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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。

Myoglobin Mediates Autophagy of NRK-52E in Rat Renal Tubular Epithelial Cells Via the Pink1/Parkin Signaling Pathway.

机构信息

Department of Emergency Medicine, General Hospital, Tianjin Medical University, Tianjin, China (mainland).

Institute of Disaster Medicine, Tianjin University, Tianjin, China (mainland).

出版信息

Med Sci Monit. 2020 Jul 22;26:e923045. doi: 10.12659/MSM.923045.

DOI:10.12659/MSM.923045
PMID:32697768
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7391798/
Abstract

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.

摘要

背景

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。

材料和方法

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。

结果

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。

结论

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。

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