The objective of this study was to understand if RNF13 can affect Parkinson’s disease (PD) model mice by modulating the endoplasmic reticulum stress (ERS)-mediated IRE1α-TRAF2-ASK1-JNK pathway. C57BL/6 mice injected with MPTP to establish PD mice models were divided into Control, MPTP, MPTP + sh-RNF13, and MPTP + sh-NC groups. Rotarod, balance beam, and open-field tests were used to assess the behavioral changes of experimental mice. Immunofluorescence assay was used to determine TH-positive expression in substantia nigra, TUNEL staining to detect apoptosis, and Western blotting to measure the expression of IRE1α-TRAF2-ASK1-JNK pathway. Besides, SH-SY5Y cells treated with MPP⁺ were assigned into Control, MPP⁺, MPP⁺ + sh-RNF13, and MPP⁺ + sh-NC groups in vitro to detect cell viability, apoptosis and Ca²⁺ level. When compared with those Control mice, MPTP mice showed decreased retention time spent on rotarod performance and prolonged time on balance beam test, as well as evident reductions in floor plane (FP) movements, moving time, moving distance, and mean velocity in open-field test, which had an obvious increase of TUNEL-positive cells, significant decrease of TH-positive cells, and remarkable up-regulations of RNF13, p-IRE1α/IRE1α, TRAF2, ASK1, and p-JNK/JNK. Meanwhile, MPTP mice treated with sh-RNF13 were improved in all above indexes. In vitro, MPP⁺ treated SH-SY5Y cells had decreased cell viability and increased cell apoptosis, as well as the upregulated IRE1α-TRAF2-ASK1-JNK pathway proteins and Ca²⁺ level. RNF13 knockdown improved all above indexes in SH-SY5Y cells treated with MPP⁺. Silencing RNF13 can alleviate motor dysfunction and dopamine neuronal damage in PD mice by inhibiting ERS-mediated IRE1α-TRAF2-ASK1-JNK pathway.

本研究的目的是了解 RNF13 是否可以通过调节内质网应激 (ERS) 介导的 IRE1α-TRAF2-ASK1-JNK 通路来影响帕金森病 (PD) 模型小鼠。注射MPTP建立PD小鼠模型的C57BL/6小鼠分为Control组、MPTP组、MPTP+sh-RNF13组和MPTP+sh-NC组。使用旋转棒、平衡木和开放场测试来评估实验小鼠的行为变化。免疫荧光法检测黑质中TH阳性表达，TUNEL染色检测细胞凋亡，Western blotting检测IRE1α-TRAF2-ASK1-JNK通路的表达。此外，用 MPP ⁺处理的 SH-SY5Y 细胞被分配到 Control、MPP ⁺、MPP ⁺ + sh-RNF13 和 MPP ⁺+ sh-NC 组体外检测细胞活力、细胞凋亡和 Ca ²⁺水平。与那些对照小鼠相比，MPTP 小鼠在旋转杆性能上花费的保留时间减少，平衡木测试时间延长，地板平面 (FP) 运动、移动时间、移动距离和开放式平均速度明显减少。田间试验，TUNEL阳性细胞明显增多，TH阳性细胞明显减少，RNF13、p-IRE1α/IRE1α、TRAF2、ASK1、p-JNK/JNK显着上调。同时，sh-RNF13处理的MPTP小鼠上述各项指标均有改善。在体外，MPP ⁺处理的 SH-SY5Y 细胞具有降低的细胞活力和增加的细胞凋亡，以及上调的 IRE1α-TRAF2-ASK1-JNK 通路蛋白和 Ca²⁺级。RNF13 敲低改善了用 MPP ⁺处理的 SH-SY5Y 细胞的所有上述指标。沉默 RNF13 可以通过抑制 ERS ​​介导的 IRE1α-TRAF2-ASK1-JNK 通路减轻 PD 小鼠的运动功能障碍和多巴胺神经元损伤。