Department of Ophthalmology, The First Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
Department of Neurology, The First Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
Curr Neurovasc Res. 2021;18(1):102-112. doi: 10.2174/1567202618666210531141833.
Parkinson's disease (PD) is a neurodegenerative disorder caused by the progressive loss of dopaminergic neurons. Canopy fibroblast growth factor signaling regulator 2 (CNPY2) is down-regulated in this disease, but its functions are unknown.
This study investigates the effects and regulation of CNPY2 in the apoptosis of neurons in PD.
We established a PD model in vivo by a five consecutive days-injection of 1-methyl-4- phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) to mice. In vitro, the human SH-SY5Y neuroblastoma cells, after differentiation, were treated with 1-Methyl-4-phenylpyridinium iodide (MPP+) for modeling. The cells were transfected with a recombinant vector overexpressing CNPY2 followed by MPP+ treatment. Expression of CNPY2 and proteins related to apoptosis was detected by real-time PCR, western blot, or immunofluorescence staining. The ROS level and mitochondrial membrane potential were determined by flow cytometry. Cell viability and apoptosis were measured by MTT assay and TUNEL staining.
CNPY2 level was down-regulated both in the brain and retina of PD mice and also inhibited in neurons by MPP+ in vitro. Overexpression of CNPY2 repressed the level of Bax and cleaved caspase-3, enhanced Bcl-2 level, and promoted neurite length under MPP+ treatment. CNPY2 overexpression reduced the accumulation of ROS and mitochondria dysfunction in neurons. The AKT/ GSK3β signaling pathway was activated by overexpressed CNPY2 to inhibit MPP+-induced neuronal apoptosis, which was confirmed using an AKT inhibitor MK-2206 2HCl.
CNPY2 alleviates oxidative stress, mitochondria dysfunction, and apoptosis of neurons induced by MPP+ by activating the AKT/ GSK3β signaling pathway.
帕金森病（PD）是一种由多巴胺能神经元进行性丧失引起的神经退行性疾病。在这种疾病中，穹窿成纤维细胞生长因子信号调节因子 2（CNPY2）下调，但它的功能未知。
本研究旨在探讨 CNPY2 在 PD 神经元凋亡中的作用及其调控机制。
我们通过连续 5 天注射 1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）建立 PD 模型，在体研究。体外，人 SH-SY5Y 神经母细胞瘤细胞分化后用 1-甲基-4-苯基吡啶碘盐（MPP+）处理构建模型。转染过表达 CNPY2 的重组载体后用 MPP+处理，通过实时 PCR、western blot 或免疫荧光染色检测 CNPY2 和凋亡相关蛋白的表达。通过流式细胞术检测 ROS 水平和线粒体膜电位。通过 MTT 法和 TUNEL 染色检测细胞活力和凋亡。
PD 小鼠的大脑和视网膜中 CNPY2 水平下调，体外 MPP+也抑制神经元中 CNPY2 的表达。过表达 CNPY2 抑制 Bax 和 cleaved caspase-3 的水平，增加 Bcl-2 的水平，并促进 MPP+处理后的神经元突起长度。CNPY2 过表达减少神经元中 ROS 的积累和线粒体功能障碍。过表达的 CNPY2 通过激活 AKT/GSK3β 信号通路抑制 MPP+诱导的神经元凋亡，这一过程可被 AKT 抑制剂 MK-2206 2HCl 所证实。
CNPY2 通过激活 AKT/GSK3β 信号通路缓解 MPP+诱导的神经元氧化应激、线粒体功能障碍和凋亡。
