哈巴苷和乙酰哈巴苷减轻脑缺血再灌注损伤的机制研究  

Study on the mechanism of harpaside and acetylharpaside for alleviating cerebral ischemia-reperfusion injury

作　　者：邢慧颖 田洋 冷沁洁 刘洋 彭静 李洁 王琦 谭睿[1] XING Huiying;TIAN Yang;LENG Qingjie;LIU Yang;PENG Jing;LI Jie;WANG Qi;TAN Rui(School of Life Sciences and Engineering,Southwest Jiaotong University,Chengdu,Sichuan,610031 P.R.China)

机构地区：[1]西南交通大学生命科学与工程学院,四川成都610031

出　　处：《华西药学杂志》2023年第5期493-497,共5页West China Journal of Pharmaceutical Sciences

基　　金：四川省药品监督管理局民族药标准提升项目(510201202102305);四川省中医药产业发展重大项目(510201202109711);四川省政府应急重大专项(2021XYCZ008)。

摘　　要：目的 研究哈巴苷和乙酰哈巴苷改善大鼠脑缺血再灌注损伤的作用及机制。方法 WB法检测哈巴苷和乙酰哈巴苷对PC12损伤细胞中核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、核因子(NF)-κB、白细胞介素(IL)-1β蛋白含量的变化。将SD大鼠随机分为假手术组、模型组、哈巴苷组(2 mg·kg-1)和乙酰哈巴苷组(2 mg·kg-1),每组6只。采用脑中动脉阻断法造模,评价神经功能评分、脑梗死体积、含水量,HE染色、Nissl染色、TUNEL染色、免疫荧光染色检测大鼠脑组织NLRP3、人半胱氨酸蛋白酶(caspase)-1、IL-1β、NF-κB蛋白表达的水平。结果 哈巴苷和乙酰哈巴苷极显著降低了PC12损伤细胞中NLRP3、IL-1β、NF-κB蛋白的表达量。与模型组比较,两组给药后降低了神经功能评分、脑梗死体积及脑含水量,组织间隙减轻,尼氏体数目增加,神经元凋亡减少,NLRP3、caspese-1、IL-1β、NF-κB蛋白红色荧光减弱。结论 哈巴苷和乙酰哈巴苷可通过抑制NF-κB/NLRP3信号通路,改善脑缺血再灌注损伤大鼠神经元凋亡、减少脑梗死体积、抑制炎症反应、缓解脑缺血再灌注引起的脑损伤。OBJECTIVE To study the effect and mechanism of harpaside and acetylharpaside on cerebral ischemia-reperfusion injury in rats.METHODS The changes of NLRP3,NF-κB and IL-1βprotein contents in PC12 injured cells were detected by Western blot method.SD rats were randomly divided into sham group,model group,har group(2 mg·kg-1)and ace group(2 mg·kg-1),with 6 rats in each group.The model was made by occlusion of middle cerebral artery.The study also utilized evaluation of neurological function score,cerebral infarction volume,water content,HE staining,Nissl staining,TUNEL staining and immunofluorescence staining to detect the expression level of NLRP3/caspase-1/IL-1β/NF-κB proteins in the rat brain tissue.RESULTS The expressions of NLRP3,IL-1βand NF-κB proteins in PC12 injured cells were significantly reduced by harpaside and acetylharpaside.Compared with the model group,the neurological function score,cerebral infarct volume and brain water content were decreased,and the interstitial space was alleviated.The number of Nissl bodies was increased and neuronal apoptosis was reduced.The red fluorescence of NLRP3,caspese-1,IL-1βand NF-κB proteins was weakened.CONCLUSION Harpaside and acetylharpaside can improve neuronal apoptosis in rats with cerebral ischemia-reperfusion injury,reduce cerebral infarct volume,inhibit inflammatory response and alleviate brain injury caused by cerebral ischemia-reperfusion by inhibiting NF-κB/NLRP3 signaling pathway.

关 键 词：哈巴苷 乙酰哈巴苷 PC12损伤细胞 脑缺血再灌注损伤 神经元凋亡 炎症反应 NLRP3/NF-κB信号通路 

分 类 号：R96[医药卫生—药理学]