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 mM) and calcium concentrations (1.8 and 2.8 mM) 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 mM 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.
journal_name
Am J Nephroljournal_title
American journal of nephrologyauthors
Park JW,Yook JM,Ryu HM,Choi SY,Morishita M,Do JY,Park SH,Kim CD,Choi JY,Chung HY,Kim YLdoi
10.1159/000329081subject
Has Abstractpub_date
2011-01-01 00:00:00pages
77-86issue
1eissn
0250-8095issn
1421-9670pii
000329081journal_volume
34pub_type
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