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中国临床药理学与治疗学 ›› 2025, Vol. 30 ›› Issue (1): 11-19.doi: 10.12092/j.issn.1009-2501.2025.01.002

• 基础研究 • 上一篇    下一篇

Nrf1通过抑制细胞凋亡减轻氧糖剥夺/复糖复氧致神经元损伤

夏荣松1,2,杨靖2,王红1,彭哲3,赵一蓓1,杨俊卿1   

  1. 1重庆医科大学药理学系生物化学与分子药理学重点实验室,重庆  400016;2重庆市人民医院药学部,重庆  401147;3重庆医科大学附属妇女儿童医院药学部,重庆  401147

  • 收稿日期:2024-01-08 修回日期:2024-04-22 出版日期:2025-01-26 发布日期:2025-01-02
  • 通讯作者: 杨俊卿,男,二级教授,研究方向:神经精神药理。 E-mail:cqyangjq@cqmu.edu.cn
  • 作者简介:夏荣松,男,硕士研究生,研究方向:神经精神药理。 E-mail:rongsong1987@stu.cqmu.edu.cn
  • 基金资助:
    国家自然科学基金面上项目(82173793)

Nrf1 attenuates neuronal injury caused by oxygen glucose deprivation/reperfusion via inhibiting apoptosis

XIA Rongsong1,2, YANG Jing2, WANG Hong1, PENG Zhe3, ZHAO Yibei1, YANG Junqing1   

  1. 1 Key Laboratory of Biochemistry and Molecular Pharmacology, Department of Pharmacology, Chongqing Medical University, Chongqing 400016, China; 2 Department of Pharmacy, Chongqing General Hospital, Chongqing 401147, China; 3 Department of Pharmacy, Women and Children's Hospital of Chongqing Medical University, Chongqing 401147, China
  • Received:2024-01-08 Revised:2024-04-22 Online:2025-01-26 Published:2025-01-02

摘要:

目的:探讨Nrf1对氧糖剥夺/复糖复氧致神经元损伤的作用以及机制。方法:通过GEO数据库单细胞测序数据,基于GSVA包计算评价细胞Nrf1表达与凋亡通路的相关性。后续实验将PC12细胞和原代神经元分为正常组(Normal)、模型组(OGD/R)、OGD/R+siRNA-NC组及OGD/R+Nrf1-siRNA-2组,采用倒置显微镜观察细胞形态,MTT法检测细胞活力,流式细胞术检测细胞凋亡率,DHE荧光探针检测活氧性(ROS)水平,Western blot法检测Nrf1、bcl-2、Bax蛋白表达水平,激光扫描共聚焦显微镜观察细胞核转位。结果:生信分析结果发现Nrf1主要富集于凋亡通路;与Normal组相比,OGD/R处理使PC12细胞和原代神经元突触断裂、胞体变小、大量细胞聚集,存活率显著降低(P<0.01),ROS水平显著升高(P<0.01),Nrf1、Bax蛋白表达水平显著升高(P<0.05),bcl-2蛋白表达水平显著降低(P<0.05);与OGD/R组相比,siRNA-NC组细胞各项指标均无显著差异;与siRNA-NC组相比,Nrf1-siRNA-2组使PC12细胞和原代神经元突触完全丧失,细胞活力显著降低(P<0.01),ROS水平显著增加(P<0.01),Nrf1、Bax蛋白表达水平进一步上调,而bcl-2蛋白表达水平下调(P<0.05)。结论:Nrf1可通过抑制细胞凋亡减轻OGD/R诱导的神经元损伤。

关键词: OGD/R, Nrf1, Bax, bcl-2, 凋亡

Abstract:

AIM: To investigate the effects of Nrf1 on neuronal apoptosis treated by oxygen-glucose deprivation/reperfusion and the mechanism. METHODS: Single-cell sequencing data was analyzed by GEO database, and the correlation of Nrf1 expression with apoptotic pathways evaluated based on GSVA package calculations. PC12 cells and primary neurons were divided into the Normal group, the OGD/R group, the OGD/R+ siRNA-NC group, and the OGD/R+ Nrf1-siRNA-2 group. Cell images was observed by laser scanning confocal microscopy; the viability of cells were detected by MTT assay; the apoptosis of cells were detected by flow cytometry; DHE fluorescent probe to detect ROS levels, the protein expression of Nrf1,bcl-2 and Bax were detected by Western blot; and nuclear translocation was observed by laser scanning confocal microscope. RESULTS: The results of biosignature analysis revealed that Nrf1 was mainly enriched in the apoptotic pathway; compared with normal group, the cell body became smaller, synapse broke, cells clustered in PC12 cells and primary neurons with OGD/R treated, and the vitality of PC12 cells and neuronal were decreased significantly (P<0.01); ROS levels were significantly higher (P<0.01), the expressions of Nrf1 and Bax were increased significantly, and the expression of bcl-2 was decreased significantly (P<0.05). Compared with the OGD/R group, there was no significant difference in the siRNA-NC group; compared with the siRNA-NC group, the viability of cells was decreased significantly (P<0.01); ROS levels were significantly increased (P<0.01), the expressions of Nrf1 and Bax were increased markedly, and the expressions of bcl-2 was decreased significantly (P<0.05) in Nrf1-siRNA-2 group. CONCLUSION: Nrf1 attenuates neuronal injury caused by oxygen glucose deprivation/reperfusion via inhibiting apoptosis.

Key words: OGD/R, Nrf1, Bax, bcl-2, apoptosis

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