黄酮对体外培养人角膜内皮细胞的影响及其抗氧化作用

Effects and antioxidative activities of flavone on human corneal endothelial cells in vitro

目的：确定黄酮对人角膜内皮细胞（human corneal endothelial cells,HCEC）的影响及其对因氧化损伤所致细胞凋亡的抗氧化保护作用。方法：利用体外培养的HCEC,通过添加不同浓度的黄酮继续培养,或经不同浓度黄酮预处理后再添加过氧化氢（H2O2）继续培养,采用光镜形态观察、吖啶橙/溴化乙锭（AO/EB）双染色和DNA琼脂糖凝胶电泳等技术手段,研究黄酮对HCEC的影响及其抗氧化作用。结果：黄酮浓度高于85μmol/L时可诱导HCEC发生细胞凋亡,而低于70μmol/L时对HCEC没有影响;55和70μmol/L黄酮预处理对600μmol/LH2O2所引起的HCEC细胞凋亡具有显著的抑制作用（60%→22%,8%;P〈0.01）,而85和100μmol/L黄酮预处理对H2O2所引起的HCEC细胞凋亡没有显著的抑制作用。结论：55～70μmol/L黄酮对HCEC具有显著的抗氧化保护作用,而浓度高于85μmol/L的黄酮反而能诱发HCEC发生细胞凋亡。

AIM：To evaluate the effect and antioxidative activities of flavone on human corneal endothelial cells（HCEC） in vitro. METHODS： Different concentrations of flavone were added into the culture medium of HCEC, and the morphology of HCEC was observed and their apoptosis was verified by acridine orange/ethidium bromide （AO/EB） double fluorescent staining and DNA agarose gel electrophoresis. After different concentrations of flavone were added into the culture medium of HCEC and cultured for 30 minutes, 600wmol/L hydrogen peroxide （H2O2） was further added into the culture medium of HCEC, and the morphology of HCEC was observed and their apoptosis was verified by AO/EB double fluorescent staining and DNA agarose gel electrophoresis. RESULTS： Flavone had no injury effects on HCEC at low concentrations （lower than 70μmol/L ）, while had obvious apoptosis-inducing effects to HCEC at high concentrations （higher than 85μmol/L）. Preincubation with 55μmol/L and 70μmol/L flavone could inhibit significantly the apoptosis of HCEC induced by 600μmol/L H2O2, while preincubation with 85μ mol/L and 100μmol/L flavone could not. CONCLUSION.. Flavone has prominent antioxidative activities on HCEC from oxidant induced apoptosis at concentration of 55-70μmol/L, but has obvious apoptosis-inducing effects on HCEC at concentration higher than 85 μmol/L.