丹参酮IIA上调Nrf2/HO-1信号通路减轻海马神经元氧糖剥夺/复糖复氧损伤  

Tanshinone IIA Alleviates Oxygen-glucose Deprivation/Reoxygenation Injury of Hippocampal Neurons by Up-regulation Nrf2/HO-1 Signaling Pathway

作　　者：耿银萍 程志杰 郝艳超 王琳 GENG Yin-ping;CHENG Zhi-jie;HAO Yan-chao;WANG Lin(Department of Rehabilitation Medicine,Handan Hangang Hospital,Handan 056000,China;Department of Emergency Internal Medicine,Handan Central Hospital,Handan 056000,China;Department of Anesthesiology,Handan Hangang Hospital,Handan 056000,China)

机构地区：[1]河北省邯郸市中心医院康复医学科,邯郸056000 [2]河北省邯郸市中心医院急诊内科,邯郸056000 [3]河北省邯郸市邯钢医院麻醉科,邯郸056000

出　　处：《微循环学杂志》2024年第2期12-18,共7页Chinese Journal of Microcirculation

基　　金：河北省医学科学研究课题(20220523)。

摘　　要：目的:探讨丹参酮IIA(Tan IIA)对海马神经元氧糖剥夺/复糖复氧(OGD/R)损伤以及核因子E2相关因子2/血红素加氧酶1(Nrf2/HO-1)信号通路的影响。方法:体外培养并取对数生长期小鼠海马神经元HT22细胞开展实验,设对照(Control)组、模型(OGD/R)组、Tan IIA(5μg/ml)组、Nrf2激动剂叔丁基对苯二酚(TBHQ,1.6μg/ml)组和Tan IIA(5μg/ml)+TBHQ(1.6μg/ml)组。各组分别给药干预2h后,除Control组外,其它组通过氧糖剥夺6h后复糖复氧24h构建HT22细胞OGD/R损伤模型。MTT法检测细胞活力,流式细胞术检测细胞凋亡率,二氯荧光素双乙酸盐(DCFH-DA)探针法检测细胞中活性氧(ROS)含量,硫代巴比妥酸法检测丙二醛(MDA)含量,黄嘌呤氧化法、钼酸铵法检测超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性,实时荧光定量聚合酶链式反应(RT-PCR)和Western Blotting检测Nrf2/HO-1信号通路相关mRNA和蛋白表达。结果:与OGD/R组相比,Tan IIA组、TBHQ组和Tan IIA+TBHQ组HT22细胞活力明显升高、凋亡率明显降低(P<0.05);细胞中ROS、MDA含量明显降低,SOD、CAT活性明显升高(P<0.05);Nrf2、HO-1 mRNA和蛋白表达量明显升高,B淋巴细胞瘤2(Bcl-2)蛋白表达量明显升高,Bcl-2相关X蛋白(Bax)、激活型半胱氨酸蛋白酶-3(Cleaved Caspase-3)蛋白表达量及Bax/Bcl-2、Cleaved Caspase-3/Caspase-3比值明显降低(P<0.05)。Tan IIA+TBHQ组对各检测指标的调控作用明显优于Tan IIA组和BHQ组(P<0.05)。结论:Tan IIA对OGD/R损伤海马神经元氧化应激损伤和凋亡具有抑制作用,其机制可能与上调Nrf2/HO-1信号通路有关。Objective:To investigate the effects of tanshinone IIA(Tan IIA)on oxygen-glucose deprivation/reoxygenation(OGD/R)injury and nuclear factor E2-related factor 2/heme oxygenase-1(Nrf2/HO-1)signaling pathway in hippocampal neurons.Method:HT22 cells of mouse hippocampal neurons were cultured in vitro and which in logarithmic growth period was used as experimental cell.And the Control group,model(OGD/R)group,Tan IIA(5μg/ml)group,Nrf2 agonist tert-butylhydroquinone(TBHQ,1.6μg/ml)group and Tan IIA(5μg/ml)+TBHQ(1.6μg/ml)group were set up.After 2h of treatment in each group,except for the Control group,the OGD/R damaged HT22 cell model was prepared by reoxygenation for 24h after 6h of oxygen-glucose deprivation.Cell viability was detected by MTT method,cell apoptosis rate was detected by flow cytometry,reactive oxygen species(ROS)content was detected by dichlorofluorescein diacetate(DCFH-DA)probe method,malondialdehyde(MDA)content was detected by thiobarbituric acid method,and superoxide dismutase(SOD)and catalase(CAT)activities were detected by xanthine oxidation method and ammonium molybdate method.Real-time fluorescent quantitative polymerase chain reaction(RT-PCR)and Western Blotting were used to detect mRNA and protein expression related to Nrf2/HO-1 signaling pathway.Results:Compared with the OGD/R group,the activity of HT22 cells in Tan IIA group,TBHQ group and Tan IIA+TBHQ group were significantly increased,and the apoptosis rate was significantly decreased(P<0.05).The content of ROS,MDA were significantly decreased,and the activity of SOD,CAT were significantly increased(P<0.05).The mRNA and protein expression of Nrf2,HO-1 were significantly increased,the protein expression of B lymphoblastoma 2(Bcl-2)was significantly increased,the protein expression of Bcl-2 associated X protein(Bax),Cleaved Caspase-3 and the ratio of Bax/Bcl-2,Cleaved Caspase-3/Caspase-3 were significantly decreased(P<0.05).Tan IIA+TBHQ group had significantly better regulation on the detection indicators than those of Tan IIA group and BHQ

关 键 词：海马神经元 丹参酮IIA 氧糖剥夺/复糖复氧 Nrf2/HO-1信号通路 氧化应激 凋亡 

分 类 号：R743.3[医药卫生—神经病学与精神病学]