异甘草素对脑缺血再灌注损伤大鼠Notch信号通路的影响  

Effects of isoliquiritigenin on Notch signaling pathway in rats with cerebral ischemia-reperfusion injury

作　　者：刘成 常丽莉 王强[3] 陈丽薇[1] 祁亚伟 张世军[5] LIU Cheng;CHANG Lili;WANG Qiang;CHEN Liwei;QI Yawei;ZHANG Shijun(Department of Neurology,Yellow River Sanmenxia Hospital,Sanmenxia,Henan 472000,China;Department of Blood Transfusion,Yellow River Sanmenxia Hospital,Sanmenxia,Henan 472000,China;Cardiac Intensive Care Unit,Yellow River Sanmenxia Hospital,Sanmenxia,Henan 472000,China;Department of Neurology,People's Hospital Affiliated to Henan Academy of Medical Sciences,Zhengzhou,Henan 450000,China;Department of Neurosurgery,Yellow River Sanmenxia Hospital,Sanmenxia,Henan 472000,China)

机构地区：[1]黄河三门峡医院神经内科,河南三门峡472000 [2]黄河三门峡医院输血科,河南三门峡472000 [3]黄河三门峡医院心脏重症监护室,河南三门峡472000 [4]河南省医学科学院附属人民医院神经内科,河南郑州450000 [5]黄河三门峡医院神经外科,河南三门峡472000

出　　处：《中华神经外科疾病研究杂志》2025年第2期37-44,共8页Chinese Journal of Neurosurgical Disease Research

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20230950)。

摘　　要：目的探究异甘草素对脑缺血再灌输损伤的保护作用及其分子生物学机制。方法用不同剂量异甘草素(0、10、20、40、80μmol/L)干预小鼠神经干细胞C17.2,进行神经干细胞成球实验,比较细胞成球数;144只大鼠建立脑缺血再灌注大鼠模型,随机分为6组,每组24只,分别为:对照组,脑缺血再灌注损伤组,低、中、高剂量异甘草素治疗组和阳性对照组(尼莫地平),药物干预14 d,检测大鼠脑梗死体积及脑含水量,HE染色检测脑部病理损伤,采用神经功能评分(NDS)系统检测大鼠神经功能;流式细胞术和TUNEL法染色分别检测细胞凋亡;ELISA法检测脑组织中白介素(IL)1β、肿瘤坏死因子α(TNF-α)和IL-10表达水平,化学比色法测定脑组织中超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)含量;免疫印迹法检测Notch1、Notch受体细胞内结合域1(NICD1)和Hes1蛋白相对表达量;另设高剂量异甘草素治疗+DAPT(Notch信号通路抑制剂)组6只,采用上述相同方法检测。结果异甘草素剂量分别为40、80μmol·L-1时,C17.2细胞成球数显著增加(P<0.05);经中、高剂量异甘草素干预后,脑缺血再灌注大鼠脑梗死体积减少,神经损伤评分(NDS)评分降低,脑含水量均降低,大鼠脑组织细胞凋亡率降低,IL-1β、TNF-α、MDA水平降低,IL-10、SOD、GSH水平升高,中、高剂量异甘草素组Notch1、NICD1、Hes1蛋白相对表达量与脑缺血再灌注组大鼠比较均上调,且高剂量组优于中剂量组,而加入DAPT能够逆转高剂量异甘草素的上述作用。结论异甘草素对脑缺血再灌注损伤具有保护作用,其作用机制可能与调控Notch信号通路相关。Objective To explore the protective effects of isoliquiritigenin on cerebral ischemia-reperfusion injury and its molecular biological mechanism.Methods Different doses of isoliquiritigenin(0,10,20,40 and 80μmol/L)were used to intervene rat neural stem cell C17.2,and the neural stem cell sphere-forming assay was carried out to compare the number of cell spheres.144 rats were established as rat models of cerebral ischemia-reperfusion and were randomly divided into 6 groups with 24 rats in each group,namely the control group,the cerebral ischemia-reperfusion injury group,the low-dose isoliquiritigenin treatment group,the middle-dose isoliquiritigenin treatment group,the high-dose isoliquiritigenin treatment group and positive control group(nimodipine).After 14 d of drug intervention,the cerebral infarction volume and cerebral water content in rats were detected.HE staining was used for cerebral pathological injury and neurodeficit score(NDS)system for neurological function detection.Flow cytometry and TUNEL staining were used to assess the cell apoptosis.ELISA was applied to detect the expression levels of interleukin 1β(IL-1β),tumor necrosis factorα(TNF-α)and IL-10 in cerebral tissues and chemical colorimetry was used to measure the contents of superoxide dismutase(SOD),malondialdehyde(MDA)and glutathione(GSH)in cerebral tissues.Western blot was used to detect the protein relative expressions of Notch1,Notch intracellular domain 1(NICD1)and Hes1.6 rats in the high-dose isoliquiritigenin treatment group were further given DAPT(Notch signaling pathway inhibitor)and detected by the same method as above.Results When the dose of isoliquiritigenin was 40μmol·L-1 or 80μmol·L-1,the number of C17.2 cell spheres increased significantly(P<0.05).After intervention of middle and high doses of isoliquiritigenin,the volume of cerebral infarction in rats with cerebral ischemia-reperfusion was reduced,the NDS score was decreased,cerebral water content was lowered,the apoptosis rate of cerebral tissue was declined,the level

关 键 词：异甘草素 脑缺血再灌注 NOTCH信号通路 神经细胞凋亡 缺血后炎症 

分 类 号：R743.3[医药卫生—神经病学与精神病学]