摘要
目的探究灯盏花素(Bre)对大鼠模型中颅内动脉瘤(IA)的形成和对核因子E2相关因子2(Nrf2)途径的影响。方法通过弹性蛋白酶注射建立大鼠IA模型,将大鼠随机分为假手术组、模型组、Bre组,每组15只。Bre组每天腹腔注射50 mg/kg Bre,假手术组和模型组腹腔注射等体积生理盐水,持续3周,在此期间记录IA的发生率、生存率和收缩压,免疫荧光染色和qRT-PCR检测α-平滑肌肌动蛋白(α-SMA)和平滑肌22α(SM22α)的表达。使用过氧化氢(H_(2)O_(2),0.5 mmol/L)处理大鼠血管平滑肌细胞(VSMC)诱导氧化损伤,然后与Bre(100μmol/L)或(和)ML385(Nrf2抑制剂)共孵育,Western blot、qRT-PCR、ELISA、DCFH-DA荧光染色、流式细胞术分别检测Nrf2、收缩表型相关蛋白和炎性细胞因子的表达、活性氧的产生和细胞凋亡率。结果在IA大鼠中,Bre处理上调核Nrf2的表达,改善IA病理变化,降低IA的发生率,改善生存率,并降低收缩压。在H_(2)O_(2)处理的VSMC中,Bre预处理升高Nrf2、抗氧化酶、α-SMA和SM22α的表达,降低基质金属蛋白酶(MMP)-2和MMP-9表达、活性氧的产生和细胞凋亡,减轻脑动脉中炎性细胞浸润和促炎性细胞因子的表达。抑制Nrf2减弱了Bre预处理在H_(2)O_(2)处理的VSMC中的治疗效果。结论Bre可有效降低VSMC中的氧化应激和炎症反应,从而减轻大鼠中IA的形成和破裂,其机制可能与Nrf2途径的激活有关。
Objective To explore the effect and mechanism of breviscapine(Bre)on the formation of intracranial aneurysm(IA)and its influence on nuclear factor erythroid 2-related factor 2(Nrf2)pathway in a rat model.Methods The rat model of IA was established by elastase injection.The rats were randomly divided into Sham group,Model group and Bre group with 15 rats in each group.The Bre group was intraperitoneally injected with 50 mg/kg Bre every day,the Sham group and Model group were intraperitoneally injected with the same volume of normal saline for 3 weeks.During this period,the incidence of IA,survival rate and systolic blood pressure were recorded,immunofluorescence staining and qRT-PCR were employed to assay the expression ofα-smooth muscle actin(α-SMA)and smooth muscle 22α(SM22α).Hydrogen peroxide(H_(2)O_(2),0.5 mmol/L)was used to treat rat vascular smooth muscle cells(VSMC)to induce oxidative damage,and then co-incubated with Bre(100μmol/L)or/and ML385(Nrf2 inhibitor),Western blot,qRT-PCR,ELISA,DCFH-DA fluorescence staining and flow cytometry were used to detect the expression of Nrf2,contractile phenotype-related protein and inflammatory cytokine,reactive oxygen species(ROS)production and cell apoptosis rate,respectively.Results In IA rats,Bre treatment up-regulated the expression of nuclear Nrf2,improved the pathological changes of IA,reduced the incidence of IA,improved the survival rate,and lowered the systolic blood pressure.In VSMC treated with H_(2)O_(2),Bre pretreatment increased the expression of Nrf2,antioxidant enzymes,α-SMA and SM22α,reduced the expression of matrix metalloproteinases(MMP)-2 and MMP-9,the production of ROS and cell apoptosis,and reduced inflammation in cerebral arteries cell infiltration and expression of pro-inflammatory cytokines.Inhibition of Nrf2 weakened the therapeutic effect of Bre pretreatment in H_(2)O_(2)treated VSMC.Conclusion Bre can effectively reduce the oxidative stress and inflammation in VSMC,thereby reducing the formation and rupture of IA in rat.The mechanism m
作者
艾奇渊
王勇
徐瑞春
彭臻
李劲松
Ai Qiyuan;Wang Yong;Xu Ruichun;Peng Zhen;Li Jinsong(Dept of Neurosurgery,The Third Affiliated Hospital of Guizhou Medical University,Duyun 558000)
出处
《安徽医科大学学报》
CAS
北大核心
2022年第4期579-585,共7页
Acta Universitatis Medicinalis Anhui
基金
贵州省科技合作计划(编号:黔科合LH字[2019]7162号)。
