草酸诱导HK-2细胞核苷酸结合寡聚化结构域样受体3炎症小体的激活在

草酸诱导HK-2细胞核苷酸结合寡聚化结构域样受体3炎

症小体的激活在草酸钙肾结石形成中的作用及机制

齐士勇;何振;徐勇;张志宏

【期刊名称】《中华实验外科杂志》 【年(卷),期】2017(034)010

【摘要】Objective To observe the mechanism of nucleotide binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome to the formation of calcium oxalate stone after exposure to high concentrate of oxalate.Methods The human kidney proximal tubular epithelial HK-2 cells were cultured and stimulated with different concentrations of soluble oxalate cells.Lactate dehydrogenase (LDH) released assays from cell culture medium and 4',6-dianidino-2-phenylindole (DAPI) staining were used to determine the effect of cellular toxicity and injury after exposure to oxalate.All the cells of oxalate group and control group were incubated with calcium oxalate monohydrate (COM) crystals for 24 h,and cell surface adhesion of crystals

were

observed

using

an

inverted

phase-contrast

microscope.Western blotting and real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) were applied to detect protein level and mRNA quantity of NLRP3,cysteinyl aspartate-specific protease-1 (Caspase-1),and interleukin-1β (IL-1 β) from HK-2 cell lysates after exposure to oxalate.FQ-PCR was applied to analyse mRNA quantity of