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Hypoxia/re-oxygenation injury induces apoptosis of LLC-PK1 cells by activation of caspase-2

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Abstract

Hypoxia/re-oxygenation injury induces apoptosis in renal tubule cells, but its underlying molecular pathways are not fully elucidated. Activation of caspase-2 has recently been proposed as a novel mechanism of apoptosis in fibroblasts. In this study we examined whether hypoxia/re-oxygenation injury induces apoptosis in proximal tubule cells by activation of caspase-2. Porcine proximal tubule (LLC-PK1) cells were subjected to hypoxia/re-oxygenation injury in the presence or absence of caspase inhibitors. Apoptosis was detected by DNA laddering, flow cytometry, and immunocytochemistry for Bax and cytochrome c. The activity of caspases-2, 8 and 9 was measured. Apoptosis was evident after hypoxia/re-oxygenation and was best prevented by pretreatment with caspase-2 inhibitor. Hypoxia/re-oxygenation resulted in a dramatic increase in caspase-2 activity (32-fold, in comparison with a 16-fold increase in caspase-8 activity and a tenfold increase in caspase-9 activity). Immunocytochemistry revealed Bax activation and translocation to mitochondria and cytochrome c release into the cytosol following hypoxia/re-oxygenation, both of which were significantly suppressed by pretreatment with caspase-2 inhibitor. These results indicate that hypoxia/re-oxygenation injury in cultured proximal tubule cells induced apoptosis by activation of caspase-2, which is required for the mitochondrial translocation of Bax.

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Acknowledgment

Dr. Park is supported by grants from the Samsung Biomedical Research Institute (#SBRI C-A3-211-1) and from the Hyo Seok Research Fund in Kangbuk Samsung Hospital. Dr. Devarajan is supported by grants from the NIH/NIDDK (RO1-DK53289, P50-DK52612, R21-DK070163).

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Authors and Affiliations

  1. Pediatrics, Kangbuk Samsung Hospital, Sungkyunkwan University School of Medicine, Seoul, South Korea

    Moon Soo Park & Beom-Su Kim

  2. Department of Molecular Biology, Chonbuk National University, Chonju, Chonbuk, South Korea

    Beom-Su Kim

  3. Nephrology and Hypertension, Cincinnati Children’s Hospital Medical Center, MLC 7022, 3333 Burnet Avenue, Cincinnati, Ohio, 45229-3039, USA

    Prasad Devarajan

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  1. Moon Soo Park
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  2. Beom-Su Kim
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Correspondence to Moon Soo Park.

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Park, M.S., Kim, BS. & Devarajan, P. Hypoxia/re-oxygenation injury induces apoptosis of LLC-PK1 cells by activation of caspase-2. Pediatr Nephrol 22, 202–208 (2007). https://doi.org/10.1007/s00467-006-0256-6

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  • DOI: https://doi.org/10.1007/s00467-006-0256-6

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