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Podocyte hypertrophy precedes apoptosis under experimental diabetic conditions

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Abstract

Podocyte hypertrophy and apoptosis are two hallmarks of diabetic glomeruli, but the sequence in which these processes occur remains a matter of debate. Here we investigated the effects of inhibiting hypertrophy on apoptosis, and vice versa, in both podocytes and glomeruli, under diabetic conditions. Hypertrophy and apoptosis were inhibited using an epidermal growth factor receptor inhibitor (PKI 166) and a pan-caspase inhibitor (zAsp-DCB), respectively. We observed significant increases in the protein expression of p27, p21, phospho-eukaryotic elongation factor 4E-binding protein 1, and phospho-p70 S6 ribosomal protein kinase, in both cultured podocytes exposed to high-glucose (HG) medium, and streptozotocin-induced diabetes mellitus (DM) rat glomeruli. These increases were significantly inhibited by PKI 166, but not by zAsp-DCB. In addition, the amount of protein per cell, the relative cell size, and the glomerular volume were all significantly increased under diabetic conditions, and these changes were also blocked by treatment with PKI 166, but not zAsp-DCB. Increased protein expression of cleaved caspase-3 and cleaved poly (ADP-ribose) polymerase, together with increased Bax/Bcl-2 ratios, were also observed in HG-stimulated podocytes and DM glomeruli. Treatment with either zAsp-DCB or PKI 166 resulted in a significant attenuation of these effects. Both PKI 166 and zAsp-DCB also inhibited the increase in number of apoptotic cells, as assessed by Hoechst 33342 staining and TUNEL assay. Under diabetic conditions, inhibition of podocyte hypertrophy results in attenuated apoptosis, whereas blocking apoptosis has no effect on podocyte hypertrophy, suggesting that podocyte hypertrophy precedes apoptosis.

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Acknowledgments

This work was supported by the Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, the National Research Foundation of Korea (NRF) Grant funded by the Korea government (MSIP) (No. NRF-2011-0030086), and a Grant of the Korean Health Technology R&D Project, Ministry of Health and Welfare, Republic of Korea (HI12C0646).

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All the authors declared no competing interests.

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    Authors and Affiliations

    1. Department of Internal Medicine, College of Medicine, Brain Korea 21 for Medical Science, Severance Biomedical Science Institute, Yonsei University, 134 Shinchon-Dong Seodaemoon-Gu, Seoul, 120-752, Korea

      Sun Ha Lee, Jisun Paeng, Hye-Young Kang, Bo Young Nam, Seonghun Kim, Chan Ho Kim, Mi Jung Lee, Hyung Jung Oh, Jung Tak Park, Seung Hyeok Han, Tae-Hyun Yoo & Shin-Wook Kang

    2. Department of Internal Medicine, College of Medicine, International St. Mary’s Hospital, Catholic Kwandong University, Incheon, Korea

      Sung Jin Moon

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    Sun Ha Lee and Sung Jin Moon contributed equally to this work.

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    Lee, S.H., Moon, S.J., Paeng, J. et al. Podocyte hypertrophy precedes apoptosis under experimental diabetic conditions. Apoptosis 20, 1056–1071 (2015). https://doi.org/10.1007/s10495-015-1134-0

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