蛹虫草提取物对高糖诱导内皮细胞衰老的影响

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Go／G1期的细胞百分率（P〈0．01）、细胞内ROS水平显著降低（P〈0．05）；CME25、50／μg／mL效果优于CME12．5、100〉g／mL（P〈0．05），且与CSE50ptg／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。

Effect of the extract of cordyceps militaris on senescence in cultured human umbilical vein endothelial cells treated with high glucose

AIM. To determine the effect of the ethyl acetate extract of cordyceps militaris （CME） on senescence in cultured human umbili- cal 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 trea ted as a positive control, and then CME （12.5- 100/μg/mL） was treated to observe its effect on cell senescence. Cell proliferation was deter- mined by the methylthiazolyl tetrazotium （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 un intervented cells, cell OD value significantly de creased in cells treated with high glucose for 24, 48, and 72 h （P%0.01）. The percentage of cells positive for SA-13-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 interven- tion cells, cell OD value significantly increasedin ceils 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 those 12.5, 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 senes cence. The effects may be associated with ethyl acetate extract of cordyceps militaris inhibiting reactive oxygen species and alleviating oxidative stress injury.