PMID- 15755756
OWN - NLM
STAT- MEDLINE
DCOM- 20050728
LR  - 20220310
IS  - 0931-0509 (Print)
IS  - 0931-0509 (Linking)
VI  - 20
IP  - 5
DP  - 2005 May
TI  - Diphenyleneiodium (DPI) reduces oxalate ion- and calcium oxalate monohydrate and 
      brushite crystal-induced upregulation of MCP-1 in NRK 52E cells.
PG  - 870-8
AB  - BACKGROUND: Our earlier studies have demonstrated upregulation of monocyte 
      chemoattractant protein-1 (MCP-1) in NRK52E rat renal epithelial cells by 
      exposure to oxalate (Ox) ions and crystals of calcium oxalate monohydrate (COM) 
      or the brushite (Br) form of calcium phosphate. The upregulation was mediated by 
      reactive oxygen species (ROS). This study was performed to investigate whether 
      NADPH oxidase is involved in ROS production. METHODS: Confluent cultures of 
      NRK52E cells were exposed to Ox ions or COM and Br crystals. They were exposed 
      for 1, 3, 6, 12, 24 and 48 h for isolation of MCP-1 mRNA and 24 h for 
      enzyme-linked immunosorbent assay (ELISA) to determine the secretion of protein 
      into the culture medium. We also investigated the effect of free radical 
      scavenger, catalase, and the NADPH oxidase inhibitor diphenyleneiodium (DPI) 
      chloride, on the Ox- and crystal-induced expression of MCP-1 mRNA and protein. 
      The transcription of MCP-1 mRNA in the cells was determined using real-time 
      polymerase chain reaction. Hydrogen peroxide and 8-isoprostane were measured to 
      investigate the involvement of ROS. RESULTS: Exposure of NRK52E cells to Ox ions 
      as well as the crystals resulted in increased expression of MCP-1 mRNA and 
      production of the chemoattractant. Treatment with catalase reduced the Ox- and 
      crystal-induced expression of both MCP-1 mRNA and protein. DPI reduced the 
      crystal-induced gene expression and protein production but not Ox-induced gene 
      expression and protein production. CONCLUSIONS: Exposure to Ox ions, and COM and 
      Br crystals stimulates a ROS-mediated increase in MCP-1 mRNA expression and 
      protein production. Reduction in ROS production, lipid peroxidation, low-density 
      lipoprotein release, and inducible MCP-1 gene and protein in the presence of DPI 
      indicates an involvement of NADPH oxidase in the production of ROS.
FAU - Umekawa, Tohru
AU  - Umekawa T
AD  - Department of Pathology and Laboratory Medicine, University of Florida College of 
      Medicine, Box 100275, Gainesville, FL 32610-0275, USA.
FAU - Byer, Karen
AU  - Byer K
FAU - Uemura, Hirotsugu
AU  - Uemura H
FAU - Khan, Saeed R
AU  - Khan SR
LA  - eng
GR  - 059765/PHS HHS/United States
GR  - R01 DK 053962/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, U.S. Gov't, P.H.S.
DEP - 20050308
PL  - England
TA  - Nephrol Dial Transplant
JT  - Nephrology, dialysis, transplantation : official publication of the European 
      Dialysis and Transplant Association - European Renal Association
JID - 8706402
RN  - 0 (Calcium Phosphates)
RN  - 0 (Ccl2 protein, rat)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Onium Compounds)
RN  - 0 (Oxalates)
RN  - 0 (RNA, Messenger)
RN  - 0 (Reactive Oxygen Species)
RN  - 2612HC57YE (Calcium Oxalate)
RN  - 6HJ411TU98 (diphenyleneiodonium)
RN  - EC 1.1.1.27 (L-Lactate Dehydrogenase)
RN  - EC 1.6.3.- (NADPH Oxidases)
RN  - O7TSZ97GEP (calcium phosphate, dibasic, dihydrate)
SB  - IM
MH  - Animals
MH  - Calcium Oxalate/*pharmacology
MH  - Calcium Phosphates/*pharmacology
MH  - Cells, Cultured
MH  - Chemokine CCL2/biosynthesis/*genetics
MH  - Gene Expression Regulation/*drug effects
MH  - L-Lactate Dehydrogenase/metabolism
MH  - Lipid Peroxidation
MH  - NADPH Oxidases/metabolism
MH  - Onium Compounds/*pharmacology
MH  - Oxalates/*pharmacology
MH  - RNA, Messenger/analysis
MH  - Rats
MH  - Reactive Oxygen Species
MH  - Up-Regulation
EDAT- 2005/03/10 09:00
MHDA- 2005/07/29 09:00
CRDT- 2005/03/10 09:00
PHST- 2005/03/10 09:00 [pubmed]
PHST- 2005/07/29 09:00 [medline]
PHST- 2005/03/10 09:00 [entrez]
AID - gfh750 [pii]
AID - 10.1093/ndt/gfh750 [doi]
PST - ppublish
SO  - Nephrol Dial Transplant. 2005 May;20(5):870-8. doi: 10.1093/ndt/gfh750. Epub 2005 
      Mar 8.