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目的探讨不同浓度的葡萄糖对体外培养的ECV-304细胞增殖和氧化应激的影响。方法体外培养人脐静脉内皮细胞株ECV-304细胞,用浓度为5.5mmol/L(正常对照组)、15mmol/L(HG15组)、25mmol/L(HG25组)、35mmol/L(HG35组)、45mmol/L(HG45组)的葡萄糖培养液分别干预培养48、72和96h。采用MTT法检测不同干预组的细胞增殖能力;硫代巴比妥酸盐法测定细胞中丙二醛(Malondialdehyde,MDA)含量;采用二硫代双硝基苯甲酸比色法测定细胞中还原型谷胱甘肽(glutathione,GSH)含量。结果随着葡萄糖浓度和暴露时间的增加,细胞增殖明显被抑制,细胞内GSH含量显著降低,与正常对照组比较各组细胞间差异有统计学意义(P<0.01)。同时,高浓度葡萄糖组MDA含量显著增加,与正常对照组比较各组细胞间差异有统计学意义(P<0.01)。结论葡萄糖可以明显抑制体外培养的人脐静脉内皮细胞株ECV-304细胞增殖,并造成显著的细胞内氧化应激,并且随着葡萄糖浓度的升高和暴露时间的延长,上述效应呈现出向严重发展的趋势。
Objective To investigate the effects of different concentrations of glucose on the proliferation and oxidative stress of cultured ECV-304 cells in vitro. Methods Human umbilical vein endothelial cell line ECV-304 cells were cultured in vitro. The cells were cultured in the medium containing 5.5 mmol / L of normal saline, 15 mmol / L of HG15, 25 mmol / L of HG25, 35 mmol / L of HG35, , 45mmol / L (HG45 group) glucose culture medium were cultured for 48, 72 and 96h. The proliferation of cells in different intervention groups was detected by MTT assay. Malondialdehyde (MDA) content in cells was detected by thiobarbituronate method. The reduction of cells in the cells was detected by dithiobisnitrobenzoate colorimetry Glutathione (GSH) content. Results With the increase of glucose concentration and exposure time, the cell proliferation was significantly inhibited and the intracellular GSH content was significantly decreased. Compared with the normal control group, there was a significant difference (P <0.01). At the same time, the content of MDA in high concentration glucose group increased significantly compared with that in normal control group (P <0.01). Conclusion Glucose can significantly inhibit the proliferation of human umbilical vein endothelial cell line ECV-304 cultured in vitro and cause significant intracellular oxidative stress. With the increase of glucose concentration and the prolongation of exposure time, the above effects show a serious development the trend of.