山东医药2017，Vol.57Issue(14)：6-9,4.DOI:10.3969/j.issn.1002-266X.2017.14.002
羟基磷灰石对人肾小管上皮细胞骨桥蛋白表达的影响及机制探讨
Effect of hydroxyapatite on osteopontin expression in human renal tubular epithelial cells
摘要
Abstract
Objective To investigate the effect of hydroxyapatite (HAP) on osteopontin (OPN) expression in human renal tubular epithelial cells (HK-2 cells) and to explore the role of HAP in the formation of kidney stones.Methods The cultured HK-2 cells were divided into the control group,HAP1 group,HAP2 group,HAP3 group and HAP4 group.HK-2 cells in the control group were cultured in the normal culture media,from HAP 1 group to HAP 4 group,we respectively added 200,400,600,400 ng/mL HAP suspension to the culture media.After 12,24,48 and 72 h,the cell activity was measured by CCK-8 assay.In addition,when cultured to 24 hours,RT-qPCR and Western blotting were applied to detect the expression levels of OPN mRNA and protein in human renal tubular epithelial cells.Results With the increase of HAP concentration,the cell viability was decreased gradually,and the cell viability was negatively correlated with the concentrations of HAP (r=-0.566 3,P<0.01).With the prolonged time of HAP,the cell viability was decreased and the cell viability was negatively correlated with the time of HAP (r=-0.796 3,P<0.01).The relative expression levels of OPN mRNA in the normal control group,HAP1 group,HAP2 group,HAP3 group and HAP4 group were 1.151±0.029,1.201±0.004,1.269±0.029,1.179±0.022,and 1.406±0.058,respectively.The OPN mRNA expression of the HAP2 group and HAP4 group was higher than that of the normal control group (all P<0.05).The expression levels of OPN protein in the normal control group,HAP1 group,HAP2 group,HAP3 group and HAP4 group were 0.418±0.010,0.436±0.016,0.512±0.021,0.562±0.016 and 0.627±0.023,respectively.The OPN protein expression of the HAP2,HAP3 and HAP4 groups was higher than that of the normal control group (all P<0.05).Conclusion HAP can damage the renal tubular epithelial cells and up-regulate the expression of OPN,which may be one of the mechanisms involved in the formation of kidney stones.关键词
羟基磷灰石/骨桥蛋白/肾结石/肾小管上皮细胞/细胞损伤/肾钙化斑Key words
hydroxyapatite/osteopontin/kidney calculi/renal tubular epithelial cells/cell damage/renal calcification分类
医药卫生引用本文复制引用
余骏川,刘权,邓耀良,黎承杨,陶芝伟,关晓峰,吴基华,宁鑫,刘云龙,何子奇..羟基磷灰石对人肾小管上皮细胞骨桥蛋白表达的影响及机制探讨[J].山东医药,2017,57(14):6-9,4.基金项目
国家自然科学基金资助项目(81360113). （81360113）
