺血或OGD诱导的组织蛋白酶B从溶酶体释放到缺血皮层、星形胶质细胞或HT22细胞的细胞质中。综上所述，我们的结果表明，DMBC可在较宽的治疗窗内对脑缺血提供神经保护作用，其机制可能与抑制组织蛋白酶B的激活有关。
