ADS032对急性缺血性脑卒中小鼠的神经保护作用

Neuroprotective effect of ADS032 on acute ischemic stroke in mice

目的探讨ADS032对小鼠急性缺血性脑卒中后神经功能的作用及机制。方法将100只雄性C57BL/6小鼠随机划分为5组:假手术组、模型组及低、中、高剂量ADS032组,每组20只。线栓法构建右侧大脑中动脉栓塞模型模拟缺血性脑卒中,脑缺血后立即腹腔注射ADS032(50、100、200 mg/kg)治疗3 d(1次/d),依据Longa评分评估小鼠神经功能,使用TTC染色评估脑梗死体积,免疫荧光染色法检测梗死侧小胶质细胞含量的变化,利用实时荧光定量PCR检测梗死侧脑组织抗炎因子白细胞介素(IL)-4、IL-10以及促炎因子IL-18、IL-1β的表达,利用免疫印迹法检测NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体、凋亡相关斑点样蛋白(ASC)以及IL-18、IL-1β的表达。结果与假手术组相比,模型组小鼠脑梗死明显(P<0.05),神经功能缺损加重(P<0.05),梗死侧脑组织小胶质细胞数量明显增加(P<0.05),NLRP3、ASC、IL-18、IL-1β水平明显升高(P<0.05),IL-4、IL-10水平明显下降(P<0.05)。与模型组相比,ADS032明显减小脑梗死体积(P<0.05),明显改善神经功能缺损(P<0.05),明显减少梗死侧脑组织小胶质细胞数量(P<0.05),明显减轻神经炎症反应、增加抗炎因子水平(P<0.05),并且呈剂量依赖性。结论ADS032对小鼠急性缺血性脑卒中具有显著的神经保护作用,其机制可能与抑制炎症反应有关。

Objective To explore the effect and mechanism of ADS032 on neurological function in mice after acute ischemic stroke.Methods One hundred male C57BL/6 mice were randomly divided into five groups:sham operation group,model group,and low-,medium-,and high-dose ADS032 groups,with 20 mice in each group.The middle cerebral artery occlusion model was established by suture method to simulate ischemic stroke.After ischemia,ADS032(50,100,200 mg/kg)was intraperitoneally injected once a day for 3 days.The neurological function of mice was evaluated according to the Longa score,the cerebral infarction volume was evaluated by TTC staining,the content of microglia in the infarcted cerebral tissues was detected by immunofluorescence staining,the expressions of anti-inflammatory factors interleukin(IL)-4,IL-10 and pro-inflammatory factors IL-18,IL-1βin the infarcted cerebral tissues were detected by real-time fluorescent quantitative PCR,and the expressions of NOD-like receptor pyrin domain-containing protein 3(NLRP3)inflammasome,apoptosis-associated speck-like protein(ASC),and IL-18,IL-1βwere detected by Western blotting.Results Compared with the sham operation group,the model group had significant cerebral infarction(P<0.05),aggravated neurological deficit(P<0.05),a significant increase in the number of microglia in the infarcted cerebral tissues(P<0.05),significantly increased levels of NLRP3,ASC,IL-18,and IL-1β(P<0.05),and significantly decreased levels of IL-4 and IL-10(P<0.05).Compared with the model group,ADS032 significantly reduced the cerebral infarction volume(P<0.05),significantly improved the neurological deficit(P<0.05),significantly reduced the number of microglia in the infarcted cerebral tissues(P<0.05),significantly alleviated the neuroinflammatory response,and increased the levels of anti-inflammatory factors(P<0.05),in a dose-dependent manner.Conclusion ADS032 has a significant neuroprotective effect on acute ischemic stroke in mice,and its mechanism may be related to the inhibition of inflammatory response.