银杏叶提取物对糖尿病患者外周血内皮祖细胞GPX及Caspase-3活性的影响

Effects of Ginkgo Biloba Extract on Glutathione Peroxidase and Caspase-3 of Endothelial Progenitor Cells in the Peripheral of Diabetes

目的:观察银杏叶提取物(GBE)对2型糖尿病(T2DM)患者外周血内皮祖细胞(EPCs)中谷胱甘肽过氧化物酶(GPX)及特异性半胱氨酸蛋白酶-3(Caspase-3)表达的影响。方法:取T2DM患者(实验组)和正常人(对照组)外周血分离单个核细胞,在包被有纤维连接蛋白的培养板中加入含血管内皮生长因子(VEGF)、表皮生长因子(EGF)等的M199培养基,诱导EPCs分化,48h后去除悬浮细胞,继续培养,于培养第6天加入不同浓度GBE(0mg/L、6.75mg/L、12.50mg/L、25.00mg/L、50.00mg/L)干预24h后,用紫外分光光度计检测培养上清液中GPX活性,用酶标仪检测培养细胞Caspase-3活性。结果:相同GBE浓度时,实验组GPX活性较对照组明显降低(P<0.05),Caspase-3活性较对照组明显升高(P<0.05或P<0.01)。结论:GBE对T2DM患者EPCs有保护作用,其机制可能与抗氧化和减少自由基生成有关。

Objective: To observe the effects of Ginkgo biloba extract (GBE) on glutathione peroxidase(GPX) and Caspase-3 in endothelial progenitor cells derived from peripheral blood of diabetic patients. Method: The mononuclear cells, isolated from peripheral blood of experimental group and control group, were cultured in Medium 199 with human vascular endothelial growth factor(VEGF) and epidermal growth factor(EGF). After 2 days, non-adherent cells were removed, the culture was maintained. When the 6th days, attached cells were stimulated with different concentrations (0mg/L,6.75mg/L,12.50mg/L,25.00mg/L,50.00mg/L) of GBE for 24 hours, then GPX activity was detected by UV spectrophotometer, Caspase-3 activity was measured by microplate reader. Results: At the same concentration of GBE, compared with the control group, the GPX activity decreased(P<0.05)and the Caspase-3 activity increased(P<0.05 or P<0.01)in the experimental group. Conclusion: GBE has a protective effect on endothelial progenitor cells in diabetes, and its protective mechanism may related to against oxidation and reduction of free radicals.