Interaction Of Calcifying Nanoparticles And Calcium Oxalate Crystals With Macrophage-derived Cell Line Ana-1in Mouse

Objective To investigate the interaction of CNP and CaOx monohydrate (COM) crystals with mouse macrophage-derived cell line Ana-1in vitro. To observe the mechanisms of macrophage cell injury induced by CNP and/or COM crystals, and to explore the potential role of macrophages and calcifying nanoparticles in the formation of calcium oxalate kidney stones.Methods Ana-1cells were exposed to5Mcfarlands CNP and/or36μg/ml COM crystals for6h.The interaction was observed by scanning electron microscopy (SEM) and transmission electron microscopy(TEM).We determined the release of lactate dehydrogenase (LDH) as marker of injury, and hydrogen peroxide (H2O2) and malondialdehyde(MDA) as sign of Os. Real time-polymerase chain reaction was performed to determine expression of NADPH oxidase subunits p22phox, p47phox mRNAs.Results macrophages phagocyte COM crystals and/or CNP can be obsersed under TEM and SEM. CNP and/or COM crystals treated Ana-1cells there was enhanced the expression of p47phox、p22phox mRNAs and production of H2O2, MDA and LDH. Conclusions macrophages can adhesion and phagocytic both CNP and COM crystals alone or simultaneously. CNP or COM crystals acting alone results in Ana-1cells damage inducted by Os and NADPH oxidase may be involved. And in concert they act synergistically.