迷迭香酸对氧化应激所致神经元损伤的作用及机制  

Effect of rosmarinic acid on neuronal injury induced by oxidative stress and related mechanisms

作　　者：郭志豪[1] 赵子慧 刘捷 田林强 任文杰 GUO Zhihao;ZHAO Zihui;LIU Jie;TIAN Linqiang;REN Wenjie(Department of Orthopedics Surgery,the Third Affiliated Hospital of Xinxiang Medical University,Xinxiang 453003,Henan Province,China;Henan Key Medical Laboratory of Traumatics and Orthopaedics′Research,Xinxiang Medical University,Xinxiang 453003,Henan Province,China)

机构地区：[1]新乡医学院第三附属医院骨科,河南新乡453003 [2]新乡医学院河南省创伤与骨科研究医学重点实验室,河南新乡453003

出　　处：《新乡医学院学报》2025年第5期357-363,共7页Journal of Xinxiang Medical University

基　　金：新乡医学院第三附属医院开放课题(编号:KFKTYB202110;KFKTZD202104)。

摘　　要：目的探讨迷迭香酸(RA)对过氧化氢(H_(2)O_(2))诱导的神经元损伤的作用及其机制。方法6~8周龄无特定病原级雄性Sprauge Dawley大鼠6只,采用颈椎脱臼法处死,无菌条件下分离股骨和胫骨,提取原代骨髓间充质干细胞并诱导分化为神经元,随机分为正常组、模型组和RA干预组,正常组神经元加入完全培养基培养12 h后更换完全培养基继续培养24 h,模型组神经元加入含40μmol·L^(-1)H_(2)O_(2)的完全培养基培养12 h后更换完全培养基继续培养24 h,RA干预组神经元先加入含40μmol·L^(-1)H_(2)O_(2)的完全培养基培养12 h后再加入含25.0 mg·L^(-1)RA的完全培养基继续培养24 h,采用分光光度计法检测各组神经元中超氧化物歧化酶(SOD)活性、丙二醛(MDA)水平、谷胱甘肽(GSH)水平,实时荧光定量聚合酶链式反应法检测各组神经元中Kelch样环氧氯丙烷相关蛋白1(Keap1)、核因子红细胞系-2相关因子2(Nrf2)、还原型烟酰胺腺嘌呤二核苷酸磷酸醌氧化还原酶1(NQO1)mRNA的表达,Western blot法检测各组神经元中Keap1、Nrf2、NQO1蛋白的表达。结果模型组神经元中SOD活性、GSH水平显著低于正常组(t=12.010、37.968,P<0.01),MDA水平显著高于正常组(t=54.537,P<0.01)。RA干预组神经元中SOD活性、GSH水平显著高于模型组(t=5.645、20.926,P<0.01),MDA水平显著低于模型组(t=37.384,P<0.01)。模型组神经元中Keap1、Nrf2、NQO1 mRNA相对表达量均显著低于正常组(t=11.030、13.807、3.008,P<0.05)。RA干预组神经元中Keap1、Nrf2、NOQ1 mRNA相对表达量显著高于模型组(t=6.923、7.304、4.513,P<0.05)。模型组神经元中Keap1、Nrf2、NQO1蛋白相对表达量均显著低于正常组(t=21.787、33.864、17.069,P<0.05)。RA干预组神经元中Keap1、Nrf2、NOQ1蛋白相对表达量均显著高于模型组(t=20.452、60.093、194.183,P<0.05)。结论RA能够激活Nrf2/Keap1/抗氧化反应元件通路,保护神经元,减少H_(2)O_(2)造成的损伤。Objective To investigate the effect of rosmarinic acid(RA)on hydrogen peroxide(H_(2)O_(2))-induced neuronal injury and its mechanisms.Methods Six 6-to-8-week-old male Sprague-Dawley rats of specific pathogen-free grade were euthanized via cervical dislocation.The femurs and tibias were isolated under aseptic conditions,and primary bone marrow mesenchymal stem cells were extracted and induced to differentiate into neurons.These neurons were divided into a normal group,a model group,and a RA intervention group.Neurons in the normal group were first cultured in a complete medium for 12 hours and then in a fresh complete medium for additional 24 hours.Neurons in the model group were first cultured in a complete medium containing 40μmol·L^(-1)H_(2)O_(2)for 12 hours and then in a fresh complete medium for 24 hours.Neurons in the RA intervention group were first incubated in a complete medium containing 40μmol·L^(-1)H_(2)O_(2)for 12 hours,followed by 24 hours of cultivation in a complete medium containing 25.0 mg·L^(-1)RA.The activity of superoxide dismutase(SOD),the level of malondialdehyde(MDA),and the level of glutathione(GSH)in each group were measured using spectrophotometry.The mRNA expression levels of Kelch-like epichlorohydrin-associated protein 1(KEAP1),nuclear factor erythroid 2-related factor 2(Nrf2),and nicotinamide adenine dinucleotide phosphate quinone dehydrogenase 1(NQO1)were analyzed by real-time quantitative polymerase chain reaction.The protein expression levels of KEAP1,Nrf2,and NQO1 were detected by Western blot analysis.Results Compared with those in the normal group,the SOD activity and GSH level were significantly decreased(t=12.010,37.968;P<0.01)while the MDA level was significantly increased in the model group(t=54.537,P<0.01).The SOD activity and GSH level(t=5.645,20.926;P<0.01)were significantly higher while the MDA level was significantly lower in the RA intervention group(t=37.384,P<0.01)than those in the model group.The relative mRNA expression levels of KEAP1,Nrf2,and NQO1 in the mo

关 键 词：迷迭香酸 活性氧 Keap1-Nrf2-ARE通路 脊髓损伤 

分 类 号：R741[医药卫生—神经病学与精神病学]