6-姜烯酚通过miRNA-26a-5p/DAPK1减轻大鼠脑缺血再灌注损伤  

6-Shogaol Attenuates Cerebral Ischaemia-reperfusion Injury in Rats via miRNA-26a-5p/DAPK1

作　　者：饶欧阳 李世欣 朱宁 周航向 陶浚泠 李叶红 刘颖 RAO Ou-yang;LI Shi-xin;ZHU Ning;ZHOU Hang-xiang;TAO Jun-ling;LI Ye-hong;LIU Ying(Clinical Medicine School of Guizhou Medical University,Guiyang 550004,China;Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China)

机构地区：[1]贵州医科大学临床医学院,贵阳550004 [2]贵州医科大学附属医院,贵阳550004

出　　处：《科学技术与工程》2024年第27期11594-11603,共10页Science Technology and Engineering

基　　金：贵州省科技厅自然科学计划(黔科合基础-ZK【2023】一般401);贵州医科大学附属医院博士科研启动基金(gyfybsky2021-65)。

摘　　要：为探究6-姜烯酚(6-Shogaol,6-SH)对大鼠脑缺血再灌注损伤(cerebral ischemia-reperfusion injury,CIRI)的保护作用,采用分子对接观察6-SH能否自发性结合microRNA-26a-5p(miRNA-26a-5P),双萤光素报告基因检测验证miRNA-26a-5p与死亡相关蛋白激酶1(ceath associated protein kinase 1,DAPK 1)的靶向关系;线栓法制备大鼠CIRI模型,雄性SD大鼠随机为假手术组、模型组、6-姜烯酚组;于CIRI后72 h进行改良神经功能评分,氯化三苯基四氮唑(triphenyltetrazolium chloride,TTC)染色,评分后采集脑组织标本进行苏木精-伊红(hematoxylin-eosin staining,HE)染色观察神经细胞病理损伤情况,透射电镜观察神经元超微结构及自噬情况,免疫荧光检测自噬标志物,实时荧光定量和蛋白印迹法检测自噬和钙超载相关蛋白和基因。实验结果表明6姜烯酚可上调miRNA-26a-5p的表达,抑制DAPK1的表达,减轻大鼠CIRI后过度自噬和钙超载,发挥神经保护作用。To explore the protective effects of 6-shogaol(6-SH)against cerebral is chemia-reperfusion injury(CIRI)in rats,molecular docking was utilized to investigate the spontaneous binding ability of 6-SH with microRNA-26a-5p(miRNA-26a-5P).Dual luciferase reporter gene assays were employed to validate the targeted relationship between miRNA-26a-5p and death associated protein kinase 1(DAPK1).The CIRI rat model was induced using the suture method,with male SD rats randomly assigned to sham surgery,model,and 6-shogaol groups.Improved neurological function scoring was conducted 72 hours post-CIRI,accompanied by Triphenyltetrazolium chloride(TTC)staining.Post-scoring,brain tissue specimens were collected for hematoxylin-eosin(HE)staining to observe neuronal pathological damage.Transmission electron microscopy was utilized to examine neuronal ultrastructure and autophagy.Immunofluorescence was employed for the detection of autophagic markers,and real-time fluorescence quantification,along with protein immunoblotting,were performed to assess proteins and genes related to autophagy and calcium overload.The results indicate that 6-shogaol can upregulate miRNA-26a-5p expression,inhibit DAPK1 expression,and alleviate excessive autophagy and calcium overload after CIRI in rats,showcasing its neuroprotective properties.

关 键 词：脑缺血再灌注损伤 6-姜烯酚 MIRNA DAPK1 自噬 钙超载 

分 类 号：R743.31[医药卫生—神经病学与精神病学]