Establishment of a cellular model of Parkinson disease by treating differentiated SH-SY5Y cells with α-synuclein preformed fibrils
10.12007/j.issn.0258-4646.2025.01.002
- VernacularTitle:利用α-突触核蛋白预制纤维作用于分化的SH-SY5Y细胞建立帕金森病细胞模型
- Author:
Jingxi SU
1
;
Qiong SONG
;
Zihan JING
;
Lianghao CHEN
;
Chunlin ZOU
Author Information
1. 广西医科大学基础医学院转化医学研究中心长寿与老年相关疾病教育部重点实验室,神经科学研究所广西脑科学研究重点实验室,南宁 530021
- Publication Type:Journal Article
- Keywords:
Parkinson disease;
α-synuclein preformed fibrils;
retinoic acid;
SH-SY5Y cells;
cellular model
- From:
Journal of China Medical University
2025;54(1):5-11
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a cellular model of Parkinson disease by treating retinoic acid(RA)-differentiated SH-SY5Y cells with α-synuclein preformed fibrils(α-Syn PFF).Methods SH-SY5Y cells were divided into undifferentiated and RA-differentiated groups.The expression levels of tyrosine hydroxylase(TH),dopamine transporter(DAT),lymphocyte activation gene 3(LAG3),and Nestin proteins in the cells were detected using Western blotting,whereas those of microtubule-associated protein 2(MAP2)and neu-ronal nuclei(NeuN)were detected using immunofluorescence staining.Furthermore,RA-treated SH-SY5Y cells were divided into control and α-Syn PFF groups,and their levels of chromatin condensation were detected using Hoechst 33342 staining.Nitric oxide(NO)levels were measured using a NO assay kit.Additionally,the protein levels of TH,poly(ADP-ribose)(PAR),and poly(ADP-ribose)poly-merase(PARP)in these cells were detected using Western blotting,whereas their expression levels of phosphorylated α-Syn(pS129-α-Syn)and phosphorylated histone H2AX(γH2AX)were detected using immunofluorescence staining.Results Treatment with RA resulted in a reduction in cell body size and the elongation of protrusions in SH-SY5Y cells.The results of Western blotting showed that RA treatment could increase the TH,DAT,and LAG3 levels and decrease the Nestin level in SH-SY5Y cells(P＜0.05).α-Syn PFF treat-ment decreased the TH protein level and increased the PAR,PARP-l,and cleaved PARP-1 levels in differentiated SH-SY5Y cells(P＜0.05).According to the immunofluorescence results,RA treatment increased the expression levels of MAP2 and NeuN in SH-SY5Y cells(P＜0.001).The α-Syn PFF treatment increased the expression levels of γH2AX and pS129-α-Syn in RA-differentiated SH-SY5Y cells(P＜0.01).The Hoechst 33342 staining results showed that α-Syn PFF treatment led to chromatin condensation in the differentiated SH-SY5Y cells(P＜0.001)and increased the NO levels(P＜0.01).Conclusion A cellular model of Parkinson disease can be established by treating RA-differentiated SH-SY5Y cells with α-Syn PFF.
