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硝普钠对SH-SY5Y细胞中F-box富含亮氨酸重复蛋白5和铁调节蛋白2表达的调节作用

魏杰, 李勇, 焦倩, 杜希恂, 姜宏^*

青岛大学医学院生理学教研室，山东省神经相关疾病重点实验室，山东省神经退变疾病协同创新中心，青岛 266071

摘要

帕金森病(Parkinson’s disease, PD)患者早期铁选择性聚集在黑质致密带，残存的多巴胺能神经元内铁含量增高，说明铁升高可能是PD发生的一个关键因素。铁调节蛋白(iron regulatory proteins, IRPs) IRP1和IRP2可与铁转运和储存蛋白中的铁反应元件相结合，对维持细胞铁稳态起重要作用。F-box富含亮氨酸重复蛋白5 (F-box and leucine-rich repeat protein 5, FBXL5)通过泛素蛋白酶体途径降解IRP2而参与铁代谢的调控。有研究显示一氧化氮(nitric oxide, NO)能够增强IRP1的活性，但对IRP2的表达影响尚未明确。本研究选择NO的供体硝普纳(sodium nitroprusside, SNP)作为处理药物，研究其对体外培养的SH-SY5Y细胞内FBXL5和IRP2蛋白表达的影响。细胞活力检测结果显示SNP可剂量依赖性地损伤SH-SY5Y细胞，降低细胞存活率。流式细胞术结果显示，经过100和300 μmol/L的SNP处理后，细胞线粒体膜电位分别降低45%和60%。此外，Western blotting结果显示300 μmol/L SNP能够引起细胞内FBXL5的蛋白表达量升高39%，而使IRP2的蛋白表达量降低46%。以上结果提示，SNP可造成SH-SY5Y细胞的线粒体功能障碍，并上调FBXL5的表达和下调IRP2的表达。

关键词： 帕金森病; 铁 ; 硝普纳 ; F-box富含亮氨酸重复蛋白5 ; 铁调节蛋白2

分类号：Q42;R338

Influence of sodium nitroprusside on expressions of FBXL5 and IRP2 in SH-SY5Y cells

WEI Jie, LI Yong, JIAO Qian, DU Xi-Xun, JIANG Hong^*

Department of Physiology, Shandong Provincial Key Laboratory of Pathogenesis and Prevention of Neurological Disorders, Shandong Provincial Collaborative Innovation Center for Neurodegenerative Disorders, Medical College of Qingdao University, Qingdao 266071, China

Abstract

Iron accumulation in the brain is associated with the pathogenesis of Parkinson’s disease (PD). Misexpression of some iron transport and storage proteins is related to iron dyshomeostasis. Iron regulatory proteins (IRPs) including IRP1 and IRP2 are cytosolic proteins that play important roles in maintaining cellular iron homeostasis. F-box and leucine-rich repeat protein 5 (FBXL5) is involved in the regulation of iron metabolism by degrading IRP2 through the ubiquitin-proteasome system. Nitric oxide (NO) enhances the binding activity of IRP1, but its effect on IRP2 is ambiguous. Therefore, in the present study, we aim to determine whether sodium nitroprusside (SNP), a NO donor, regulates FBXL5 and IRP2 expression in cultured SH-SY5Y cells. MTT assay revealed that treatment of SNP attenuated the cell viability in a dose-dependent manner. Flow cytometry test showed that 100 and 300 μmol/L SNP administration significantly reduced the mitochondrial membrane potential by 45% and 60%, respectively. Moreover, Western blotting analysis demonstrated that 300 μmol/L SNP significantly increased FBXL5 expression by about 39%, whereas the expression of IRP2 was decreased by 46%, correspondingly. These findings provide evidence that SNP could induce mitochondrial dysfunction, enhance FBXL5 expression and decrease IRP2 expression in SH-SY5Y cells.

Key words: Parkinson’s disease; iron ; sodium nitroprusside ; FBXL5 ; IRP2

收稿日期：2016-12-20　　录用日期：2017-04-05

通讯作者：姜宏　　E-mail: hongjiang@qdu.edu.cn

引用本文：

魏杰, 李勇, 焦倩, 杜希恂, 姜宏. 硝普钠对SH-SY5Y细胞中F-box富含亮氨酸重复蛋白5和铁调节蛋白2表达的调节作用[J]. 生理学报 2017; 69 (3): 261-266.

WEI Jie, LI Yong, JIAO Qian, DU Xi-Xun, JIANG Hong. Influence of sodium nitroprusside on expressions of FBXL5 and IRP2 in SH-SY5Y cells. Acta Physiol Sin 2017; 69 (3): 261-266