Sang Qiuling, Liu Xiaoyang, Wang Libo, Qi Ling, Sun Wenping, Wang Weiyao, Sun Yajuan, Zhang Haina
Department of Neurology, China-Japan Union Hospital of Jilin University, Changchun, Jilin 130033, China.
Department of Pathophysiology, Jilin Medical University, Jilin, Jilin 132013, China.
Aging (Albany NY). 2018 Jun 28;10(6):1281-1293. doi: 10.18632/aging.101466.
We aimed to explore the mechanism of pramipexole (PPX) actions in the treatment of Parkinson's disease (PD). Genes related to PD and PPX were screened through bioinformatics retrieval. The PD model was constructed by applying 1-methyl-4-phenylpyridinium (MMP+). The RNA expression levels of circSNCA, , apoptosis-related genes (, , , and ) and miR-7 were detected by qRT-PCR. Protein expression was determined by western blot. The interactions between circSNCA-miR-7- were verified by dual luciferase assay and immunofluorescence localization. Cell viability was determined by MTT assay. and circSNCA expression levels in PD were downregulated after PPX treatment, consistent with the levels of pro-apoptotic genes. CircSNCA increased expression by downregulating miR-7 in PD as a competitive endogenous RNA (ceRNA). Lower circSNCA expression was associated with the reduced expression of pro-apoptotic (, , and ) proteins. CircSNCA downregulation could decrease apoptosis and induce autophagy in PD. In conclusion, the downregulation of circSNCA by PPX treatment reduced cell apoptosis and promoted cell autophagy in PD via a mechanism that served as a miR-7 sponge to upregulate .
我们旨在探究普拉克索(PPX)治疗帕金森病(PD)的作用机制。通过生物信息学检索筛选与PD和PPX相关的基因。应用1-甲基-4-苯基吡啶离子(MMP⁺)构建PD模型。采用qRT-PCR检测circSNCA、凋亡相关基因(、、、和)以及miR-7的RNA表达水平。通过蛋白质印迹法测定蛋白质表达。采用双荧光素酶报告基因检测和免疫荧光定位验证circSNCA与miR-7之间的相互作用。通过MTT法测定细胞活力。PPX治疗后,PD中circSNCA的表达水平下调,这与促凋亡基因的水平一致。在PD中,circSNCA作为竞争性内源性RNA(ceRNA)通过下调miR-7增加的表达。较低的circSNCA表达与促凋亡蛋白(、、和)表达降低有关。circSNCA下调可减少PD中的细胞凋亡并诱导自噬。总之,PPX治疗下调circSNCA通过作为miR-7海绵上调从而减少PD中的细胞凋亡并促进细胞自噬。