1. Academic Validation
  2. Inhibition of PLK3 Attenuates Tubular Epithelial Cell Apoptosis after Renal Ischemia-Reperfusion Injury by Blocking the ATM/P53-Mediated DNA Damage Response

Inhibition of PLK3 Attenuates Tubular Epithelial Cell Apoptosis after Renal Ischemia-Reperfusion Injury by Blocking the ATM/P53-Mediated DNA Damage Response

  • Oxid Med Cell Longev. 2022 Jun 24;2022:4201287. doi: 10.1155/2022/4201287.
Weiming Deng 1 2 Xiangling Wei 1 Zhenwei Xie 1 Rui Zhang 1 Zhanwen Dong 1 Jinhua Zhang 1 You Luo 1 Qingdi Cheng 1 Ruojiao Wang 1 Heng Li 1 Ning Na 1
Affiliations

Affiliations

  • 1 Department of Kidney Transplantation, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510630, China.
  • 2 The First Affiliated Hospital, Department of Urology, Hengyang Medical School, University of South China, Hengyang, Hunan 421001, China.
Abstract

Objective: Renal ischemia-reperfusion (I/R) injury is a major cause of acute kidney injury (AKI) in transplanted kidneys. This study was aimed at exploring the role of PLK3 (polo-like kinase 3) in renal I/R injury, focusing on its relationship with oxidative stress-induced DNA damage and renal tubular epithelial cell (TEC) Apoptosis.

Methods: TRAP-seq data from the development dataset GSE52004 and the validation dataset GSE121191 were analyzed using GEO2R. PLK3 overexpression plasmids and targeted silencing siRNAs were used in a model of hypoxia/reoxygenation (H/R) injury, and rAAV-9-PLK3-KD were administered to C57BL/6J mice exposed to I/R injury. The ATM-specific inhibitor KU-60019 was used to block the DNA damage response (DDR). Western blotting was performed to measure DDR- and apoptosis-associated protein expression. Cell viability was measured by CCK-8 reagent, and Apoptosis was examined by flow cytometry and TUNEL assay. Furthermore, the fluorescent probes H2DCFH-DA and DHE were used to measure ROS production in vitro. The MDA level and SOD activity were measured to assess oxidative stress in vivo. KIM-1 staining and Scr and BUN were used to evaluate kidney injury.

Results: The mRNA and protein levels of PLK3 were markedly increased in the H/R injury and I/R injury models. GO terms showed that PLK3 was mainly involved in oxidative stress and DNA damage after renal I/R injury. Overexpression of PLK3 decreased cell viability and increased Apoptosis. In contrast, targeted silencing of PLK3 expression decreased the Bax/Bcl-2 ratio by decreasing P53 phosphorylation, thereby reducing TEC Apoptosis. Furthermore, KU-60019 reduced PLK3 activation and DDR-induced Apoptosis, while overexpression of PLK3 reversed the mitigating effect of KU-60019 on TEC Apoptosis. Similarly, rAAV-9-PLK3 KD mice exhibited a lower rate of TEC Apoptosis and milder renal damage after I/R injury.

Conclusion: We demonstrate for the first time that PLK3 is involved in oxidative stress-induced DNA damage and TEC Apoptosis in renal I/R injury. Inhibition of PLK3 attenuates TEC Apoptosis after I/R injury by blocking the ATM/P53-mediated DDR. Therefore, PLK3 may serve as a potential therapeutic target for ischemic AKI.

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  • HY-12061
    99.43%, ATM/ATR抑制剂