Open Access

Effect of ATM on inflammatory response and autophagy in renal tubular epithelial cells in LPS‑induced septic AKI

  • Authors:
    • Chenfei Zheng
    • Ying Zhou
    • Yueyue Huang
    • Bicheng Chen
    • Minmin Wu
    • Yue Xie
    • Xinxin Chen
    • Mei Sun
    • Yi Liu
    • Chaosheng Chen
    • Jingye Pan
  • View Affiliations

  • Published online on: October 21, 2019     https://doi.org/10.3892/etm.2019.8115
  • Pages: 4707-4717
  • Copyright: © Zheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to explore the role of ataxia‑telangiectasia mutated (ATM) in lipopolysaccharide (LPS)‑induced in vitro model of septic acute kidney injury (AKI) and the association between ATM, tubular epithelial inflammatory response and autophagy. The renal tubular epithelial cell HK‑2 cell line was cultured and passaged, with HK‑2 cell injury induced by LPS. The effects of LPS on HK‑2 cell morphology, viability, ATM expression and inflammation were observed. Lentiviral vectors encoding ATM shRNA were constructed to knock down ATM expression in HK‑2 cells. The efficiency of ATM knockdown in HK‑2 cells was detected by western blot analysis and reverse transcription‑quantitative PCR (RT‑qPCR). HK‑2 cells transfected with the ATM shRNA lentivirus were used for subsequent experiments. Following ATM knockdown, corresponding controls were set up, and the effects of ATM on inflammation and autophagy were detected in HK‑2 cells using RT‑qPCR, western blotting and ELISA. After LPS stimulation, the HK‑2 cells were rounded into a slender or fusiform shape with poorly defined outlines. LPS treatment reduced cell viability in a partly dose‑dependent manner. LPS increased the expression of tumor necrosis factor‑α, interleukin (IL)‑1β and IL‑6, with the levels reaching its highest value at 10 µg/ml. IL‑6 and IL‑1β expression increased with increasing LPS concentration. These findings suggest that LPS reduced HK‑2 cell viability whilst increasing the expression of inflammatory factors. Following transfection with ATM shRNA, expression levels of key autophagy indicators microtubule associated protein 1 light chain 3α I/II ratio and beclin‑1 in the two ATM shRNA groups were also significantly reduced compared with the NC shRNA group. In summary, downregulation of ATM expression in HK‑2 cells reduced LPS‑induced inflammation and autophagy in sepsis‑induced AKI in vitro, suggesting that LPS may induce autophagy in HK‑2 cells through the ATM pathway leading to the upregulation of inflammatory factors.
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December 2019
Volume 18 Issue 6

Print ISSN: 1792-0981
Online ISSN:1792-1015

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APA
Zheng, C., Zhou, Y., Huang, Y., Chen, B., Wu, M., Xie, Y. ... Pan, J. (2019). Effect of ATM on inflammatory response and autophagy in renal tubular epithelial cells in LPS‑induced septic AKI. Experimental and Therapeutic Medicine, 18, 4707-4717. https://doi.org/10.3892/etm.2019.8115
MLA
Zheng, C., Zhou, Y., Huang, Y., Chen, B., Wu, M., Xie, Y., Chen, X., Sun, M., Liu, Y., Chen, C., Pan, J."Effect of ATM on inflammatory response and autophagy in renal tubular epithelial cells in LPS‑induced septic AKI". Experimental and Therapeutic Medicine 18.6 (2019): 4707-4717.
Chicago
Zheng, C., Zhou, Y., Huang, Y., Chen, B., Wu, M., Xie, Y., Chen, X., Sun, M., Liu, Y., Chen, C., Pan, J."Effect of ATM on inflammatory response and autophagy in renal tubular epithelial cells in LPS‑induced septic AKI". Experimental and Therapeutic Medicine 18, no. 6 (2019): 4707-4717. https://doi.org/10.3892/etm.2019.8115