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The Study in Vitro on the Interaction of Calcium Oxalate Crystals with Ana-1 Macrophage Cell Line in Mouse

Author: SunBingHua
Tutor: DengYaoLiang
School: Guangxi Medical University
Course: Department of Urology
Keywords: Macrophages Kidney stones NADPH oxidase Reactive oxygen species
CLC: R692.4
Type: Master's thesis
Year: 2009
Downloads: 62
Quote: 0
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Abstract


Objective To observe the macrophages in vitro cell culture system with calcium oxalate monohydrate (calcium oxalate monohydrate, COM) crystal interactions , to observe and to explore the mechanisms of the COM crystal macrophages injury of macrophage formation of calcium oxalate kidney stones the possible role . Methods Cultured mouse macrophage cell line Ana-1, COM co-cultured , and NADPH oxidase inhibitor apocynin intervention treatment . Using light microscopy, scanning and transmission electron microscopy to observe the interaction between macrophages and COM crystal ; cells were divided into four groups according to the purpose of the experiment , each group after 6h colorimetric detection indicators of oxidative stress in the cell culture medium level , and at the same time detection of intracellular reactive oxygen species (reactive oxygen species, ROS) content and NADPH oxidase activity , using real-time quantitative PCR (RT-PCR) method to detect cells in each group NADPH oxidase subunit p22phox and p47phox mRNA expression levels . Results of COM crystals can cause morphological changes of macrophages by scanning transmission electron microscopy to macrophage adhesion , phagocytosis phenomenon of COM crystals ; the COM crystal can stimulate macrophage NADPH oxidase subunit p22phox , p47phox mRNA expression was significantly increased , accompanied by cellular oxidative stress (oxidative stress, OS) injury , apocynin intervention reduced the gene expression , the cells from oxidative stress injury is also reduced . Conclusions with adhesion of macrophages , phagocytic ability of COM crystals ; the COM crystal can stimulate macrophage NADPH oxidase expression and activity increased , causing oxidative stress injury . The results support the participation of macrophages calcium oxalate kidney stone formation process inference .

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CLC: > Medicine, health > Surgery > Urology ( urinary and reproductive system diseases) > Kidney disease > Kidney stones
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