金樱子总黄酮体外抗氧化活性的研究  

Study on Antioxidant Activity of Total Flavonoids from Rosa laevigata Michx

作　　者：文红波[1] 曹运长[1] 吴玉兰[1] 张秋菊[1] 冷超群[1] 

机构地区：[1]南华大学药学与生物科学学院,生化与分子生物学教研室,衡阳421001

出　　处：《天然产物研究与开发》2014年第7期1127-1131,共5页Natural Product Research and Development

基　　金：湖南省自然科学基金项目(06JJ30016);湖南省教育厅青年项目(09B090);衡阳市科技局项目(2009KJ09)

摘　　要：本文研究了金樱子总黄酮（RLTF）体外清除自由基的能力，并观察其对过氧化氢诱导损伤的人脐静脉内皮细胞（HUVEC）抗氧化作用的影响。采用邻苯三酚自氧化法、水杨酸法、DPPH法分别测定RLTF对超氧阴离子自由基（O2）、羟自由基（·OH）、二苯代苦味酰基自由基（DPPH·）的清除率；Hoechst染色法分析各组细胞凋亡情况；MTT法测定细胞活力；分光光度法检测各组细胞中SOD、CAT、GSH-Px活性。结果显示，在20—200μg范围内，随着样品加入量增大，RLTF对·OH和DPPH·的清除率逐渐增大，而对O2的清除率则是先增大后减少；RLTF对O2。的清除作用偏低，对·OH的清除率与芦丁相当，对DPPH·的清除能力则强于芦丁而弱于Vc。Hoechst染色观察结果表明RLTF预处理细胞后，能明显提高细胞的抗凋亡作用。与H2O2损伤组比较，不同浓度RLTF预处理组的细胞活力均显著升高（P〈0．01）；不同浓度RLTF预处理组细胞中的SOD、CAT和GSH-Px活性均明显增加（P〈0．01），且呈剂量依赖性。上述结果表明，RLTF具有较强的自由基体外清除能力，能有效提高氧化损伤HUVEC细胞中SOD、CAT、GSH-Px的酶活性，其可能通过清除细胞内自由基和提高细胞抗氧化酶活性来提高细胞的抗氧化活性。This study aimed to evaluate the antioxidant activity of Rosa laevigata Michx total flavonoids （RLTF） and to investigate the antioxidant effects of RLTF on human umbilical vascular endothelial cells （HUVEC） with oxidative damage induced by hydrogen peroxide （ H2O2 ）. The scavenging effects of RLTF on superoxide anion radical （ O2）, hydroxyl radical （ ·OH）, and 1,1-diphenyl-2-picrylhydrazyl radical （ DPPH· ） were assayed by pyrogallol autoxidation, Fenton reaction and DPPH method,respectively. HUVECs were cultured and induced to oxidative injury by H202 ,cell nuclear morphological changes of HUVEC were observed by inverted fluorescent microscope after Hoeehst 33258 staining, the cell vitality was measured by MTr assay. SOD, CAT and GSH-Px activities of HUVEC were assayed using speetro- photometry. The free radical scavenging experiments indicated that with the concentration increasing from 20 to 200 μg, the scavenging capacities of RLTF on ·OH and DPPH· radicals Were increased gradually,while its O2· clearance activity revealed the tendency of increasing firstly and decreasing afterwards. Hoechst 33258 staining observation showed that RLTF can effectively promote the anti-ap0ptosis activity of HUVEC injured by H2O2 following pre-treated with RLTF for 24 h. The cell vitalities in different RLTF concentration groups were obviously raised compared with those of H2O2 groups （P 〈 0.01 ）. The activities of SOD in 0.24 mg/mL and 0.48 mg/mL RLTF groups, and the activities of CAT and GSH-Px in all RLTF groups were significantly higher than those of H202 oxidative injury group （P 〈0.01 ）. The results showed that RLTF can protect HUVEC from oxidative damage by scavenging O2· , . OH and DPPH · , and by enhancing SOD, CAT and GSH-Px activities of HUVEC.

关 键 词：金樱子 总黄酮 自由基 清除作用 抗氧化酶 

分 类 号：R285[医药卫生—中药学]