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（英文）

发布时间：2018-11-24 20:24

【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300μmol/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300μmol/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。
[Abstract]:In the early stage of Parkinson's disease (Parkinson's disease,PD), iron selectively accumulates in the substantia nigra compact zone, and the iron content in the remaining dopaminergic neurons increases, suggesting that the increase of iron may be a key factor in the occurrence of PD. Iron regulatory proteins (iron regulatory proteins,IRPs) IRP1 and IRP2 can bind to iron reactive elements in iron transport and storage proteins and play an important role in maintaining iron homeostasis in cells. FBXL5) participates in the regulation of iron metabolism through the degradation of IRP2 by ubiquitin proteasome pathway. Some studies have shown that nitric oxide (nitric oxide,NO) can enhance the activity of IRP1, but the effect on the expression of IRP2 is not clear. In this study, the donor of NO (sodium nitroprusside,SNP) was selected as the treatment drug to study the effect of sodium nitroprusside,SNP on the expression of FBXL5 and IRP2 protein in cultured SH-SY5Y cells. The results of cell viability test showed that SNP could damage SH-SY5Y cells in a dose-dependent manner and reduce cell viability. Flow cytometry showed that after 100 渭 mol/L and 300 渭 mol/L SNP treatment, the mitochondrial membrane potential of the cells decreased by 45% and 60, respectively. In addition, Western blotting results showed that 300 渭 mol/L SNP could increase the protein expression of FBXL5 and decrease the protein expression of IRP2 by 39% and 46% respectively. These results suggest that SNP can induce mitochondrial dysfunction in SH-SY5Y cells and up-regulate the expression of FBXL5 and down-regulate the expression of IRP2.
【作者单位】：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;
【基金】：supported by the National Natural Science Foundation of China(No.31471114,31500837 and 31540075) Taishan Scholarship,the Key Research and Development Program of Shandong Province,China(No.2016GSF201053) Qingdao Municipal Science and Technology Project(No.16-6-2-2-nsh) the Shandong Provincial Natural Science Foundation of China(No.BS2015SW022)
【分类号】：R742.5

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