Department of Anatomy, Faculty of Science, Mahidol University, Bangkok, Thailand.
Chakri Naruebodindra Medical Institute, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Samut Prakan, Thailand.
ScientificWorldJournal. 2023 Apr 3;2023:1335201. doi: 10.1155/2023/1335201. eCollection 2023.
The tissue-specific protein eEF1A2 has been linked to the development of neurological disorders. The role of eEF1A2 in the pathogenesis of Parkinson's disease (PD) has yet to be investigated. The aim of this study was to determine the potential neuroprotective effects of eEF1A2 in an MPP model of PD. Differentiated SH-SY5Y cells were transfected with eEF1A2 siRNA, followed by MPP exposure. The expression of p-Akt1 and p-mTORC1 was determined using Western blotting. The expression of p53, Bax, Bcl-2, and caspase-3 was evaluated using qRT-PCR. Cleaved caspase-3 levels and Annexin V/propidium iodide flow cytometry were used to determine apoptosis. The effects of PI3K inhibition were examined. The results showed that eEF1A2 siRNA significantly reduced the eEF1A2 expression induced by MPP. MPP treatment activated Akt1 and mTORC1; however, eEF1A2 knockdown suppressed this activation. In eEF1A2-knockdown cells, MPP treatment increased the expression of p53 and caspase-3 mRNA levels as well as increased apoptotic cell death when compared to MPP treatment alone. In cells exposed to MPP, upstream inhibition of the Akt/mTOR pathway, by either LY294002 or wortmannin, inhibited the phosphorylation of Akt1 and mTORC1. Both PI3K inhibitors increased eEF1A2 expression in cells, whether or not they were also treated with MPP. In conclusion, eEF1A2 may function as a neuroprotective factor against MPP, in part by regulating the Akt/mTOR pathway upstream.
组织特异性蛋白 eEF1A2 与神经发育障碍有关。eEF1A2 在帕金森病 (PD) 发病机制中的作用尚未被研究。本研究旨在确定 eEF1A2 在 MPP 诱导的 PD 模型中的潜在神经保护作用。用 eEF1A2 siRNA 转染分化的 SH-SY5Y 细胞，然后用 MPP 暴露。用 Western blot 测定 p-Akt1 和 p-mTORC1 的表达。用 qRT-PCR 评估 p53、Bax、Bcl-2 和 caspase-3 的表达。用 cleaved caspase-3 水平和 Annexin V/碘化丙啶流式细胞术测定细胞凋亡。检查了 PI3K 抑制的效果。结果表明，eEF1A2 siRNA 显著降低了 MPP 诱导的 eEF1A2 表达。MPP 处理激活了 Akt1 和 mTORC1；然而，eEF1A2 敲低抑制了这种激活。在 eEF1A2 敲低细胞中，与单独 MPP 处理相比，MPP 处理增加了 p53 和 caspase-3 mRNA 水平的表达，并增加了凋亡细胞死亡。在暴露于 MPP 的细胞中，Akt/mTOR 通路的上游抑制，无论是通过 LY294002 还是wortmannin，都抑制了 Akt1 和 mTORC1 的磷酸化。两种 PI3K 抑制剂均增加了细胞中的 eEF1A2 表达，无论它们是否也用 MPP 处理。总之，eEF1A2 可能作为一种神经保护因子发挥作用，部分通过调节 Akt/mTOR 通路的上游。
