非洲绿猴肾上皮细胞的损伤模型及其对草酸钙生长的调控作用

Injury Model of African Green Monkey Kidney Epithelial Cell and Its Modulation on Calcium Oxalate Formation

采用H2O2对非洲绿猴肾上皮细胞(Vero)进行了损伤,通过检测细胞存活率、培养基中超氧化物歧化酶(SOD)浓度、丙二醛(MDA)释放量和细胞表面晶体粘附分子骨桥蛋白(OPN)的表达量变化,检测了细胞的损伤程度.H2O2对Vero细胞的损伤作用呈现时间依赖性和浓度依赖性;细胞损伤后,MDA释放量增加,SOD浓度降低,OPN表达量显著增加,导致粘附的晶体量增加.利用扫描电子显微镜(SEM)和X射线衍射(XRD)研究了细胞损伤前后对草酸钙(CaOxa)晶体生长的调控作用.对照组细胞诱导生成的CaOxa晶体棱角圆钝,同时含有一水草酸钙(COM)和二水草酸钙(COD)晶体;而损伤细胞诱导生成的晶体形状不规则,棱角尖锐,主要为COM晶体,因此,细胞损伤后增加了草酸钙结石形成的危险性.所建立的Vero细胞氧化损伤模型有助于从细胞水平上阐明草酸钙结石的形成机制.

H2O2 was used to injure African green monkey renal epithelial cells （Vero）. The injury degree of Vero cells was characterized by detecting the cell survival rate, the concentration change of superoxide dismutase （SOD） in culture medium, the release of malondialdehyde （MDA） and the change of expression quantity of osteopontin （OPN） on cellular surface. The injured degree of Vero cells by H2O2 was time-dependent and concentration-dependent. After injured, the release quantity of MDA increased, the concentration of SOD decreased and the expression quantity of OPN increased significantly, thus resulting in an increase of the adherent quantity of crystals. The modulation of cells before and after injury to the growth of calcium oxalate （CaOxa） crystals was investigated by scanning electron microscopy （SEM） and X-ray diffraction （XRD）. The CaOxa crystals induced by the cells in control group contained simultaneously calcium oxalate monohydrate （COM） and dihydrate （COD） crystals, and these crystals had obtuse edges and comers. However, the crystals induced by the cells in injured group were mainly COM crystals, and they had irregular shape with sharp edges and comers. Consequently, the risk of CaOxa stones formation increased after the cells were injured. The oxidative injury model of Vero cells established in this paper was in favor of understanding the formation mechanism of calcium oxalate stones in cell level.