摘要
目的探讨金雀异黄素(genistein,Gen)对高糖诱导的人视网膜色素上皮(retinalpigment epithelium,RPE)细胞的保护作用。方法培养人RPE细胞,分别以Gen0μmol.L-1、50μmol.L-1、100μmol.L-1、200μmol.L-1的药物浓度作用高糖(33mmol.L-1)培养的RPE细胞48h,噻唑蓝比色法检测细胞活性;流式细胞仪检测细胞周期;激光共聚焦显微镜检测细胞内活性氧(reactive oxygen species,ROS)水平;黄嘌呤氧化酶法测定细胞内超氧化物歧化酶(SOD)活性。结果高糖组细胞活力(0.308±0.022)明显低于正常对照组(0.424±0.014)(P<0.01);高糖+Gen组细胞活力较高糖组升高(P<0.05),且随Gen浓度增大,细胞活力增加更明显。与正常对照组比较,高糖组细胞增殖受到抑制,被阻滞于S期和G2期;与高糖组比较,Gen可减轻高糖对细胞增殖的抑制作用。与正常对照组比较,高糖组细胞内ROS的产生明显增多;Gen以浓度依赖方式抑制高糖组RPE细胞内ROS的产生。与正常对照组(31.04±0.02)比较,高糖组SOD活性(12.32±0.02)显著降低(P<0.01);与高糖组比较,高糖+Gen组SOD活性随Gen浓度的增加而升高(均P<0.05)。结论Gen可保护和修复高糖诱导的人RPE细胞的损伤,其作用可能与抗氧化、增强SOD活性有关。
Objective To investigate the protective effects of genistein on human retinal pigment epithelium （RPE） ceils cultured in high glucose. Methods RPE ceils cultured in high glucose （33mmol · L^-1 ） in vitro were treated with genistein at different concentrations of 0 μmol · L^-1,50 μmol · L^-1, 100 μmol · L^-1,200 μmol · L^-1 for 48 hours respectively. The morphologic changes of ceils were observed by light microscopy. Ceils viability was tested by MTF assay. Cell cycle phase distribution of RPE ceils were measuyed by flow cytometry. The intracellular reactive oxygen species （ROS） was detected by confocal microscopy. The validity of SOD was measured with Xanthine Oxidase Enzymic method. Results Cellular viability of high glucose group （ 0.308 ± 0.022 ） was significantly lower than that of normal control group（ 0.424±0.014 ） （ P 〈0.01 ） ; Cellular viability of high glucose with genistein group was higher than that of high glucose group （ P 〈 0.05 ）, and the higher the concentration of genistein was, the more cellular viability was. Compared with normal control group, cell proliferation of high glucose group was inhibited at S phase and G2 phase. Genistein could decrease the inhibiting effect of high glucose on cell proliferation. The level of intracellular ROS of RPE treated by high glucose was markedly increased compared with control group,and it was inhibited by genistein in a concentration-dependent manner. Contrast with control group （ 31.04 ± 0.02 ）, the validity of SOD of high glucose group（ 12.32 ±0.02） decreased markedly. The validity of SOD was increased with the concentration of genistein （ P 〈 0.05 ）. Conclusion Genistein can protect and repair RPE cells damage, which may have relations with anti-oxidation and the activity of antioxidase SOD.
出处
《眼科新进展》
CAS
2007年第12期888-891,共4页
Recent Advances in Ophthalmology
基金
南京医科大学创新基金资助(编号:CX2001003)~~
