迷迭香酸通过抑制ROS/NLRP-3炎症小体信号通路对脑缺血再灌注小鼠的保护作用  

Rosmarinic acid protected against brain injury in mice with cerebral ischemia reperfusion via oxidative stress and ROS/NLRP-3 signal pathway inhibition

作　　者：刘勇[1] 杨涛[1] 梁艳山 曹兴华[1] 柯雪茹[1] 陈杰 马晓媛 LIU Yong;YANG Tao;LIANG Yanshan;CAO Xinghua;KE Xueru;CHEN Jie;MA Xiaoyuan(Department of Surgical Anesthesiology,The Affiliated Hospital of Traditional Chinese Medicine,Xinjiang Medical University,Urumqi,Xinjiang 830099,China)

机构地区：[1]新疆医科大学附属中医医院手术麻醉科,乌鲁木齐830099

出　　处：《重庆医学》2023年第19期2905-2911,共7页Chongqing medicine

基　　金：新疆维吾尔自治区自然科学基金项目(2022D01C546)。

摘　　要：目的探讨不同剂量迷迭香酸对脑缺血再灌注小鼠活性氧自由基(ROS)/Nod样受体蛋白-3(NLRP-3)炎症小体信号通路表达的影响,以及迷迭香酸抑制脑缺血再灌注损伤的作用机制。方法60只雄性美国癌症研究所(ICR)小鼠分为假手术组、模型组、迷迭香酸10、20、40 mg/kg组,每组12只。采用改良的Longa线栓法,缺血60 min后进行再灌注建立急性局灶性脑缺血小鼠模型,再灌注后即刻腹腔注射迷迭香酸10、20、40 mg/kg,小鼠清醒后行神经功能学评分,24 h后取脑组织标本,通过苏木素-伊红(HE)染色法观察小鼠脑组织形态学变化,通过免疫荧光分析测定受损侧脑组织标本中ROS水平,Western blot检测超氧化物歧化酶(SOD)1、SOD2、血红素氧合酶-1(HO-1)、核因子-E2相关因子-2(Nrf-2)、NLRP-3、凋亡相关斑点样蛋白(ASC)、caspase-1、白细胞介素-1β(IL-1β)相对表达水平。结果假手术组术后神经功能学评分均为0分。与模型组比较,迷迭香酸10、20、40 mg/kg组神经功能学评分降低,其中迷迭香酸40 mg/kg组降低最明显(P<0.05)。与假手术组比较,模型组脑损伤病理评分、ROS水平增高(P<0.05);与模型组比较,迷迭香酸20 mg/kg组ROS水平降低,迷迭香酸40 mg/kg组脑损伤病理评分、ROS水平降低,其中迷迭香酸40 mg/kg组降低最明显(P<0.05)。与假手术组比较,模型组SOD1、SOD2、HO-1、Nrf-2相对表达水平降低(P<0.05);与模型组比较,迷迭香酸20 mg/kg组SOD1、HO-1、Nrf-2相对表达水平升高,迷迭香酸40 mg/kg组SOD1、SOD2、HO-1、Nrf-2相对表达水平升高,其中迷迭香酸40 mg/kg组升高最明显(P<0.05)。与假手术组比较,模型组NLRP-3、ASC、caspase-1、IL-1β相对表达水平升高(P<0.05);与模型组比较,迷迭香酸10、20、40 mg/kg组NLRP-3、ASC、caspase-1、IL-1β相对表达水平降低,其中迷迭香酸40 mg/kg组降低最明显(P<0.05)。结论迷迭香酸可下调ROS/NLRP-3炎症小体信号通路,抑制炎性及氧化应激Objective To observe the effect of different doses of rosmarinic acid on the expression of inflammatory signal pathway of reactive oxygen species(ROS)/Nod like receptor protein-3(NLRP-3)in mice with cerebral ischemia-reperfusion injury,and to explore the mechanism of rosmarinic acid in inhibiting cerebral ischemia-reperfusion injury.Methods Sixty male American Institute for Cancer Research(ICR)mice were divided into the sham group,the model group,the rosmarinic acid 10,20,40 mg/kg group,with 12 mice in each group.A mouse model of acute focal cerebral ischemia was established by reperfusion after 60 minutes of ischemia with modified Longa suture method.Rosmarinic acid 10,20,40 mg/kg was injected intraperitoneally immediately after reperfusion.Neurofunctional scores were performed after the mice waking,and brain tissue samples were collected 24 h later.The morphological changes of mouse brain tissue were observed by hematoxylin-eosin(HE)staining,and the ROS levels in the damaged side brain tissue samples were determined by immunofluorescence analysis.Western blot analysis of superoxide dismutase(SOD)1,SOD2,heme oxygenase-1(HO-1),nuclear factor E2 associated factor 2(Nrf-2),NLRP-3,apoptosis-associated motle-like protein(ASC),cysteinyl aspartate specific proteinase(caspase)-1,interleukin-1β(IL-1β)relative expression level.Results The neurofunctional scores of the sham group were 0.Compared with the model group,the neurofunctional scores of the rosmarinic acid 10,20,40 mg/kg group were decreased,and the rosmarinic acid 40 mg/kg group had the most significant decrease(P<0.05).Compared with sham operation group,brain injury pathological score and ROS levels were increased in model group(P<0.05).Compared with the model group,ROS level in the rosmarinic acid 20 mg/kg group was decreased,brain injury pathological score and ROS level in the rosmarinic acid 40 mg/kg group were decreased,and the rosmarinic acid 40 mg/kg group had the most significant decrease(P<0.05).Compared with the sham group,the relative expression leve

关 键 词：迷迭香酸 脑缺血再灌注损伤 ROS/NLRP-3信号通路 炎症小体 活性氧 自由基 脑保护 

分 类 号：R285.5[医药卫生—中药学]