Nrf3通路参与星形胶质细胞对抗鱼藤酮毒性的保护作用

中国生物工程杂志

2010, Vol. 30

Issue (01): 19-24

研究报告

Nrf3通路参与星形胶质细胞对抗鱼藤酮毒性的保护作用

苏玉金¹,曹倩²,鲁玲玲²,孙晓红²,高华²,赵莎莎²,杨慧²

1.首都医科大学北京市神经外科研究所北京 100069
2.首都医科大学北京神经科学研究所北京神经再生修复研究重点实验室教育部神经变性病学重点实验室北京 100069

Nrf3 Pathway was Involved in the Protection of Astrocyte Against the Neurotoxicity of Rotenone

SU Yu-jin¹,CAO Qian²,LU Ling-ling²,SUN Xiao-hong²,GAO Hua²,ZHAO Sha-sha²,YANG Hui²

1.Beijing Neurosurgical Institute,Capital Medical University,Beijing 100069,China
2.Beijing Institute for Neuroscience , Capital Medical University, Beijing Center of Neural Regeneration and Repair, Key Laboratory for Neurodegenerative Disease of the Ministry of Education, Beijing 100069, China

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摘要：

目的：寻找星形胶质细胞在对抗由鱼藤酮导致的氧化应激中发挥保护作用的相关分子并探讨其作用机制。方法：小鼠多巴胺能MN9D细胞分别在星形胶质细胞条件培养液（ACM）与星形胶质细胞用新鲜培养基中培养24小时后加入鱼藤酮作用48小时。细胞计数，评价星形胶质细胞条件培养液对MN9D细胞的保护作用。利用基因芯片技术寻找MN9D细胞在ACM处理后发生表达上调或下调基因，并对这些基因进行分析，找出有意义基因。结果：在不同作用时间和鱼藤酮浓度梯度下，经过ACM处理的MN9D细胞活性显著高于在普通培养基中培养的细胞。初步得到104个差异表达基因，其中62个表达上调基因，42个表达下调基因。这些基因主要与凋亡、肿瘤、细胞周期、代谢、信号转导、转录调节、翻译调节和传递蛋白等相关。对其中的Atp5a1，Nrf3基因进行分析，发现Nrf3通路参与了ACM的保护作用。结论： ACM能保护MN9D细胞抵抗鱼藤酮所致的细胞毒性， Atp5a1，Nrf3，GCL，NQO1等基因经ACM处理后发生差异表达，可能是星形胶质细胞保护作用的部分下游信号通路。

关键词： 帕金森病; 星形胶质细胞; Nrf3; 基因芯片

Abstract:

Objective:To find out genes that are responsibe for the protection of astrocyte against the neurotoxicity of rotenone. Methods: To address this question, MN9D cells was used as a cell model of dopaminergic neurons and rotenone a toxin to initiate mitochondrial deficiency and wonder whether ACM could protect MN9D cells against rotenone toxicity and its possible protective mechanism. Cells treated with ACM or not were exposed to rotenone before assay of cell viability，signal transduction pathway for apoptosis and so on. Results: The results showed that rotenone decreased viability of MN9D cells in a dose-dependent manner and ACM treatment significantly attenuated rotenone toxicity at each concentration. Results of genechip found 104 differentially expressed genes, among which there were 62 up-regulated genes and 42 down-regulated ones after ACM treatment. These genes involved many functions such as signal transduction, transcription regulation, metabolism, cell cycle, etc. According to previous literature and the present study, 3 genes, namely Nrf3, GCL and NQO1 were likely to be associated with ACM protection and were confirmed by real-time PCR. Conclusions:The findings suggest that astrocytes can protect MN9D cells from oxidative stress and mitochondrial dysfunction caused by rotenone. Glutathione and neurotrophic factors may partially account for the protection. Some meaningful genes related to ACM protection were selected by genechip. Further research into these genes may provide clues to the detailed mechanism of ACM protection and shed light on accurate role of astrocytes in Parkinson’s disease.

Key words: Parkinson’s disease;Astrocyte Nrf3;Gene chips

收稿日期: 2009-07-07 出版日期: 2010-01-27

基金资助:

国家“973”计划(2006CB500706)、国家自然科学基金(30670655，30970940)、国家自然科学基金委主任基金(30950003)、教育部高等学校博士学科点专项科研基金(20060025004)、北京市自然基金面上项目（7082011，5102007,5102012）资助项目

通讯作者: 杨慧 E-mail: huiyang@ccum.edu.cn

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引用本文:

苏玉金曹倩鲁玲玲孙晓红高华赵莎莎杨慧. Nrf3通路参与星形胶质细胞对抗鱼藤酮毒性的保护作用[J]. 中国生物工程杂志, 2010, 30(01): 19-24.

SU Yu-Jin, CAO Qian, LU Ling-Ling, SUN Xiao-Gong, GAO Hua, DIAO Sha-Sha, YANG Hui. Nrf3 Pathway was Involved in the Protection of Astrocyte Against the Neurotoxicity of Rotenone. China Biotechnology, 2010, 30(01): 19-24.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2010/V30/I01/19

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