Department of Neurology, the Affiliated Hospital of Xuzhou Medical University, Xuzhou, China.
Eur Rev Med Pharmacol Sci. 2020 Nov;24(21):11192-11198. doi: 10.26355/eurrev_202011_23607.
The aim of this study was to explore the effect of micro ribonucleic acid (miR)-133b on 1-methyl-4-phenylpyridinium ion (MPP+)-induced apoptosis in the Parkinson's disease (PD) model.
PC12 cells were induced by different concentrations of MPP+ to establish the PD cell model. Subsequently, the survival rate of PC12 cells was detected using Cell Counting Kit-8 (CCK-8) assay. Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) was used to detect the expression of miR-133b in the PD model induced by different concentrations of MPP+. Next, PC12 cells were transfected with miR-133b mimic and miR-negative control (NC), and divided into MPP+ group, MPP+ + miR-NC group and MPP+ + miR-133b mimic group. Transfection efficiency was verified using qRT-PCR. The apoptosis of cells was detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Moreover, the expressions of extracellular signal-regulated kinase 1/2 (ERK1/2) and phosphorylated (p)-ERK1/2 were determined using Western blotting.
After MPP+ treatment, the survival rate of PC12 cells significantly declined (p<0.05). MPP+ exhibited toxicity against PC12 cells in a concentration-dependent manner. Meanwhile, cell survival rate decreased remarkably with the increase of MPP+ concentration (p<0.05). With increased concentration of MPP+, the expression of miR-133b in the PD cell model declined significantly (p<0.05). The apoptosis of PC12 cells was remarkably inhibited by overexpression of miR-133b in the PD cell model (p<0.05). In addition, the protein expression of p-ERK1/2 in PC12 cells was notably reduced after overexpression of miR-133b in the PD cell model (p<0.05).
MiR-133b is lowly expressed in the PD cell model. Furthermore, overexpression of miR-133b inhibits cell apoptosis in the PD cell model by regulating the ERK1/2 signaling pathway.
本研究旨在探讨微小 RNA-133b（miR-133b）对 1-甲基-4-苯基吡啶离子（MPP+）诱导的帕金森病（PD）模型细胞凋亡的影响。
用不同浓度的 MPP+诱导 PC12 细胞，建立 PD 细胞模型。然后，用细胞计数试剂盒-8（CCK-8）法检测 PC12 细胞的存活率。采用实时定量逆转录聚合酶链反应（qRT-PCR）检测不同浓度 MPP+诱导的 PD 模型中 miR-133b 的表达。接下来，用 miR-133b 模拟物和 miR-阴性对照（NC）转染 PC12 细胞，并分为 MPP+组、MPP+ + miR-NC 组和 MPP+ + miR-133b 模拟物组。用 qRT-PCR 验证转染效率。用末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记（TUNEL）法检测细胞凋亡。此外，用 Western blot 法检测细胞外信号调节激酶 1/2（ERK1/2）和磷酸化（p）-ERK1/2 的表达。
MPP+处理后，PC12 细胞的存活率明显下降（p<0.05）。MPP+对 PC12 细胞具有浓度依赖性毒性。同时，随着 MPP+浓度的增加，细胞存活率显著下降（p<0.05）。随着 PD 细胞模型中 MPP+浓度的增加，miR-133b 的表达明显下降（p<0.05）。PD 细胞模型中 miR-133b 的过表达显著抑制了 PC12 细胞的凋亡（p<0.05）。此外，PD 细胞模型中 miR-133b 的过表达显著降低了 PC12 细胞中 p-ERK1/2 的蛋白表达（p<0.05）。
miR-133b 在 PD 细胞模型中低表达。此外，miR-133b 的过表达通过调节 ERK1/2 信号通路抑制 PD 细胞模型中的细胞凋亡。
