Phosphate-Induced Apoptosis in Human Peritoneal Mesothelial Cells in vitro

Title
Phosphate-Induced Apoptosis in Human Peritoneal Mesothelial Cells in vitro
Author(s)
도준영박종원육주민[육주민]류혜명[류혜명]최순연[최순연]마사요 모리시타[마사요 모리시타]박선희[박선희]김찬덕[김찬덕]최지영[최지영]정호영[정호영]김용림[김용림]
Keywords
SMOOTH-MUSCLE-CELLS; STAGE RENAL-DISEASE; INORGANIC-PHOSPHATE; VASCULAR CALCIFICATION; EXTRACELLULAR CALCIUM; HEMODIALYSIS-PATIENTS; DIALYSIS PATIENTS; KIDNEY-DISEASE; MORTALITY; HYPERPHOSPHATEMIA
Issue Date
201106
Publisher
KARGER
Citation
AMERICAN JOURNAL OF NEPHROLOGY, v.34, no.1, pp.77 - 86
Abstract
Background: It has been demonstrated that phosphate uptake through the type III sodium-dependent phosphate co-transporter, Pit-1, induced apoptosis of aortic vascular smooth muscle cells and endothelial cells in vitro. However, the apoptotic effects of high phosphate (HP) level in human peritoneal mesothelial cells (HPMCs) are not known. Methods: To examine whether Pit-1 is expressed in HPMCs, we checked the Western blot assay of immunoreactive Pit-1 and the transcription of Pit-1 by reverse transcriptase PCR. We treated several different phosphate concentrations (1-4 m M) and calcium concentrations (1.8 and 2.8 m M) on HPMCs to assess the effects of concentration. MTT, TUNEL assays, and flow cytometry analysis using Annexin V and propidium iodide were performed to identify cell death and apoptosis. Bax and Bcl-2 by Western blot and caspase-3 activity were evaluated by colorimetric assay. In addition, phosphonoformic acid (PFA) and pan-caspase inhibitor, Z-VAD-FMK, were given to prevent phosphate-induced apoptosis. Results: Pit-1 expression on HPMCs was demonstrated. Apoptosis in HPMCs significantly increased with a high concentration of phosphate in a dose- and time-dependent manner, and was enhanced in the presence of 2.8 m M calcium. HP concentrations significantly decreased the anti-apoptotic Bcl-2/Bax ratio and increased caspase-3 activity. The treatment with PFA and Z-VAD-FMK prevented cell death by HP. Conclusion: Phosphate uptake through Pit-1 induces apoptosis in HPMCs by a caspase-related mechanism. Copyright (C) 2011 S. Karger AG, Basel
URI
http://hdl.handle.net/YU.REPOSITORY/25064http://dx.doi.org/10.1159/000329081
ISSN
0250-8095
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의과대학 > 내과학교실 > Articles
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