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Protective role of heparin/heparan sulfate on oxalate-induced changes in cell morphology and intracellular Ca2+

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Abstract

Alterations in intracellular Ca2+ ([Ca2+]i) are generally associated with cellular distress. Oxalate-induced cell injury of the renal epithelium plays an important role in promoting CaOx nephrolithiasis. However, the degree of change in intracellular free calcium ions in renal epithelial cells during oxalate exposure remains unclear. The aim of this study is to determine whether acute short-term exposure to oxalate produces morphological changes in the cells, induces a change in cytosolic Ca2+ levels in renal tubular epithelial cells and whether the application of extracellular glycosaminoglycans (GAGs) prevents these changes. Cultured Mardin-Darby canine kidney cells were exposed to oxalate, and changes in cytosolic Ca2+ were determined under various conditions. The effect of heparin and heparan sulfate (HS) during oxalate exposure was examined. The change in the GAG contents of the culture medium was also determined. Transmission electron microscopy (TEM) was performed for morphological analysis. The degree of change in cytosolic Ca2+ strongly correlated with oxalate concentration. Cytosolic Ca2+ levels decreased in parallel with an increase in the concentration of oxalate. However, this decrease was strongly inhibited by pretreatment with heparin or HS. TEM revealed cytoplasmic vacuolization, the appearance of flocculent material and mitochondrial damage after oxalate exposure. On the other hand, pretreatment with heparin or HS completely blocked these morphological changes. The present data suggest that acute exposure to a high concentration of oxalate challenges the renal cells, diminishes their viability and induces changes in cytosolic Ca2+ levels. Heparin and HS, which are known as potent inhibitors of CaOx crystallization, may also prevent oxalate-induced cell changes by stabilizing the cytosolic Ca2+ level.

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Acknowledgments

This work was supported in part by a Grant-in-Aid for Encouragement of Young Scientists (S.I., M.I.:11770915, 11770916), (S.N.: 12307072) from the Ministry of Education, Science, Sports, and Culture in Japan. We thank Ms. Satoko Yamada for her help in preparing the cells for the transmission electron microscopic study.

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Authors and Affiliations

  1. Department of Urology, Kurume University School of Medicine, 67 Asahi-machi, Kurume 830–0011, Japan

    Shizuka Iida, Shyumei Chikama, Michiro Inoue, Kei Matsuoka & Shinshi Noda

  2. Department of Physiology, Kurume University School of Medicine, Kurume, Japan

    Masaru Ishimatsu & Takashi Akasu

  3. Department of Pathology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, USA

    Saeed R. Khan

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  1. Shizuka Iida
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  2. Masaru Ishimatsu
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  3. Shyumei Chikama
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Correspondence to Shizuka Iida.

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Iida, S., Ishimatsu, M., Chikama, S. et al. Protective role of heparin/heparan sulfate on oxalate-induced changes in cell morphology and intracellular Ca2+ . Urol Res 31, 198–206 (2003). https://doi.org/10.1007/s00240-003-0317-2

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  • DOI: https://doi.org/10.1007/s00240-003-0317-2

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