| Superoxide from NADPH oxidase as second messenger for the expression of osteopontin and monocyte chemoattractant protein-1 in renal epithelial cells exposed to calcium oxalate crystals. | |
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MedLine Citation:
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PMID: 19220254 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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OBJECTIVE To test the hypothesis that exposure of a renal epithelial cell line, NRK52E, to calcium oxalate monohydrate crystals (COM) would up-regulate NADPH oxidase subunit p47(phox), enhance superoxide production and increase monocyte chemoattractant protein-1 (MCP-1) and osteopontin mRNA levels. MATERIALS AND METHODS Confluent cultures of NRK52E cells were exposed to COM (66.7 microg/cm(2)) with or with no pretreatment with diphenileneiodium chloride (DPI, 10 x 10(-6)m) an inhibitor for NADPH oxidase, under serum-free conditions. The conditioned medium was collected and total cellular RNA isolated from the cells, and subjected to enzyme-linked immunosorbent assay and real-time polymerase chain reaction (PCR). Production of reactive oxygen species (ROS) was estimated by dihydroethidium (DHE) staining using a fluorescence microscope. Immunohistochemistry and real-time PCR were used to analyse p47(phox) in NRK52E cells. RESULTS In COM treated NRK52E cells there was enhanced expression of p47(phox) and production of superoxide. COM-induced production of MCP-1 and osteopontin was significantly reduced after treatment with DPI. CONCLUSIONS While the generation of a lot of ROS might play a major role in tissue injury or death, the regulated generation of low concentration of ROS, possibly by NADPH oxidase, may represent a second messenger system for generation of COM-induced MCP-1 and osteopontin production in the renal tubules. |
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Authors:
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Tohru Umekawa; Hidenori Tsuji; Hirotsugu Uemura; Saeed R Khan |
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Publication Detail:
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Type: Journal Article Date: 2009-02-10 |
Journal Detail:
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Title: BJU international Volume: 104 ISSN: 1464-410X ISO Abbreviation: BJU Int. Publication Date: 2009 Jul |
Date Detail:
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Created Date: 2009-08-03 Completed Date: 2009-09-02 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 100886721 Medline TA: BJU Int Country: England |
Other Details:
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Languages: eng Pagination: 115-20 Citation Subset: IM |
Affiliation:
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Department of Urology, Kinki University School of Medicine, Osaka, Japan. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Calcium Oxalate / pharmacology Cell Line Chemokine CCL2 / metabolism* Enzyme-Linked Immunosorbent Assay Epithelial Cells Humans Immunohistochemistry Kidney Calculi / etiology, metabolism* Kidney Tubules / metabolism NADPH Oxidase / metabolism* Osteopontin / metabolism* RNA, Messenger / metabolism Rats Reverse Transcriptase Polymerase Chain Reaction Second Messenger Systems / physiology* Superoxides / metabolism* Up-Regulation / physiology |
| Chemical | |
Reg. No./Substance:
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0/Chemokine CCL2; 0/RNA, Messenger; 106441-73-0/Osteopontin; 11062-77-4/Superoxides; 25454-23-3/Calcium Oxalate; EC 1.6.3.1/NADPH Oxidase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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