莱菔硫烷对Nrf2原代星形胶质细胞Ⅱ相酶的诱导作用  

Induction of the phase Ⅱ enzyme of Nrf2 primary astrocytes by sulforaphane

作　　者：温迪[1] 于秀军[1] 郭艳苏[1] 陈相[1] 李春岩[1] Wen Di;Yu Xiujun;Guo Yansu;Chen Xiang;Li Chunyan(Department of Neurology,the Second Hospital of Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]河北医科大学第二医院神经内科,石家庄050000

出　　处：《脑与神经疾病杂志》2022年第6期361-365,共5页Journal of Brain and Nervous Diseases

基　　金：河北省自然科学基金青年科学基金项目(H2021206048)。

摘　　要：目的 探索莱菔硫烷(SFN)对培养的Nrf2+/+、Nrf2-/-原代星形胶质细胞Ⅱ相酶的诱导作用。方法 取经基因鉴定的Nrf2+/+、Nrf2-/-新生乳鼠,取出大脑皮质分离培养原代星形胶质细胞,并进行纯化与鉴定。将细胞分为Nrf2+/+和Nrf2-/-组,每组又分为溶剂对照组和SFN药物干预组。通过给予不同浓度的SFN干预Nrf2+/+组细胞,测定Ⅱ相酶的表达来选择最佳给药浓度。然后分别给予Nrf2+/+、Nrf2-/-组DMSO和最适浓度的SFN,通过Western blot测定Ⅱ相酶的表达。结果 不同浓度SFN干预Nrf2+/+星形胶质细胞显示,10μmol·L^(-1)对Ⅱ相酶Gclm、Nqo1、Ho1蛋白表达的诱导作用最强,为最佳给药浓度。Nrf2+/+和Nrf2-/-细胞分别给予10μmol·L^(-1) SFN处理后,Ho1均能明显上调。Nqo1和Gc1m只在Nrf2+/+细胞中明显上调,在Nrf2-/-细胞中不上调。而对于Gss和Gclc,给药后Nrf2+/+和Nrf2-/-细胞两种酶表达均无明显变化。结论 SFN对原代培养的星形胶质细胞Ho1的诱导不完全依赖于核转录因子Nrf2;对Nqo1、Gclm的诱导很大程度上依赖于转录因子Nrf2;对Gss和Gclc的表达无明显诱导作用。Objective To explore the induction of the phase II enzymes of cultured Nrf2+/+and Nrf2-/-primary astrocytes by sulforaphane.Methods The genetically identified Nrf2+/+,Nrf2-/-newborn suckling mice were taken out,the primary astrocytes were isolated and cultured,then purified and identified.The cells were divided into 2 groups,the Nrf2+/+and Nrf2-/-groups,and each group was further divided into a solvent control group and the sulforaphane drug intervention group.The optimal concentration of the drug was selected through the expression of the phase Ⅱ enzyme by administering different concentrations of sulforaphane to the Nrf2+/+group of cells.Then,DMSO and the optimal concentration of sulforaphane were respectively administered to the Nrf2+/+,Nrf2-/-groups,and the expression of phase II enzyme was determined by western blot analysis.Results Intervention of Nrf2+/+astrocytes with different concentrations of sulforaphane showed that 10 μmol·L^(-1) had the strongest induction effect on the expression of phase II enzymes Gclm,Nqol and Ho1,which was the optimal concentration.Ho1 was significantly up-regulated in both Nrf2+/+and Nrf2-/-cells after treatment with 10 μmol·L^(-1) sulforaphane.Nqo1 and Gclm were only up-regulated only in Nrf2+/+cells and not up-regulated in Nrf2-/-cells.While there was no significant change in the expression of Gss or Gclc in Nrf2+/+or Nrf2-/-cells after administration of sulforaphane.Conclusion The induction of Ho1 was not completely dependent on the nuclear transcription factor Nrf2 in primary cultured astrocytes by sulforaphane;the induction of Nqol and Gclm was largely dependent on the transcription factor Nrf2;while no significant difference was observed in the expression of Gss or Gclc by sulforaphane.

关 键 词：原代星形胶质细胞 氧化应激 莱菔硫烷 Ⅱ相酶 NRF2 

分 类 号：R741[医药卫生—神经病学与精神病学]