Bu Juan, Shi Shen, Wang Hui-Qin, Niu Xiao-Shan, Zhao Zong-Feng, Wu Wei-Dong, Zhang Xiao-Ling, Ma Zhi, Zhang Yan-Jun, Zhang Hui, Zhu Yi
Clinical Research Center, People's Hospital of Xinjiang Uygur Autonomous Region, Urumqi, Xinjiang Uygur Autonomous Region, China.
Laboratory Animal Research Center, Center for Disease Control and Prevention, Xinjiang Uygur Autonomous Region, Urumqi, Xinjiang Uygur Autonomous Region, China.
Neural Regen Res. 2019 Apr;14(4):605-612. doi: 10.4103/1673-5374.247465.
Acacetin (5,7-dihydroxy-4'-methoxyflavone), a potential neuroprotective agent, has an inhibitory effect on lipopolysaccharide-induced neuroinflammatory reactions. However, whether acacetin has an effect on inflammatory corpuscle 3 (NLRP3) after cerebral ischemia-reperfusion injury has not been fully determined. This study used an improved suture method to establish a cerebral ischemia-reperfusion injury model in C57BL/6 mice. After ischemia with middle cerebral artery occlusion for 1 hour, reperfusion with intraperitoneal injection of 25 mg/kg of acacetin (acacetin group) or an equal volume of saline (0.1 mL/10 g, middle cerebral artery occlusion group) was used to investigate the effect of acacetin on cerebral ischemia-reperfusion injury. Infarct volume and neurological function scores were determined by 2,3,5-triphenyltetrazolium chloride staining and the Zea-Longa scoring method. Compared with the middle cerebral artery occlusion group, neurological function scores and cerebral infarction volumes were significantly reduced in the acacetin group. To understand the effect of acacetin on microglia-mediated inflammatory response after cerebral ischemia-reperfusion injury, immunohistochemistry for the microglia marker calcium adapter protein ionized calcium-binding adaptor molecule 1 (Iba1) was examined in the hippocampus of ischemic brain tissue. In addition, tumor necrosis factor-α, interleukin-1β, and interleukin-6 expression in ischemic brain tissue of mice was quantified by enzyme-linked immunosorbent assay. Expression of Iba1, tumor necrosis factor-α, interleukin-1β and interleukin-6 was significantly lower in the acacetin group compared with the middle cerebral artery occlusion group. Western blot assay results showed that expression of Toll-like receptor 4, nuclear factor kappa B, NLRP3, procaspase-1, caspase-1, pro-interleukin-1β, and interleukin-1β were significantly lower in the acacetin group compared with the middle cerebral artery occlusion group. Our findings indicate that acacetin has a protective effect on cerebral ischemia-reperfusion injury, and its mechanism of action is associated with inhibition of microglia-mediated inflammation and the NLRP3 signaling pathway.
刺槐素（5,7 - 二羟基 - 4'- 甲氧基黄酮）是一种潜在的神经保护剂，对脂多糖诱导的神经炎症反应具有抑制作用。然而，刺槐素在脑缺血再灌注损伤后对炎性小体3（NLRP3）是否有影响尚未完全明确。本研究采用改良的缝合方法在C57BL/6小鼠中建立脑缺血再灌注损伤模型。在大脑中动脉闭塞缺血1小时后，通过腹腔注射25 mg/kg刺槐素（刺槐素组）或等体积生理盐水（0.1 mL/10 g，大脑中动脉闭塞组）进行再灌注，以研究刺槐素对脑缺血再灌注损伤的影响。通过2,3,5 - 三苯基四氮唑氯化物染色和Zea - Longa评分法测定梗死体积和神经功能评分。与大脑中动脉闭塞组相比，刺槐素组的神经功能评分和脑梗死体积显著降低。为了解刺槐素对脑缺血再灌注损伤后小胶质细胞介导的炎症反应的影响，对缺血脑组织海马区进行小胶质细胞标志物钙适配蛋白离子化钙结合衔接分子1（Iba1）的免疫组织化学检测。此外，通过酶联免疫吸附测定法定量小鼠缺血脑组织中肿瘤坏死因子 - α、白细胞介素 - 1β和白细胞介素 - 6的表达。与大脑中动脉闭塞组相比，刺槐素组Iba1、肿瘤坏死因子 - α、白细胞介素 - 1β和白细胞介素 - 6的表达显著降低。蛋白质免疫印迹分析结果显示，与大脑中动脉闭塞组相比，刺槐素组Toll样受体4、核因子κB、NLRP3、半胱天冬酶原 - 1、半胱天冬酶 - 1、白细胞介素 - 1β前体和白细胞介素 - 1β的表达显著降低。我们的研究结果表明，刺槐素对脑缺血再灌注损伤具有保护作用，其作用机制与抑制小胶质细胞介导的炎症和NLRP3信号通路有关。
