Increased expression of monocyte chemoattractant protein-1 (MCP-1) by renal epithelial cells in culture on exposure to calcium oxalate, phosphate and uric acid crystals.
BACKGROUND
During the development of non-infectious kidney stones, crystals form and deposit in the kidneys and become surrounded by monocytes/macrophages (M/M). We have proposed that in response to crystal exposure renal epithelial cells produce chemokines, which attract the M/M to the sites of crystal deposition. We investigated the expression of monocyte chemoattractant protein-1 (MCP-1) mRNA and protein by NRK52E rat renal tubular epithelial cells exposed to calcium oxalate (CaOx), brushite (Br, a calcium phosphate) and uric acid (UA) crystals.


METHODS
Confluent cultures of NRK52E cells were exposed to CaOx, Br or UA at a concentration of 250 micro g/ml (66.7 micro g/cm(2)). They were exposed for 1, 3, 6, 12, 24 and 48 h for isolation of mRNA and 24 h for ELISA to determine the secretion of protein into the culture medium. Since cells are known to produce free radicals on exposure to CaOx crystals we also investigated the effect of free radical scavenger catalase on the crystal induced expression of MCP-1 mRNA and protein.


RESULTS
Exposure of NRK52E cells to the crystals resulted in increased expression of MCP-1 mRNA and production of the chemoattractant. CaOx crystals were most provocative while UA the least. Treatment with catalase had a negative effect on the increased expression of both MCP-1 mRNA and protein, which indicates the involvement of free radicals in up-regulation of MCP-1 production.


CONCLUSIONS
Exposure to both CaOx and calcium phosphate crystals stimulates increased production of MCP-1. Free radicals appear to be involved in this up-regulation. Results indicate that MCP-1, which is often associated with localized inflammation, may be one of the chemokine mediators associated with the deposition of various urinary crystals in the kidneys during kidney stone formation. Because of the small number of experiments performed here, results must be confirmed by more extensive studies with larger sample size.
Umekawa
Tohru
T
Department of Pathology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, FL 32610-0275, USA.
Chegini
Nasser
N
Khan
Saeed R
SR
D001483:Base Sequence; D015415:Biomarkers / Q000032:analysis; D002129:Calcium Oxalate / Q000032:analysis / Q000378:metabolism*; D002130:Calcium Phosphates / Q000032:analysis / Q000378:metabolism*; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000032:analysis / Q000378:metabolism*; D003460:Crystallization; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells; D006801:Humans; D007668:Kidney / Q000166:cytology; D007669:Kidney Calculi / Q000175:diagnosis* / Q000209:etiology; D008969:Molecular Sequence Data; D011336:Probability; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D018709:Statistics, Nonparametric; D014527:Uric Acid / Q000032:analysis*
D015415:Biomarkers; D002130:Calcium Phosphates; D018932:Chemokine CCL2; C485828:alpha-tricalcium phosphate; C485830:tetracalcium phosphate; D002129:Calcium Oxalate; D014527:Uric Acid; C485838:calcium phosphate, monobasic, anhydrous; C020243:calcium phosphate; C485829:calcium phosphate, dibasic, anhydrous