The protective effect of SOX5 on ischemic stroke through Nrf2/HO-1 signaling pathway

doi:10.19854/j.cnki.1008-2425.2021.05.0004

Abstract

Abstract: Objective To explore the neuroprotective effect of transcription factor SOX5 on ischemic stroke rats and its effect on nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway. Methods Thirty-six rats were randomly divided into sham operation group,model group and overexpression group,each with 12 rats.A rat model of middle cerebral artery occlusion (MCAO) ischemic stroke was prepared by suture method.One hour before modeling,1μl of sterile normal saline,10μl of blank negative control virus vector,and 10μl of SOX5 overexpressed lentiviral vector were injected into the lateral ventricle of rats in the sham operation group,model group and overexpression group with a micro-injector.Longa 5-level scoring method was used to evaluate neural function;TUNEL method was used to detect cell apoptosis;Enzyme-linked immunosorbent assay to detect the expression of reactive oxygen species (ROS),malondialdehyde (MDA),glutathione (GSH),interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α) in brain tissue Level;Western-blot detects the expression of Nrf2 and HO-1. Results There was no cerebral infarction in rats in the sham operation group.The percentage of cerebral infarction volume in the model group was (42.09±9.34)%,which was higher than (19.34±6.21)% in the overexpression group (P＜0.05).At 24h and 48h of reperfusion,the neurological scores of the model group were higher than those of the sham operation group and the overexpression group (P＜0.05).There was no obvious apoptosis in the sham operation group.The apoptosis rate of the model group was (44.39±3.27)%,which was higher than that of the overexpression group (17.34±4.39)% significant (P＜0.05).Compared with the sham operation group,the expression level of ROS,MDA,IL-1β and TNF-α in the model group was higher,while the expression level of GSH was lower (P＜0.05).Compared with the model group,the expression level of ROS,MDA,IL-1β and TNF-α in the overexpression group was lower,while the expression level of GSH was higher (P＜0.05).The nuclear-to-cytoplasmic ratio of Nrf2 protein and the expression level of HO-1 protein in the model group were higher than those in the sham operation group (P＜0.05).The nuclear-to-cytoplasmic ratio of Nrf2 protein and the expression level of HO-1 protein in the overexpression group were also higher than those in the model group (P＜0.05). Conclusion SOX5 may exert anti-oxidative stress and anti-inflammatory effects through the Nrf2/HO-1 signaling pathway,thereby improving the neurological function of rats with ischemic stroke.

Key words: Transcription factor SOX5, Nuclear factor E2 related factor 2, Heme oxygenase-1, Ischemic stroke, Oxidative stress, Inflammation

摘要： 目的探讨转录因子SOX5对缺血性脑卒中大鼠的神经保护作用及对核因子E2相关因子2(Nrf2)/血红素加氧酶-1(HO-1)通路的影响。方法将36只大鼠随机分为假手术组、模型组和过表达组,各12只。用线栓法制备大脑中动脉闭塞(MCAO)缺血性脑卒中大鼠模型。造模前1h,用微量进样器将1μl无菌生理盐水、10μl 空白阴性对照病毒载体、10μl SOX5过表达慢病毒载体,分别注入假手术组、模型组和过表达组大鼠的侧脑室。用Longa 5级评分法评估神经功能;TUNEL法检测在体细胞凋亡;酶联免疫吸附实验检测脑组织中活性氧(ROS)、丙二醛(MDA)、谷胱甘肽(GSH)、白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)表达水平;Western-blot检测Nrf2和HO-1表达。结果假手术组大鼠未见脑梗死,模型组脑梗死体积百分比为(42.09±9.34)%,高于过表达组的(19.34±6.21)%,差异有统计学意义(P＜0.05)。再灌注24h和48h时,模型组神经功能评分均高于假手术组和过表达组(P＜0.05)。假手术组未见明显细胞凋亡。模型组细胞凋亡率为(44.39±3.27)%,高于过表达组的(17.34±4.39)%,差异有统计学意义(P＜0.05)。与假手术组比较,模型组ROS、MDA、IL-1β和TNF-α表达水平较高,而GSH表达水平较低(P＜0.05)。与模型组比较,过表达组ROS、MDA、IL-1β和TNF-α表达水平较低,而GSH表达水平较高(P＜0.05)。模型组Nrf2蛋白的核质比、HO-1蛋白表达水平均高于假手术组(P＜0.05)。过表达组Nrf2蛋白的核质比、HO-1蛋白表达水平也均高于模型组(P＜0.05)。结论 SOX5可能通过Nrf2/HO-1信号通路发挥抗氧化应激和抗炎作用,进而改善缺血性脑卒中大鼠的神经功能。

关键词: 转录因子SOX5, 核因子E2相关因子2, 血红素加氧酶-1, 缺血性脑卒中, 氧化应激, 炎症

CLC Number:

R743

Li Jianhong, Zhang Yuhui, Wang Bufei. The protective effect of SOX5 on ischemic stroke through Nrf2/HO-1 signaling pathway[J]. Chinese Journal of Stereotactic and Functional Neurosurgery, 2021, 34(5): 274-278.

李建红, 张余辉, 王布飞. SOX5通过Nrf2/HO-1信号通路对缺血性脑卒中的保护作用[J]. 立体定向和功能性神经外科杂志, 2021, 34(5): 274-278.

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