Exposure of Madin-Darby canine kidney (MDCK) cells to oxalate and calcium oxalate crystals activates nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase

Urology
Aslam KhanSaeed R Khan

Abstract

To investigate nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase activity in Madin-Darby canine kidney (MDCK) cells and the production of reactive oxygen species on exposure to oxalate (Ox) or calcium oxalate (CaOx) crystals. Monolayers of confluent Madin-Darby canine kidney cells were exposed to 100, 300, 500 μmol, 1 mmol Ox or 33, 66, 132 μg/cm(2) CaOx crystals for 15 minutes, 30 minutes, 1 hour, 2 hours, or 3 hours. After specified periods of exposure to Ox and CaOx crystals, lactate dehydrogenase release, trypan blue exclusion, activation of NADPH oxidase, and superoxide production were determined using standard procedures. The production of Nox4, a membrane associated subunit of the NADPH oxidase enzyme, was determined by western blot analysis. Exposure to Ox and CaOx crystals leads to time- and concentration-dependent activation of NADPH oxidase. Western blot analysis showed an increase in the production of Nox4. The production of superoxide also changed in a time- and concentration-dependent manner, with maximum increases after 30-minute exposure to the highest concentrations of Ox and CaOx crystals. Longer exposures did not change the results or resulted in decreased activities. Exposure to higher concentratio...Continue Reading

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Citations

Sep 12, 2015·Materials Science & Engineering. C, Materials for Biological Applications·Xin-Yuan SunQiong-Zhi Gan
Jan 30, 2015·Clinical Science·Chet E HoltermanChris R J Kennedy
Mar 7, 2015·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Xin-Yuan SunQiong-Zhi Gan

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