人参皂苷Rb1调控小胶质细胞极化减轻小鼠脑缺血损伤  

Ginsenoside Rb1 alleviates cerebral ischemic injury in mice by regulating microglial polarization

作　　者：刘若静 赵雪 朱毅祯 付玲玲 朱俊德 Liu Ruojing;Zhao Xue;Zhu Yizhen;Fu Lingling;Zhu Junde(Human Anatomy Teaching and Research Laboratory,School of Basic Medical Science,Guizhou Medical University,Gui’an 561113,Guizhou Province,China;Key Laboratory of Brain Function and Diseases Tissue Bank of Higher Education Institutions in Guizhou Province,Gui’an 561113,Guizhou Province,China;Guiqian International General Hospital,Guiyang 550024,Guizhou Province,China;Class 5,School of Clinical Medicine,Guizhou Medical University,Gui’an 561113,Guizhou Province,China)

机构地区：[1]贵州医科大学基础医学院人体解剖学教研室,贵州省贵安新区561113 [2]贵州省高等学校功能与疾病人脑组织库重点实验室,贵州省贵安新区561113 [3]贵黔国际总医院,贵州省贵阳市550024 [4]贵州医科大学,贵州省贵安新区561113

出　　处：《中国组织工程研究》2025年第29期6219-6227,共9页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81660243),项目负责人:朱俊德;贵州省自然科学基金项目(黔科合基础-ZK[2023]一般323),项目负责人:朱俊德;贵州医科大学国家自然科学基金培育项目(20NSP006),项目负责人:朱俊德。

摘　　要：背景:课题组前期研究证明,人参皂苷Rb1改善脑缺血再灌注损伤的神经保护作用可能与Wnt/β-catenin信号通路相关,但具体作用机制仍不清楚。目的:探讨人参皂苷Rb1减轻小鼠脑缺血再灌注损伤的分子机制。方法:将100只C57BL/6小鼠随机分为4组:假手术组(n=25)不建模,脑缺血再灌注损伤组(n=25)采用线栓法制备小鼠大脑中动脉阻塞模型,人参皂苷Rb1组(n=25)采用线栓法制备小鼠大脑中动脉阻塞模型前3 d每天腹腔注射人参皂苷Rb1,人参皂苷Rb1+抑制剂组(n=25)采用线栓法制备小鼠大脑中动脉阻塞模型前3 d每天腹腔注射人参皂苷Rb1与Wnt/β-catenin信号通路抑制剂XAV939。造模3 d后,采用Zea Longa评分、平衡木测试评估小鼠神经功能缺损,TTC染色观察脑梗死体积,干湿质量法检测小鼠脑水肿程度,比色法检测缺血侧脑顶叶超氧化物歧化酶、谷胱甘肽过氧化物酶活性及丙二醛浓度,免疫荧光检测小胶质细胞标志物Iba1与诱导型一氧化氮合成酶(或精氨酸酶1)共表达情况,Western Blot检测水通道蛋白AQP4、炎症相关因子及Wnt/β-catenin通路蛋白糖原合酶激酶3β、β-catenin磷酸化水平,q-PCR检测炎症因子mRNA表达。结果与结论:①与脑缺血再灌注损伤组相比,人参皂苷Rb1组小鼠神经功能缺损症状、脑梗死灶及脑组织水肿、氧化应激与炎症反应均减轻,M1型(Iba1与诱导型一氧化氮合成酶共表达)小胶质细胞数量增减少,M2型(Iba1与精氨酸酶1共表达)小胶质细胞数量增多,磷酸化糖原合酶激酶3β、磷酸化β-catenin蛋白表达降低;与人参皂苷Rb1组相比,人参皂苷Rb1+抑制剂组小鼠神经功能缺损症状、脑梗死灶及脑组织水肿程度、氧化应激与炎症反应均加重,M1型小胶质细胞数量增加,M2型小胶质细胞数量减少,磷酸化糖原合酶激酶3β、磷酸化β-catenin蛋白表达升高。②结果表明,人参皂苷Rb1可调控小胶质细胞向M2型极化、缓解脑BACKGROUND:Previous studies by the research team have shown that the neuroprotective effect of ginsenoside Rb1 on improving cerebral ischemiareperfusion injury may be related to the Wnt/β-catenin signaling pathway,but the specific mechanism of action remains unclear.OBJECTIVE:To explore the molecular mechanism of ginsenoside Rb1 in alleviating cerebral ischemia-reperfusion injury in mice.METHODS:100 C57BL/6 mice were randomly divided into four groups.The sham operation group(n=25)did not undergo model establishment.In the cerebral ischemia-reperfusion injury group(n=25),the middle cerebral artery occlusion model was established by thread embolism.In the ginsenoside Rb1 group(n=25),the middle cerebral artery occlusion model was established by thread embolism and ginsenoside Rb1 was intraperitoneally injected every day for 3 days before the establishment.In ginsenoside Rb1+inhibitor group(n=25),the middle cerebral artery occlusion model was established by thread embolism and ginsenoside Rb1 and Wnt/β-catenin signaling pathway inhibitor XAV939 were intraperitoneally injected every day for 3 days before the establishment.Three days after modeling,Zea Longa score and balance beam test were used to evaluate the neurological deficits of mice.TTC staining was used to observe the volume of cerebral infarction.The dry-wet mass method was used to detect the degree of brain edema in mice.The activities of superoxide dismutase and glutathione peroxidase and the concentration of malondialdehyde in the parietal lobe of the ischemic side were detected by colorimetry.The co-expression of microglial marker Iba1 and inducible nitric oxide synthase(or arginase 1)was detected by immunofluorescence.The levels of aquaporin AQP4,inflammatory-related factors,and phosphorylation of Wnt/β-catenin pathway proteins glycogen synthase kinase 3βandβ-catenin were detected by western blot assay.The mRNA expression of inflammatory factors was detected by q-PCR.RESULTS AND CONCLUSION:(1)Compared with cerebral ischemia-reperfusion injury group,

关 键 词：脑缺血再灌注损伤 人参皂苷RB1 小胶质细胞 WNT/Β-CATENIN信号通路 神经炎症 氧化应激 神经保护 工程化组织构建 

分 类 号：R459.9[医药卫生—治疗学] R319[医药卫生—临床医学] R285.5