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International Urology and Nephrology

, Volume 50, Issue 4, pp 787–798 | Cite as

Klotho ameliorates hydrogen peroxide-induced oxidative injury in TCMK-1 cells

  • Yue Shen
  • Yucheng Yan
  • Liming Lu
  • Yingying Qian
  • Xuejing Guan
  • Lulu Zhang
  • Yuanyuan Qi
  • Leyi Gu
  • Feng Ding
Nephrology - Original Paper
  • 243 Downloads

Abstract

Purpose

Defects in Klotho gene expression in mice result in a vulnerability to oxidative injuries. We aimed to identify the expression of Klotho in a mouse tubular epithelial (TCMK-1) cell line, and also to investigate changes in Klotho expression induced by oxidative stress and the potential role of intra- and extracellular Klotho protein.

Methods

During exposure to hydrogen peroxide (H2O2), an overexpression of the Klotho gene was induced and exogenous Klotho protein was added in TCMK-1 cells. The generation of reactive oxidative species (ROS) was examined by flow cytometry, and cell viability was assessed by Cell Counting Kit-8. Cellular apoptosis was determined by flow cytometry and Hoechst 33258 staining followed by Western blotting to evaluate the expression of Klotho, antioxidant enzymes, and apoptosis-associated proteins.

Results

While H2O2 significantly suppressed Klotho expression, cell viability, and the expression of antioxidant enzymes in a concentration-dependent manner, cellular apoptosis was increased and p38/MAPK and JNK/MAPK were activated. Intra- and extracellular Klotho remarkably ameliorated viability inhibition, ROS generation, and cellular apoptosis induced by H2O2. Intra- and extracellular Klotho also reversed the loss of antioxidant enzymes, the elevation of cleaved Caspase-3 and Bax/Bcl-2, and the phosphorylation of JNK/MAPK and p38/MAPK.

Conclusions

Klotho has posed antioxidant and anti-apoptotic effects on oxidative injuries in TCMK-1 cells, which might be partially related to its inhibition of JNK/MAPK and p38/MAPK phosphorylation and subsequent elevation of antioxidant enzymes. Increasing Klotho expression has played a protective role against oxidative stress in tubular epithelial cells.

Keywords

Klotho Oxidative stress Apoptosis MAPK 

Notes

Funding

This study was sponsored by the National Natural Science Foundation of China (No. 81170687, 81470918), and Shanghai Ninth People’s Hospital (JYLJ007).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Informed consent

All subjects and control subjects gave a written informed consent to participate in the current study.

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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2017

Authors and Affiliations

  • Yue Shen
    • 1
  • Yucheng Yan
    • 2
  • Liming Lu
    • 3
  • Yingying Qian
    • 2
  • Xuejing Guan
    • 2
  • Lulu Zhang
    • 1
  • Yuanyuan Qi
    • 3
  • Leyi Gu
    • 2
  • Feng Ding
    • 1
  1. 1.Department of Nephrology, Shanghai Ninth People’s Hospital, School of MedicineShanghai Jiao Tong UniversityShanghaiChina
  2. 2.Department of Nephrology, Ren Ji Hospital, School of MedicineShanghai Jiao Tong UniversityShanghaiChina
  3. 3.Shanghai Institute of Immunology, School of MedicineShanghai Jiao Tong UniversityShanghaiChina

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