Protective Effects of 3-N-Butylphthalide against Oxygen-Glucose Deprivation-Induced Oxidative Damage in Astrocytes

doi:10.3969/j.issn.1673-5765.2025.08.009

Abstract

Abstract: Objective To explore the protective effects and regulatory mechanisms of 3-N-butylphthalide (NBP) against oxidative stress in astrocytes subjected to oxygen-glucose deprivation.
Methods The oxygen-glucose deprivation model was established using CTX TNA2 rat astrocytes. Cell viability was assessed using the cell counting kit-8. Superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were measured to evaluate oxidative stress levels. Real time fluorogenic quantitative PCR was used to detect the changes in the mRNA expression of pyroptosis-related genes [cysteine aspartic acid specific protease (caspase)-1, nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3), IL-1β, and IL-18]. Additionally, the expression changes of genes associated with the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway [Nrf2 and its downstream molecule heme oxygenase (HO)-1], were analyzed to explore their regulatory mechanisms.
Results Compared with the control group, the oxygen-glucose deprivation group exhibited significantly reduced cell viability (P=0.009) and SOD activity (P=0.013), alongside significantly elevated MDA content (P=0.001). The mRNA levels of caspase-1 (P=0.018), NLRP3 (P=0.021), IL-1β (P=0.004), and IL-18 (P<0.001) were significantly increased, whereas Nrf2 (P=0.035) and HO-1 (P=0.023) expression was significantly decreased. Compared with the oxygen-glucose deprivation group, the NBP-treated group demonstrated significantly increased cell viability (P=0.015) and SOD activity (P=0.013), with decreased MDA content (P=0.002). Concurrently, mRNA levels of caspase-1 (P=0.002), NLRP3 (P=0.009), IL-1β (P=0.004), and IL-18 (P=0.004) decreased, while Nrf2 (P=0.027) and HO-1 (P=0.001) expression increased.
Conclusions NBP can ameliorate oxygen-glucose deprivation-induced oxidative damage in astrocytes, potentially by activating the Nrf2 antioxidant pathway and inhibiting pyroptosis signaling, thereby alleviating oxygen-glucose deprivation-induced oxidative stress damage, and exerting a potential cytoprotective effects.

Key words: Oxygen-glucose deprivation; 3-N-butylphthalide; Oxidative stress; Pyroptosis; Nuclear factor erythroid 2-related factor 2

摘要： 目的探讨丁苯酞（3-N-butylphthalide，NBP）对氧糖剥夺后星形胶质细胞氧化应激的保护作用及调节机制。
方法采用CTX TNA2大鼠星形胶质细胞构建氧糖剥夺模型，通过细胞计数试剂盒-8型评估细胞活性，检测超氧化物歧化酶（superoxide dismutase，SOD）活性与丙二醛（malondialdehyde，MDA）含量以评估氧化应激水平；使用实时荧光定量PCR检测细胞焦亡相关基因[胱天蛋白酶（cysteine aspartic acid specific protease，caspase）-1、核苷酸结合结构域富含亮氨酸重复序列和含热蛋白结构域受体3（nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3，NLRP3）、IL-1β、IL-18]的mRNA表达变化，并分析核转录因子红系2相关因子2（nuclear factor erythroid 2-related factor 2，Nrf2）信号通路相关基因Nrf2及其下游基因血红素加氧酶（heme oxygenase，HO）-1的表达变化，以探讨其调控机制。
结果与对照组相比，氧糖剥夺组细胞活性（P=0.009）和SOD活性（P=0.013）显著降低，MDA含量显著升高（P=0.001）；caspase-1（P=0.018）、NLRP3（P=0.021）、IL-1β（P=0.004）及IL-18（P<0.001）mRNA水平显著升高，Nrf2（P=0.035）及HO-1（P=0.023）表达显著降低。与氧糖剥夺组相比，NBP组细胞活性（P=0.015）和SOD活性（P=0.013）显著升高，MDA含量（P=0.002）降低；同时，caspase-1（P=0.002）、NLRP3（P=0.009）、IL-1β（P=0.004）及IL-18（P=0.004）mRNA水平降低，Nrf2（P=0.027）及HO-1（P=0.001）基因表达升高。
结论 NBP可改善氧糖剥夺导致的星形胶质细胞氧化损伤，其可能通过激活Nrf2抗氧化通路、抑制焦亡信号，缓解氧糖剥夺诱导的氧化应激损伤，从而发挥潜在的细胞保护作用。

关键词: 氧糖剥夺; 丁苯酞; 氧化应激; 焦亡; 核转录因子红系2相关因子2

CLC Number:

GAO Yutian, LIU Wenjuan, JIN Yuling, WANG Kejian. Protective Effects of 3-N-Butylphthalide against Oxygen-Glucose Deprivation-Induced Oxidative Damage in Astrocytes[J]. Chinese Journal of Stroke, 2025, 20(8): 1006-1012.

高雨田, 刘文娟, 金玉玲, 王克健. 丁苯酞对氧糖剥夺后星形胶质细胞氧化损伤的保护作用研究[J]. 中国卒中杂志, 2025, 20(8): 1006-1012.

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