过氧化氢对人晶状体上皮细胞增生及VEGF表达的影响

目的:研究不同浓度的过氧化氢(H2O2)对体外培养的人晶状体上皮细胞增生及VEGF表达的影响。方法:体外培养的人晶状体上皮细胞系SRA01/04,在培养液中分别加入浓度为1,10,100nmol/L;1,10,100μmol/L和1mmol/L的过氧化氢处理40min,对照组无过氧化氢,随后在正常培养液中继续培养,在0,6,12和24h用MTT方法检测晶状体上皮细胞的增生活力;在24h用ELISA方法检测培养液上清中血管内皮生长因子(VEGF)的含量。结果:在培养液中加入1nmol/L~10μmol/L浓度的H2O2处理组,晶状体上皮细胞增生率(A值)随浓度较对照组明显增加,其中以10nmol/L最为明显;而100μmol/L,1mmol/L处理组,晶状体上皮细胞出现生长抑制。ELISA检测结果显示,1nmol/L~10μmol/L的过氧化氢刺激后,VEGF水平均高于对照组,且呈浓度依赖性,峰值出现在10nmol/L和10μmol/LH2O2处理组。结论:低浓度(1nmol/L~10μmol/L)的H2O2可以促进晶状体上皮细胞增殖,并使VEGF分泌增加;提示VEGF可能作为一种氧化应激产物,对晶状体上皮细胞起到保护、免受损伤的作用。

Effects of hydrogen peroxide on the proliferation and VEGF expression of human lens epithelial cells

AIM:To investigate the effects of hydrogen peroxide(H2O2)at various concentrations on the proliferation and VEGF expression of human lens epithelial cells in vitro.· METHODS:Human epithelial cells SRA01/04 were used.The cells were treated with H2O2 at various concentrations of 1nmol/L,10nmol/L,100nmol/L,1μmol/L,10μmol/L,100μmol/L and 1mmol/L respectively.The controls were cultured in the medium without H2O2.After treated for 40 minutes,the cells were then cultured in normal condition for additional 24 hours.The survival and proliferation of the cells was quantified by MTT assay before treatment and at 6,12,24 hours after the treatment of H2O2.The concentration of VEGF in the supernatants was measured by ELISA at 24 hours after the treatment of H2O2.· RESULTS:The proliferation rates(A values)of the human lens epithelial cells treated with 1nmol/L to 10μmol/L of hydrogen peroxide increased significantly compared to that of the controls,in a dose-dependent manner.Of them,the highest A value was in the cells treated with 10nmol/L H2O2.However,high dose(100μmol/L and 1mmol/L)of hydrogen peroxide suppressed the proliferation of the cells.ELISA test showed that the VEGF levels were elevated also in a dose dependent manner at low dose(1nmol/L-10μmol/L)of hydrogen peroxide.The peak level of VEGF appeared in the cells treated with 10nmol/L and 10μmol/L of H2O2.· CONCLUSION:Hydrogen peroxide at low dose may stimulate the proliferation of lens epithelial cells,and increase production of VEGF,indicating that VEGF can be a product of oxidative stress and serve protection for lens epithelial cells from damage.