The Study In Vitro On The Interaction Of Calcium Oxalate Crystals With Ana-1 Macrophage Cell Line In Mouse

Objective To observe the interaction of the calcium oxalate crystals (COM) with macrophages in vitro. To observe and investigate the mechanisms of macrophage cell injury induced by COM, and to explore the potential role of macrophages in the formation of Calcium oxalate kidney stones.Methods Mouse macrophage Ana-1 cells were cultured in vitro and the culture medium was added with COM crystals, the cell-crystal reaction was detected by light microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). To investigate the oxidative stress, NADPH oxidase inhibitor apocynin was chosed as the intervener. We assessed the levels of H2O2, LDH, MDA in the mediums, as well as the cellular ROS and the activity of NADPH oxidase in macrophage cells. The expression of NADPH oxidase subunit p22phox and p47phox mRNA were detected by real-time quantitative PCR(RT-PCR).Results COM crystals can induce changes of the macrophage morphology, we have observed the adhesion and phagocytosis of COM crystals by the macrophages under SEM and TEM; Treatment with apocynin reduced the oxidative stress of macrophages.Conclusion We detected the adhesion and phagocytosis of COM crystals by the macrophages; COM crystals can cause macrophage oxidative stress, Administration of Apocynin can reduce macrophage oxidative stress injury. The results support that the involvement of macrophages in the formation of calcium oxalate kidney stones.