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NF-κB-dependent increase in tissue factor expression is responsible for hypoxic podocyte injury

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Abstract

Background

Fibrin deposition within glomeruli is commonly seen in kidney biopsy specimens, suggesting enhanced coagulant activity. Tissue factor (TF) is a coagulation factor which is also related to various biological effects, and TF is upregulated by hypoxia in cancer cells. Recently, hypoxic podocyte injury has been proposed, therefore, we investigated TF expression in hypoxia.

Methods

Conditionally immortalized human podocytes were differentiated and treated under hypoxic or normoxic conditions. mRNA expressions of TF and tissue factor pathway inhibitor (TFPI) were analyzed by quantitative RT-PCR. Protein levels of TF and TFPI were tested by enzyme-linked immunosorbent assay. We employed small interfering RNA (siRNA) to temporary knockdown early growth response protein 1 (Egr-1), hypoxia-inducible factor-1α (HIF-1α) and TF. The expression of CD2-associated protein (CD2AP) mRNA and phalloidin staining was examined to assess podocyte injury.

Results

Hypoxia increased mRNA expression of TF (6 h: 2.3 ± 0.05 fold, p < 0.001, 24 h: 5.6 ± 2.4 fold, p < 0.05) and suppressed TFPI (6 h: 0.54 ± 0.04 fold, p < 0.05, 24 h: 0.24 ± 0.06 fold, p < 0.001) compared with normoxia. Similarly, protein levels of TF were increased and TFPI were decreased. Egr-1 siRNA did not change TF mRNA expression. Pyrrolidine dithiocarbamate (PDTC), a nuclear factor kappa B (NF-κB) inhibitor, significantly reduced hypoxia induced TF expression, and HIF-1α knockdown further increased TF. Hypoxia resulted in decreased CD2AP and actin reorganization in podocytes, and these changes were attenuated by TF siRNA.

Conclusion

Hypoxia increased the expression of TF in human podocytes NF-κB dependently. TF may have a critical role in the hypoxic podocyte injury.

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Authors and Affiliations

  1. Department of Cardiology and Nephrology, Hirosaki University Graduate School of Medicine, 5 Zaifu-cho, Hirosaki, 036-8562, Japan

    Ikuyo Narita, Michiko Shimada, Hideaki Yamabe, Takahiko Kinjo, Tomohiro Tanno, Kimitaka Nishizaki, Misato Kawai, Masayuki Nakamura, Reiichi Murakami, Norio Nakamura, Hirofumi Tomita & Ken Okumura

  2. Bristol Renal, University of Bristol, Dorothy Hodgkin Building, Bristol, UK

    Moin A. Saleem

  3. University of Hong Kong, Hong Kong, China

    Peter W. Mathieson

Authors
  1. Ikuyo Narita
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  2. Michiko Shimada
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  3. Hideaki Yamabe
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  4. Takahiko Kinjo
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  11. Hirofumi Tomita
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  13. Peter W. Mathieson
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  14. Ken Okumura
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Corresponding author

Correspondence to Michiko Shimada.

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Narita, I., Shimada, M., Yamabe, H. et al. NF-κB-dependent increase in tissue factor expression is responsible for hypoxic podocyte injury. Clin Exp Nephrol 20, 679–688 (2016). https://doi.org/10.1007/s10157-015-1214-z

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  • DOI: https://doi.org/10.1007/s10157-015-1214-z

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