【摘要】 目的:探讨蛹虫草乙酸乙酯提取物(CME)对高糖诱导人脐静脉内皮细胞(HUVECs)衰老的影响。方法:用高糖(40mmol/L)诱导建立HUVECs衰老体外模型,以冬虫夏草提取物(CSE)(50μg/mL)为阳性对照,观察CME(12.5～100μg/mL)对内皮细胞衰老的干预作用。四甲基偶氮唑蓝(MTT)比色法检测细胞增殖;SA-β-半乳糖酐酶(SA-β-gal)染色法检测细胞衰老特异性的SA-β-gal活性;流式细胞术检测细胞周期分布和细胞内活性氧(ROS)含量。结果:经高糖干预24、48、72h后,HUVECs增殖活性OD值明显低于对照组;而且在高糖孵育96h后,SA-β-gal染色阳性细胞率明显增加;孵育48h后,处于G0/G1期细胞的百分率和ROS相对水平明显增加(P<0.01)。与高糖组相比,CME(25～50μg/mL)孵育48和72h后细胞OD值显著提高(P<0.05);CME(12.5～100μg/mL)孵育96h后SA-β-gal染色阳性细胞率显著降低(P<0.01);孵育48h后G0/G1期的细胞百分率(P<0.01)、细胞内ROS水平显著降低(P<0.05);CME 25、50μg/mL效果优于CME 12.5、100μg/mL(P<0.05),且与CSE 50μg/mL作用相近(P>0.05)。结论:蛹虫草乙酸乙酯提取物可促进内皮细胞生长增殖,可能与其降低细胞内ROS水平,减轻细胞氧化应激损伤有关,从而减轻内皮细胞衰老。更多还原

【Abstract】 AIM: To determine the effect of the ethyl acetate extract of cordyceps militaris(CME) on senescence in cultured human umbilical vein endothelial cells(HUVECs) treated with high glucose.METHODS: The HUVECs senescence model was established by incubation with high glucose(40 mmol/L),the extract of Cordyceps Sinensis(CSE)(50 μg/mL) was treated as a positive control,and then CME(12.5-100 μg/mL) was treated to observe its effect on cell senescence.Cell proliferation was determined by the methylthiazolyl tetrazolium(MTT) reduction assay,SA-β-galactosidase(SA-β-gal) positive cells were detected by X-gal,and phase distribution of cell cycles and cellular reactive oxygen species(ROS) were detected by flow cytometry.RESULTS: Compared with unintervented cells,cell OD value significantly decreased in cells treated with high glucose for 24,48,and 72 h(P<0.01).The percentage of cells positive for SA-β-gal,the percentage in G0/G1 phase and ROS levels notably increased after cells incubated by high glucose for 96 or 48 h(P<0.01).Compared with high glucose intervention cells,cell OD value significantly increased in cells intervented by CME(25-50 μg/mL) for 48 h and 72 h(P<0.05);CME(12.5-100 μg/mL)stimulating for 96 h significantly lowered the percentage of cells positive for SA-β-gal(P<0.01);CME(12.5-100 μg/mL)incubation for 48 h significantly lowered the percentage of G0/G1 phase of HUVECs(P<0.01) and ROS production(P<0.05);All these effects of CME 25,50 μg/mL groups were better than those12.5,100 μg/mL groups(P<0.05),and were similar to CSE 50 μg/mL group(P>0.05).CONCLUSION: Ethyl acetate extract of cordyceps militaris may promote the HUVECs growth and proliferation,protecting from high glucose-stimulated vascular endothelial senescence.The effects may be associated with ethyl acetate extract of cordyceps militaris inhibiting reactive oxygen species and alleviating oxidative stress injury.更多还原