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EGCG decreases binding of calcium oxalate monohydrate crystals onto renal tubular cells via decreased surface expression of alpha-enolase

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Abstract

Crystal retention on tubular cell surface inside renal tubules is considered as the earliest and crucial step for kidney stone formation. Therapeutics targeting this step would cease the development of kidney stone. This study thus aimed to investigate the potential role of epigallocatechin-3-gallate (EGCG), a major antioxidant found in green tea leaves, in the reduction of calcium oxalate monohydrate (COM) crystal binding onto renal tubular cells. Pretreatment of the cells with EGCG for up to 6 h significantly diminished crystal-binding capability in a dose-dependent manner. Indirect immunofluorescence assay without and with cell permeabilization followed by laser-scanning confocal microscopy revealed that EGCG significantly reduced surface expression of alpha-enolase, whereas its intracellular level was increased. Western blot analysis confirmed such contradictory changes in membrane and cytosolic fractions of EGCG-treated cells, whereas the total level in whole cell lysate remained unchanged. Moreover, overexpression of surface alpha-enolase and enhancement of cell–crystal adhesion induced by 10 mM sodium oxalate were completely abolished by EGCG. Taken together, these data indicate that EGCG decreases binding of COM crystals onto renal tubular cells by decreasing the surface expression of alpha-enolase via re-localization or inhibition of alpha-enolase shuttling from the cytoplasm to the plasma membrane. These findings may also explain the effects of EGCG in reducing COM crystal deposition in previous animal models of kidney stone disease. Thus, EGCG may be useful for the prevention of new or recurrent stone formation.

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Acknowledgments

We thank Ms. Patcharin Keawboonraung for her technical assistance. This study was supported by the Mahidol University Research Grant (to RK), the Thailand Research Fund (RTA5680004), and the Office of the Higher Education Commission and Mahidol University under the National Research Universities Initiative. NS is supported by the Royal Golden Jubilee Ph.D. Program scholarship, whereas VT and RK are also supported by the “Chalermphrakiat” Grant, Faculty of Medicine Siriraj Hospital.

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Correspondence to Visith Thongboonkerd.

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Kanlaya, R., Singhto, N. & Thongboonkerd, V. EGCG decreases binding of calcium oxalate monohydrate crystals onto renal tubular cells via decreased surface expression of alpha-enolase. J Biol Inorg Chem 21, 339–346 (2016). https://doi.org/10.1007/s00775-016-1344-0

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  • DOI: https://doi.org/10.1007/s00775-016-1344-0

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