Department of Emergency Medicine, General Hospital, Tianjin Medical University, Tianjin, China (mainland).
Institute of Disaster Medicine, Tianjin University, Tianjin, China (mainland).
Med Sci Monit. 2020 Jul 22;26:e923045. doi: 10.12659/MSM.923045.
BACKGROUND The aim of this study was to investigate whether myoglobin mediates the autophagy of NRK-52E via the Pink1/Parkin signaling pathway. MATERIAL AND METHODS Differentially-expressed genes were selected by PCR chip analysis of the autophagy signaling pathway. RT-PCR and Western blot analyses were used to detect the expressions of Pink1/Parkin and autophagy-related proteins in myoglobin-treated NRK-52E. LC3 double-labeled lentivirus was used to infect NRK-52E for observing autophagy. The role of myoglobin mediates autophagy was evaluated through Pink1-siRNA inhibition of the Pink1/Parkin signaling pathway. RESULTS Myoglobin acted on NRK-52E, caused differential expressions of Pink1, Parkin, and Beclin 1, increased apoptosis, and decreased cell viability. myoglobin increased the levels of Pink1, Beclin 1 and ATG5, decreased the levels of P62 and Parkin. The level of LC3II/LC3I showed significant elevation in NRK-52E cells at after incubated with 100 μmol/L myoglobin. Inhibiting Pink1/Parkin signaling pathway through Pink1-siRNA could alleviate myoglobin induced apoptosis, decrease the levels of Pink, Beclin1, ATG5, LC3II/LC3I, and elevate the levels of Parkin and P62. Moreover, the autophagy spots were reduced after silencing Pink1 in myoglobin-treated NRK-52E. CONCLUSIONS Myoglobin mediates the autophagy of NRK-52E in rat renal tubular epithelial cells via the Pink1/Parkin signaling pathway.
本研究旨在探讨肌红蛋白是否通过 Pink1/Parkin 信号通路介导 NRK-52E 的自噬。
通过自噬信号通路的 PCR 芯片分析选择差异表达基因。使用 RT-PCR 和 Western blot 分析检测肌红蛋白处理的 NRK-52E 中 Pink1/Parkin 和自噬相关蛋白的表达。使用 LC3 双标记慢病毒感染 NRK-52E 观察自噬。通过 Pink1-siRNA 抑制 Pink1/Parkin 信号通路评估肌红蛋白介导自噬的作用。
肌红蛋白作用于 NRK-52E,引起 Pink1、Parkin 和 Beclin 1 的差异表达,增加细胞凋亡,降低细胞活力。肌红蛋白增加 Pink1、Beclin 1 和 ATG5 的水平,降低 P62 和 Parkin 的水平。LC3II/LC3I 的水平在 NRK-52E 细胞孵育 100μmol/L 肌红蛋白后显著升高。通过 Pink1-siRNA 抑制 Pink1/Parkin 信号通路可减轻肌红蛋白诱导的细胞凋亡,降低 Pink1、Beclin1、ATG5、LC3II/LC3I 的水平,升高 Parkin 和 P62 的水平。此外,沉默肌红蛋白处理的 NRK-52E 中的 Pink1 后,自噬斑点减少。
肌红蛋白通过 Pink1/Parkin 信号通路介导大鼠肾小管上皮细胞 NRK-52E 的自噬。