SIRT1/FoxO1信号通路在三叶苷减轻大鼠脑缺血再灌注损伤中的作用
Role of SIRT1/FoxO1 signaling pathway in trilobatin-induced reduction of cerebral ischemia-reper-fusion injury in rats
高美娜 1王磊 1丁彦玲1
作者信息
- 1. 保定市第一中心医院麻醉科,保定 071000
- 折叠
摘要
目的 评价沉默信息调节因子1(SIRT1)/叉头框蛋白01(FoxO1)信号通路在三叶苷减轻大鼠脑缺血再灌注损伤中的作用.方法 清洁级健康雄性SD大鼠80只,6～8周龄,体质量230～280 g,采用随机数字表法分为4组(n=20):假手术组(S组)、脑缺血再灌注组(CIR组)、三叶苷+脑缺血再灌注组(T组)和三叶苷+脑缺血再灌注+SIRT1/Fox01信号通路抑制剂EX527组(E组).采用大脑中动脉栓塞法制备大鼠局灶性脑缺血再灌注损伤模型.T组和E组大鼠于缺血前3 d时开始灌胃给予三叶苷15 mg/kg,2次/d,连续3 d;E组大鼠每次灌胃前腹腔注射EX527 5 mg/kg.于再灌注24 h时采用改良Longa评分评估神经功能,随后处死大鼠取全脑组织,采用TTC染色确定脑梗死体积,流式细胞术检测海马神经元凋亡率和ROS水平,Western blot法检测SIRT1及乙酰化FOXO1(Ac-FOXO1)表达,ELISA法检测SOD和MDA含量,HE染色后光镜下观察海马CA1区病理学结果.结果 与S组比较,CIR组Longa评分、脑梗死体积、海马神经元凋亡率、ROS和MDA水平升高,SOD含量降低,SIRT1表达下调,Ac-FOXO1表达上调(P＜0.05);与CIR组比较,T组Longa评分、脑梗死体积、海马神经元凋亡率、ROS和MDA水平降低,SOD含量升高,SIRT1表达上调,Ac-FOXO1表达下调(P＜0.05);与T组比较,E组Longa评分、脑梗死体积、海马神经元凋亡率、ROS和MDA水平升高,SOD含量降低,SIRT1表达下调,Ac-FOXO1表达上调(P＜0.05).结论 三叶苷可能通过激活SIRT1/FoxO1信号通路,抑制海马氧化应激反应和神经元凋亡,减轻大鼠脑缺血再灌注损伤.
Abstract
Objective To evaluate the role of the SIRT1/FoxO1 signaling pathway in trilobatin-in-duced reduction of cerebral ischemia-reperfusion(I/R)injury in rats.Methods Eighty clean-grade healthy male Sprague-Dawley rats,aged 6-8 weeks,weighing 230-280 g,were divided into 4 groups(n=20 each)using a random number table method:sham operation group(group S),cerebral I/R group(group CIR),trilobatin+cerebral I/R group(group T)and trilobatin+cerebral I/R+SIRTl/FoxO1 signaling pathway inhibitor EX527 group(group E).The model of focal cerebral I/R injury was established by middle cerebral artery occlusion in anesthetized animals.Trilobatin 15 mg/kg was given by gavage twice a day for 3 consecu-tive days starting from 3 days before ischemia in T and E groups.EX527 5 mg/kg was intraperitoneally injec-ted before each gavage in group E.Modified Longa scoring scale was used to assess neurological function at 24 h of reperfusion,then the rats were sacrificed and whole brain tissues were obtained for determination of cerebral infarct size(using TTC staining),apoptosis rate and level of reactive oxygen species(ROS)in the hippocampus(by flow cytometry),expression of SIRT1 and acetylated FOXO1(Ac-FOXO1)(by Western blot)and contents of superoxide dismutase(SOD)and malondialdehyde(MDA)(by enzyme-linked immu-nosorbent assay)and for microscopic examination of pathological changes in the hippocampal CAI area after HE staining.Results Compared with group S,Longa score,cerebral infarct size,apoptosis rate of hipp-ocampal neurons,and levels of ROS and MDA were significantly increased,the content of SOD was de-creased,the expression of SIRT1 was down-regulated,and the expression of Ac-FOXOl was up-regulated in group CIR(P＜0.05).Compared with group CIR,Longa score,cerebral infarct size,apoptosis rate of hipp-ocampal neurons,and levels of ROS and MDA were significantly decreased,the content of SOD was in-creased,the expression of SIRT1 was up-regulated,and the expression of Ac-FOXO1 was down-regulated in group T(P＜0.05).Compared with group T,Longa score,cerebral infarct size,apoptosis rate of hippocam-pal neurons,and levels of ROS and MDA were significantly increased,the content of SOD was decreased,the expression of SIRT1 was down-regulated,and the expression of Ac-FOXO1 was up-regulated in group E(P＜0.05).Conclusions Trilobatin may inhibit oxidative stress responses and neuronal apoptosis in hipp-ocampi by activating the SIRT1/FoxO1 signaling pathway,thus alleviating cerebral I/R injury in rats.
关键词
类黄酮物质/再灌注损伤/脑/抗衰老酶1/叉头框蛋白O1Key words
Flavonoids/Reperfusion injury/Brain/Sirtuin 1/Forkhead box protein O1引用本文复制引用
基金项目
河北省医学科学研究项目(20220289)
出版年
2024
NETL
NSTL
万方数据