紫萍提取物对过氧化氢诱导内皮细胞氧化损伤保护作用探究.docVIP

紫萍提取物对过氧化氢诱导内皮细胞氧化损伤保护作用探究 　【摘要】 目的研究紫萍提取物是否具有保护内皮细胞免受过氧化氢损伤的作用。方法采用MTT比色法筛选紫萍提取物作用于内皮细胞的安全剂量，建立H2O2对内皮细胞的氧化损伤模型，并研究紫萍提取物的保护作用。最后采用检测试剂盒方法分析紫萍提取物作用后内皮细胞中超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和谷胱甘肽过氧化物酶（GSH-Px）的活性变化。结果紫萍提取物的浓度lt;100 μg/ml，作用时间为24 h时，无明显细胞毒性。以1 mmol/L的H2O2作用于ECV-304细胞1 h造氧化损伤模型，10，25，50 μg/ml的紫萍提取物具有预防氧化损伤作用，可使细胞的存活率由21.98%分别上升至37.49%，51.71%和64.74%，其机理可能是紫萍提取物能够剂量依赖性的增强SOD，CAT和GSH-Px等酶的活性。但是紫萍提取物无治疗氧化损伤的作用。结论紫萍提取物可以有效保护内皮细胞免受氧化损伤。 【关键词】 紫萍 氧化损伤 过氧化氢 超氧化物歧化酶 过氧化氢酶 谷胱甘肽过氧化物酶 Abstract：ObjectiveTo study the protective effect of Spirodela polyrrhiza(L.) Schleid extract (SE) against the damage caused by hydrogen peroxide (H2O2) in human vascular endothelial cells (ECV-304). MethodsNormal ECV-304 cells were respectively incubated with SE 5～400 μg/m for 24h-96h, or H2O2 0.125～10 mmol/L for 1～24 h, the survival rate of cells was determined by tetrazolium assay (MTT). To evaluate the protective effect of SE on ECV-304 injury induced by H2O2, ECV-304 was divided into control, model, and SE treatment groups. In SE treatment group, ECV-304 was incubated with SE 5～50 μg/ml for 24 h，before or after H2O2 1mmol/L (final concentration) was added to ECV-304 in the model group and the SE treatment group for 1 h. The survival conditions of ECV-304 were expressed by the OD values assessed by MTT assay. Furthermore, the activities of SOD, CAT and GSH-Px in ECV-304 cells treated with 10～50μg/ml SE were determined by the corresponding assay kits. Results5，10，25，50 μg/ml SE had no significant cytotoxicity on ECV-304 at 24 h，but 100 μg/ml , 200 μg/ml ,and 400μg/ml SE markedly decreased the survival rate at 24 h. The viability of ECV-304 was significantly inhibited by 1 mmol/L H2O2 at 1h. Pretreated with 10，25，50 μg/ml SE, the survival rate and the activities of SOD, CAT and GSH-Px in cells were increased in a concentration-dependent manner. But SE had no protection on ECV-304 injury induced by H2O2. ConclusionSE has dual effects on the survival rate of ECV-304 in vitro, which is related to