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目的:研究常春卫矛乙醇、乙酸乙酯和正丁醇提取物分别对血管内皮细胞ECV-304氧化损伤的保护作用。方法:体外培养ECV-304细胞,以H_2O_2诱导ECV-304细胞氧化损伤为模型,分别将常春卫矛乙醇、乙酸乙酯和正丁醇提取物(终浓度为400,200,100,50,25μg·ml~(-1))作用于细胞24 h后,以MTT法检测细胞存活率;Hochest染色观察细胞凋亡情况;以测定超氧化物歧化酶(SOD)、一氧化氮合酶(NOS)和一氧化氮(NO)的含量评价药物对血管内皮细胞氧化应激的影响。结果:常春卫矛乙醇、乙酸乙酯和正丁醇提取物在25~400μg·ml~(-1)浓度范围内,对ECV-304细胞均无细胞毒作用,其中乙酸乙酯提取物在400μg·ml~(-1)浓度下,对细胞有一定增殖作用(P<0.05);常春卫矛乙酸乙酯(浓度50~400μg·ml~(-1))、正丁醇(浓度100~400μg·ml~(-1))提取物对氧化损伤ECV-304细胞存活率有显著提高,OD值与模型组比较,差异有统计学意义(P<0.05或P<0.01)。常春卫矛乙醇、乙酸乙酯和正丁醇提取物能明显提高氧化损伤ECV-304细胞分泌SOD、NO水平(P<0.05或P<0.01),对NOS水平无影响;常春卫矛乙醇、乙酸乙酯、正丁醇提取物均能抑制氧化损伤细胞凋亡,细胞核形态改善,凋亡率与模型组比较差异有统计学意义(P<0.01)。结论:常春卫矛提取物对H_2O_2诱导血管内皮细胞氧化损伤有一定保护作用。
OBJECTIVE: To study the protective effect of ethanol extract, ethyl acetate and n-butanol extract of vancomycin on endothelial cell ECV-304 oxidative damage respectively. Methods: ECV-304 cells were cultured in vitro, and the oxidative injury of ECV-304 cells induced by H 2 O 2 was taken as the model. The extracts of ethanol extract, ethyl acetate and n-butanol were extracted with 400,200,100,50,25μg · ml ~ 1)) for 24 h, the cell viability was detected by MTT assay; the apoptosis was observed by Hochest staining; the levels of superoxide dismutase (SOD), nitric oxide synthase (NOS) and nitric oxide NO) to evaluate the effect of drugs on oxidative stress in vascular endothelial cells. Results: The ethanol extract, ethyl acetate extract and n-butanol extract showed no cytotoxic effect on ECV-304 cells in the concentration range of 25 ~ 400μg · ml ~ (-1), with ethyl acetate extract at 400μg · (P <0.05). The content of ethyl acetate (50 ~ 400μg · ml -1), n-butanol (100 ~ 400μg · ml -1) could significantly increase the survival rate of ECV-304 cells after oxidative injury. The OD value of ECV-304 cells was significantly higher than that of model group (P <0.05 or P <0.01). Ethanol, ethyl acetate and n-butanol extract of ivy significantly increased the levels of SOD and NO secreted by oxidative injury ECV-304 cells (P <0.05 or P <0.01), but had no effect on NOS level. Ester and n-butanol extracts could inhibit the apoptosis of oxidative injury cells, improve the morphology of the nucleus, and the difference between the apoptosis rate and the model group was statistically significant (P <0.01). CONCLUSION: The extract of Chang-hu-wei Spear has a protective effect on H 2 O 2 -induced oxidative damage of vascular endothelial cells.