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

  • Nephrology - Original Paper
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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.

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Funding

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

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Authors and Affiliations

  1. Department of Nephrology, Shanghai Ninth People’s Hospital, School of Medicine, Shanghai Jiao Tong University, 639 Zhizaoju Rd., Shanghai, 200011, China

    Yue Shen, Lulu Zhang & Feng Ding

  2. Department of Nephrology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, 160 Pujian Rd., Shanghai, 200127, China

    Yucheng Yan, Yingying Qian, Xuejing Guan & Leyi Gu

  3. Shanghai Institute of Immunology, School of Medicine, Shanghai Jiao Tong University, 227 South Chongqing Rd., Shanghai, 200025, China

    Liming Lu & Yuanyuan Qi

Authors
  1. Yue Shen
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  2. Yucheng Yan
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  3. Liming Lu
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  4. Yingying Qian
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  5. Xuejing Guan
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  6. Lulu Zhang
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  7. Yuanyuan Qi
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  8. Leyi Gu
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  9. Feng Ding
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Corresponding author

Correspondence to Yucheng Yan.

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Conflict of interest

The authors declare that they have no conflict of interest.

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

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Cite this article

Shen, Y., Yan, Y., Lu, L. et al. Klotho ameliorates hydrogen peroxide-induced oxidative injury in TCMK-1 cells. Int Urol Nephrol 50, 787–798 (2018). https://doi.org/10.1007/s11255-017-1765-x

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  • DOI: https://doi.org/10.1007/s11255-017-1765-x

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