Key Laboratory for Biotechnology on Medicinal Plants of Jiangsu Province, School of Life Science, Jiangsu Normal University, Xuzhou, China.
College of Health Sciences, Jiangsu Normal University, Xuzhou, China.
J Cell Physiol. 2019 Jun;234(6):9033-9044. doi: 10.1002/jcp.27577. Epub 2018 Oct 26.
Development of effective therapeutic drugs for Parkinson's disease (PD) is of great importance. Aberrant microRNA (miRNA) expression has been identified in postmortem human PD brain samples, in vitro and in vivo PD models. However, the role of miR-342-3p in PD has been understudied. The study explores the effects of miR-342-3p on expression of glutamate (Glu) transporter, and dopaminergic neuron apoptosis and proliferation by targeting p21-activated kinase 1 (PAK1) through the Wnt signaling pathway in PD mice. After establishment of PD mouse models, gain- or loss-of-function assay was performed to explore the functional role of miR-342-3p in PD. Number of apoptotic neurons and Glu concentration was then determined. Subsequently, PC12 cells were treated with miR-342-3p mimic, miR-342-3p inhibitor, dickkopf-1 (DKK1), and miR-342-3p inhibitor + DKK1. The expression of miR-342-3p, PAK1, the Wnt signaling pathway-related and apoptosis-related genes, Glutamate transporter subtype 1 (GLT-1), l-glutamate/ l-aspartate transporter (GLAST), tyrosine hydroxylase (TH) was measured. Also, cell viability and apoptosis were evaluated. PD mice exhibited increased miR-342-3p, while decreased expression of PAK1, GLT-1, GLAST, TH, and the Wnt signaling pathway-related and antiapoptosis genes. miR-342-3p downregulation could promote expression of PAK1, the Wnt signaling pathway-related and antiapoptosis genes. GLT-1, GLAST, and TH as well as cell viability, but reduce cell apoptosis rate. The results indicated that suppression of miR-342-3p improves expression of Glu transporter and promotes dopaminergic neuron proliferation while suppressing apoptosis through the Wnt signaling pathway by targeting PAK1 in mice with PD.
开发有效的帕金森病（PD）治疗药物非常重要。已经在尸检的人类 PD 脑样本、体外和体内 PD 模型中发现异常的 microRNA（miRNA）表达。然而，miR-342-3p 在 PD 中的作用尚未得到充分研究。本研究通过靶向 PD 小鼠中的 P21 激活激酶 1（PAK1），通过 Wnt 信号通路探索 miR-342-3p 对谷氨酸（Glu）转运体表达以及多巴胺能神经元凋亡和增殖的影响。建立 PD 小鼠模型后，进行增益或功能丧失测定以探索 miR-342-3p 在 PD 中的功能作用。然后测定凋亡神经元的数量和 Glu 浓度。随后，用 miR-342-3p 模拟物、miR-342-3p 抑制剂、Dickkopf-1（DKK1）和 miR-342-3p 抑制剂+DKK1 处理 PC12 细胞。测量 miR-342-3p、PAK1、Wnt 信号通路相关和凋亡相关基因、谷氨酸转运体亚型 1（GLT-1）、l-谷氨酸/l-天冬氨酸转运体（GLAST）、酪氨酸羟化酶（TH）的表达。还评估了细胞活力和凋亡。PD 小鼠表现出 miR-342-3p 增加，而 PAK1、GLT-1、GLAST、TH 和 Wnt 信号通路相关和抗凋亡基因表达降低。miR-342-3p 的下调可以促进 PAK1、Wnt 信号通路相关和抗凋亡基因的表达。GLT-1、GLAST 和 TH 以及细胞活力，但降低细胞凋亡率。结果表明，通过靶向 PAK1 抑制 miR-342-3p 可改善 PD 小鼠中 Glu 转运体的表达，并通过 Wnt 信号通路促进多巴胺能神经元增殖，同时抑制细胞凋亡。
