Abstract: TH-PO1065

The Plant 3,4,5-Tri-O-Galloyl Quinic Acid Methyl Ester Inhibits Calcium Oxalate Crystals Growth in a Drosophila Model and Decreases Renal Cell Annexin A1 Surface Expression and Crystal Adhesion

Session Information

Category: Mineral Disease

  • 1204 Mineral Disease: Nephrolithiasis

Authors

  • Abd El-Salam, Mohamed A., Mayo Clinic College of Medicine, Rochester, Minnesota, United States
  • Bastos, Jairo, School of Pharmaceutical Sciences of Ribeirao Preto, University of Sao Paulo, Ribeirao Preto, SP, Brazil
  • Han, Jing jing, Mayo Clinic College of Medicince, Rochester, Minnesota, United States
  • Previdi, Daniel, Faculty of Philosophy, Arts and Sciences, University of Sao Paulo, Ribeirão Preto, SP, Brazil
  • Donate, Paulo, Faculty of Philosophy, Arts and Sciences, University of Sao Paulo, Ribeirão Preto, SP, Brazil
  • Romero, Michael F., Mayo Clinic College of Medicine, Rochester, Minnesota, United States
  • Lieske, John C., Mayo Clinic College of Medicine, Rochester, Minnesota, United States
Background

The design of more effective therapies for urinary stone prevention depends on identifying critical pathogenic steps. Adherence and retention of crystals is one potential early event. Many studies have identified crystal-binding proteins (e.g. annexin A1, heat shock protein 90 (HSP90) and α-enolase) on the apical membranes of renal tubular epithelial cells. Agents that affect expression of these molecules could ameliorate crystal retention.

Methods

The plant metabolite 3,4,5-tri-O-galloyl quinic acid methyl ester (QAME) was prepared by total synthesis and its potential effect on calcium oxalate monohydrate (COM) crystal binding to the surface of Madin-Darby Canine Kidney Cells type I (MDCK I) and crystal growth in a Drosophila melanogaster (fruit fly) Malpighian tubule model were studied. Membrane, cytosolic, and total α-enolase, Annexin A1 and HSP90 levels were evaluated by subcellular fractionation followed by Western blot. Immunofluorescence staining and confocal microscopy were also performed on cultured cells.

Results

Pretreatment of MDCK 1 cells with QAME for up to 6 h significantly diminished crystal-binding in a concentration-dependent manner. QAME significantly reduced surface expression of Annexin A1 by immunofluorescence microscopy, whereas the intracellular level increased. Western blot analysis confirmed these changes in membrane and cytosolic fractions of QAME-treated cells, whereas total cell QAME remained unchanged. The compound also significantly decreased the size and growth of COM crystals induced ex vivo in the Drosophila melanogaster Malpighian tubule model.

Conclusion

These data indicate that QAME decreased binding of COM crystals to cells by decreasing the surface expression of Annexin A1 via changing localization of Annexin A1 from the plasma membrane to the cytosol. Thus, QAME may be useful for the prevention and modulation of stone formation. Further pre-clinical and clinical studies should be done for the use of this compound in urolithiasis.