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目的:探讨栝楼桂枝颗粒对脑缺血再灌注损伤大鼠氧化应激的保护作用及分子机制。方法:建立大鼠脑缺血再灌注损伤模型,栝楼桂枝颗粒灌胃给药后,核磁共振成像(magnetic resonance imaging,MRI)检测大鼠脑梗死面积,化学荧光法测定脑组织活性氧(reactive oxygen species,ROS)水平,硫代巴比妥酸法(TBA)测定丙二醛(malondialdehyde,MDA)含量,水溶性四唑蓝法(WST)测定超氧化物歧化酶(superoxide dismutase,SOD)活性,紫外法测定过氧化氢酶(catalase,CAT)和比色法测定谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活力;蛋白免疫印迹法(Western blot)和实时荧光定量PCR法(Realtime PCR)分别检测脑组织中核转录因子-E2相关因子2(NF-E2-related factor 2,Nrf2),细胞质中血红素加氧酶-1(heme oxygenase-1,HO-1),醌氧化还原酶1[NAD(P)H:quinone oxidoreductase 1,NQO1]及Kelch样ECH联合蛋白1(Kelch-like ECHassociated protein1,Keap1)的蛋白表达和mRNA相对含量。结果:栝楼桂枝颗粒能够明显减小大鼠脑梗死面积,减少ROS和MDA的产生(P<0.01),增加SOD,CAT和GSH-Px活力(P<0.01)。此外,栝楼桂枝颗粒能够上调脑组织中Nrf2,HO-1,NQO1及Keap1的表达。结论:栝楼桂枝颗粒可能通过上调Nrf2信号通路,从而抑制脑缺血再灌注损伤引起的氧化应激反应。
Objective: To investigate the protective effect and mechanism of Tielou Guizhi Granule on oxidative stress in rats with cerebral ischemia-reperfusion injury. Methods: The model of cerebral ischemia-reperfusion injury was established in rats. After the Jiaoluizhilizhi granule was administered intragastrically, the area of cerebral infarction was detected by magnetic resonance imaging (MRI). The levels of reactive oxygen species (reactive oxygen species, ROS) were detected by enzyme-linked immunosorbent assay (ELISA), malondialdehyde (MDA) content was measured by thiobarbituric acid method and superoxide dismutase (SOD) Activity and ultraviolet (UV) method were used to measure catalase (CAT) and glutathione peroxidase (GSH-Px) activity by colorimetry. Western blot and real-time fluorescence quantitative PCR (Realtime PCR) were used to detect the expression of NF-E2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1) The protein expression and mRNA relative content of oxidoreductase 1 [NAD (P) H: quinone oxidoreductase 1, NQO1] and Kelch-like ECHassociated protein1 (Keap1) Results: Guizhu Guizhi Granule can obviously reduce the area of cerebral infarction, reduce the production of ROS and MDA (P <0.01) and increase the activity of SOD, CAT and GSH-Px (P <0.01). In addition, Gu Lou Gui Zhi particles can upregulate Nrf2, HO-1, NQO1 and Keap1 expression in brain tissue. Conclusion: Guizhi Guizhi granule may inhibit oxidative stress induced by cerebral ischemia-reperfusion injury by up-regulating Nrf2 signaling pathway.