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纸质出版日期:2018
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胡跃强, 秦红玲, 唐农, 等. 基于Nrf2/ARE信号通路探讨清热化瘀方对大鼠脑缺血再灌注损伤氧化应激反应的影响[J]. 中国实验方剂学杂志, 2018,24(14):128-133.
HU Yue-qiang, QIN Hong-ling, TANG Nong, et al. Qingre Huayu Prescription Reduces Oxidative Stress in Rats with Cerebral Ischemia-reperfusion Injury Through Activating Nrf2/ARE Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(14): 128-133.
胡跃强, 秦红玲, 唐农, 等. 基于Nrf2/ARE信号通路探讨清热化瘀方对大鼠脑缺血再灌注损伤氧化应激反应的影响[J]. 中国实验方剂学杂志, 2018,24(14):128-133. DOI: 10.13422/j.cnki.syfjx.20181329.
HU Yue-qiang, QIN Hong-ling, TANG Nong, et al. Qingre Huayu Prescription Reduces Oxidative Stress in Rats with Cerebral Ischemia-reperfusion Injury Through Activating Nrf2/ARE Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(14): 128-133. DOI: 10.13422/j.cnki.syfjx.20181329.
目的:从核转录因子E2相关因子2/抗氧化反应元件(Nrf2/ARE)信号通路探讨清热化瘀方对脑缺血再灌注损伤大鼠氧化应激的影响及可能的作用机制。方法:采用Longa线栓法制备局灶性脑缺血再灌注大鼠(MCAO)模型,将160只SD大鼠随机分为4组,分别为假手术组(SO),模型组(MCAO),清开灵组(QKL),清热化瘀方组(QRHY)。每组大鼠40只,根据取材时间点12 h,1 d,2 d,3 d可以分为每组分为4个亚组,每个亚组10只大鼠。分别采用氯化三苯基四氮唑(TTC)染色法测量脑梗死体积,实时荧光定量聚合酶链式反应(Real-time PCR)和免疫组化检测Nrf2/ARE信号通路蛋白Nrf2和血红素加氧酶1(HO-1) mRNA及其蛋白表达变化。结果:与SO组比较,大鼠脑缺血再灌注后1 d,MCAO组梗死体积显著升高(P<0.01),QKL组较MCAO组梗死体积明显减小(P<0.05);QRHY组脑梗死体积较QKL组进一步缩小(P<0.05);MCAO组Nrf2和HO-1 mRNA及其蛋白表达均于缺血再灌注后12 h开始明显上升,24 h达高峰(P<0.05,P<0.01),随再灌注时间延长其表达逐渐下降,但仍保持较高表达水平(P<0.05);QKL组较MCAO组Nrf2和HO-1 mRNA及其蛋白表达明显升高(P<0.05),QRHY组较QKL组进一步升高Nrf2和HO-1 mRNA及其蛋白表达(P<0.05,P<0.01)。结论:清热化瘀方治疗脑缺血再灌注损伤可能通过激活Nrf2/ARE信号通路而降低细胞氧化损伤的程度,从而达到保护神经元的作用。
Objective: To investigate the effect and possible mechanism of Qingre Huayu prescription on oxidative stress in rats with cerebral ischemia-reperfusion injury (I/R) through nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) pathway. Method: The focal cerebral ischemia rat (MCAO) model was prepared by suture method
and 160 SD rats were randomly divided into 4 groups:sham operation group (SO)
model group (MCAO)
Qingkailing control group (QKL)
and Qingre Huayu prescription group (QRHY)
n=40 in each group. Each group was further divided into 4 subgroups according to 12 h
1 d
2 d and 3 d after I/R
n=10 in each sub group. Cerebral infarction volume was tested by triphenyltetrazolium chlorid(TTC) staining method
and the changes of the mRNA and protein expression of Nrf2 and heme oxygenase 1 (HO-1) at each time point after reperfusion were observed by real-time fluorescence quantitative PCR(Real-time PCR) and immunohistochemistry separately. Result: As compared with the normal group
the brain infarct volume was significantly increased in MCAO group 1 d after cerebral ischemia reperfusion (P<0.01); the brain infarct volume in QKL group was significantly smaller than that of MCAO group (P<0.05
P<0.01); QRHY could further decrease the brain infarct volume as compared with QKL group (P<0.05). The mRNA and protein expression levels of Nrf2 and HO-1 in MCAO group were increased significantly 12 h after I/R
and reached the peak at 24 h (P<0.05
P<0.01)
then decreased continuously with the increase of I/R time
but still maintained high expression levels (P<0.05). QKL could increase the mRNA and protein expression levels of Nrf2 and HO-1 as compared with MCAO group (P<0.05). QRHY could further increase the above expression as compared with QKL group (P<0.05). Conclusion: QRHY may up-regulate the expressions of Nrf2 and HO-1 in rats and relieve I/R through activating the Nrf2/ARE signaling pathway.
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