山东医药2017，Vol.57Issue(15)：21-25,5.DOI:10.3969/j.issn.1002-266X.2017.15.006
帕金森病细胞模型的构建及氨基酸表达观察
Construction of Parkinson 's disease cell model and observation of amino acid expression
摘要
Abstract
Objective To establish the human neuroblastoma cell model with Parkinson's disease (PD), and screen out the amino acid biomarkers in PD cells. Methods Human neuroblastoma SH-SY5Y cells were divided into the normal group, solvent group and rotenone group. Cells in the rotenone group were given 0.15, 0.25 μmol/L rotenone, cells in the solvent group were given dimethyl sulfoxide (DMSO), and the normal group was not added with any drug. Cell viability was assessed by MTT assay at 24, 48 h after medication, and inverted microscope was used to observe the cell morphology. Then we determined the concentration (0.15 μmol/L) and treatment time (24 h) of rotenone which had the little effect according to the changes of cell viability and cell morphology. The expression of α-synuclein was detected by Western blot to validate the success of PD model. And 12 kinds of amino acids, including glutamic acid (Glu), serine (Ser), glutamine (Gln), histidine (His), glycine (Gly), alanine (Ala), proline (Pro), tryptophan (Trp ), phenylalanine (Phe), isoleucine (Ile), leucine (Leu), and lysine (Lys), in PD cells were determined by high performance liquid chromatography, and we analyzed the significant changed amino acids between solvent group and rotenone group with partial least squares discriminant analysis (PLS-DA). Amino acid biomarkers in the PD model were determined with a variable importance in the projection (VIP) value greater than 1 and a P value less than 0.05. Results The expression of α-synuclein in the cells of the rotenone group was significantly higher than that of the solvent group (P<0.05), indicating the successful construction of SH-SY5Y cell model with PD. At 24 h, the Glu, Ser, Gln, His, Gly, Pro, Phe, Ile, Leu, Lys levels in the rotenone group were lower than those in the solvent group (all P<0.05). At 48 h, the levels of 11 kinds of amino acid except Gln in the rotenone group were lower than those in the solvent group (all P<0.05). Glu, Ser, Gly, Pro were screened out as the amino acid biomarkers (At 24 h, VIP values were 1.016, 1.016, 1.016, 1.009; at 48 h, all VIP values were 1.001). Conclusion The PD cell model is successfully established by rotenone induction and Glu, Ser, Gly, and Pro may be the potential diagnostic biomarkers for PD.关键词
帕金森病/鱼藤酮/人神经母细胞瘤细胞/氨基酸Key words
Parkinson's disease/rotenone/human neuroblastoma cells/amino acid分类
医药卫生引用本文复制引用
李双,李双双,张晓莹,郭春燕,王治宝..帕金森病细胞模型的构建及氨基酸表达观察[J].山东医药,2017,57(15):21-25,5.基金项目
河北省教育厅重大项目(ZD2014075) （ZD2014075）
河北北方学院博士科研启动基金项目. （）
