银杏叶提取物对高糖环境下人视网膜血管内皮细胞增殖、凋亡的影响

Effects of ginkgo biloba extract on proliferation and apoptosis of human retinal capillary endothelial cells in high glucose environment

目的:研究不同浓度的银杏叶提取物(ginkgo biloba ex-tract,GBE)对体外培养的高糖环境下人视网膜微血管内皮细胞(human retinal capillary endothelial cells,HRCECs)增殖和凋亡的影响。方法:体外培养从角膜移植术后新鲜人眼球提取的HRCECs。取生长良好的第3～4代细胞用于实验,实验分为低糖对照组、高糖对照组、高糖+不同浓度GBE组,用噻唑蓝(MTT)比色法检测HRCECs的增殖。吖啶橙(AO)/溴乙锭(EB)荧光双染色、流式细胞术检测HRCECs的凋亡率,并在荧光显微镜下观察荧光染色后HRCECs凋亡形态学改变。结果:MTT比色法结果显示:用12.5,25.0,50.0,100.0mg/L的GBE处理高糖环境下HRCECs,作用24,48,72h,高糖+不同浓度GBE组与高糖对照组相比具有显著性差异(P<0.05),高糖+不同浓度GBE组之间两两比较均具有显著性差异(P<0.05)。AO/EB荧光染色法、流式细胞术检测细胞凋亡率显示:高糖+不同浓度GBE组与高糖对照组比较均有显著性差异(P<0.05),高糖对照组显著高于低糖对照组(P<0.05)。荧光显微镜下观察到活细胞,早期凋亡细胞,晚期凋亡细胞,非凋亡的死亡细胞生物学形态改变的特征。结论:GBE促进高糖环境下HRCECs的增殖,抑制其凋亡。

AIM： To investigate the effects of ginkgo biloba extract with different concentration on the proliferation and apoptosis of human retinal capillary endothelial cells （ HRCECs）. METHODS： HRCECs from fresher eyeball after keratoplasty were extracted and cultured in vitro, which being at the 3-4rd generation and good growth were taken in experiment. The experimental subjects were divided into low glucose control group, high glucose control group, and high glucose ＋ different concentration GBE groups. The effect of GBE on the proliferation of HRCECs was measured by MTT assay. AO/EB double fluorescence staining method and flow cytometry was used to examine the influence of GBE on the apoptosis of HRCECs. Morphology change on the apoptosis of HRCECs was observed by fluorescence microscope. RESULTS： We observed the effects of GBE on proliferation of HRCECs through MTT assay, which were exposed to 12.5, 25.0, 50.0, 100.0mg/L for 24, 48, 72 hours, respectively. There were significant differences between high glucose control group and high glucose ＋ different concentration GBE groups （ P〈 0.05）, as well as any two groups among high glucose ＋ diffe-rent concentration GBE groups. Apoptosis rate of HRCECs by AO/EB double fluorescence staining method and flow cytometry examination suggested that there were significant differences between high glucose control group and high glucose ＋ different concentration GBE groups were （P〈0.05）, and between high glucose control group and low glucose group （ P 〈 0.05）. It was observed that morphological change on the apoptosis of HRCECs at the different stages was not same. CONCLUSION. GBE promotes proliferation and inhibits apoptosis of HRCECs in the circumstance of high glucose in vitro.