黄酮对体外培养人角膜内皮细胞的影响及其抗氧化作用

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。

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

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, 600μmol/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 85μmol/L and 100μmol/L flavone could not.CONCLUSION: Flavone has prominent antioxidative acti-vities on HCEC from oxidant induced apoptosis at concen-tration of 55-70 μmol/L, but has obvious apoptosis-inducing effects on HCEC at concentration higher than 85 μmol/L.