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Enhanced glycogen synthase kinase-3β activity mediates podocyte apoptosis under diabetic conditions

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Abstract

Glycogen synthase kinase-3β (GSK-3β) is involved in the pathogenesis of various kidney diseases. This study was undertaken to examine the changes in GSK-3β activity in podocytes under diabetic conditions and to elucidate the functional role of GSK-3β in podocyte apoptosis. In vivo, 32 rats were injected with either diluent (n = 16, C) or with streptozotocin intraperitoneally (n = 16, DM), and 8 rats from each group were treated with 6-bromoindirubin-3′-oxime (BIO) for 3 months. In vitro, immortalized mouse podocytes were exposed to 5.6 mM glucose or 30 mM glucose (HG) with or without 10 μM BIO. Western blot analysis and TUNEL or Hoechst 33342 staining were performed to identify apoptosis. Urinary albumin excretion was significantly higher in DM rats, and this increase was significantly abrogated in DM rats by BIO treatment. The protein expression of Tyr216-phospho-GSK-3β was significantly increased in DM glomeruli and in cultured podocytes exposed to HG. Western blot analysis revealed that the protein expression of Bax and active fragments of caspase-3 were significantly increased, whereas phospho-Akt, β-catenin, and Bcl-2 protein expression were significantly decreased in DM glomeruli and HG-stimulated podocytes. Apoptosis, determined by TUNEL assay and Hoechst 33342 staining, was also significantly increased in podocytes under diabetic conditions. The changes in the expression of apoptosis-related molecules and the increase in the number of apoptotic cells in DM glomeruli as well as in HG-stimulated podocytes were significantly ameliorated by BIO. These findings suggest that enhanced GSK-3β activity within podocytes under diabetic conditions is associated with podocyte loss in diabetic nephropathy.

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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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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Internal Medicine, Severance Biomedical Science Institute, BK21, College of Medicine, Yonsei University, 134 Shinchon-Dong, Seodaemoon-Gu, Seoul, 120-752, Korea

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

  2. Department of Internal Medicine, Graduate School of Medicine, Gachon University Gil Medical Center, Gachon University, Incheon, Korea

    Jae Hyun Chang

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  1. Jisun Paeng
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Correspondence to Shin-Wook Kang.

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J. Paeng and J. H. Chang have contributed equally to this work.

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Paeng, J., Chang, J.H., Lee, S.H. et al. Enhanced glycogen synthase kinase-3β activity mediates podocyte apoptosis under diabetic conditions. Apoptosis 19, 1678–1690 (2014). https://doi.org/10.1007/s10495-014-1037-5

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