野阳合花色苷提取物的抗氧化活性

目的：研究野阳合花色苷提取物的体外抗氧化活性，建立过氧化氢（H2O2）诱导的中国仓鼠肺细胞 （V79-4细胞）氧化损伤模型，初步评价其抗氧化应激作用。方法：采用高效液相色谱-质谱联用分析野阳合花色 苷组成；测定花色苷提取物对2’-联氨-双-3-乙基苯并噻唑啉-6-磺酸（2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)，ABTS）自由基和1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl，DPPH）自由基的清除能力；建 立H2O2诱导V79-4细胞氧化损伤模型，分别采用噻唑蓝、硫代巴比妥酸反应产物和荧光探针2’,7’-二氢二氯荧光黄 双乙酸钠测定花色苷提取物对V79-4细胞氧化损伤的保护作用。结果：野阳合提取物含有4 种花色苷，结合保留时 间和质谱数据等信息，推测分别为矢车菊素-3-半乳糖苷、矢车菊素-3-（6”-香豆酰）葡萄糖苷、芍药色素-3-葡萄 糖苷和芍药色素-3-（6”-香豆酰）葡萄糖苷；花色苷提取物对ABTS＋·和DPPH自由基有良好的清除效果，半抑制 浓度分别是（1.72±0.26） μg/mL和（0.74±0.24） μg/mL，清除能力优于阳性对照水溶性VE；细胞氧化损伤模型 中，50～200 μg/mL的花色苷提取物预处理H2O2诱导V79-4细胞氧化损伤，能明显抑制细胞存活率的下降和脂质过 氧化，并降低损伤后胞内活性氧簇水平。结论：野阳合花色苷提取物具有良好的体外自由基清除能力，并对H2O2 诱导的V79-4细胞氧化损伤具有保护作用。

Antioxidant Properties of Anthocyanins Extract from Habenaria ciliolaris Kranzl

Objective: The aim of this study is to investigate the antioxidant activity in vitro of anthocyanins extracted from the fruits of Habenaria ciliolaris Kranzl and the protective effect of the extract on H2O2-induced oxidative stress in V79-4 cells. Methods: The anthocyanin composition was detected by high performance liquid chromatography-mass spectrometry, and the antioxidant activity was evaluated by using 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assays. Moreover, the protective effect of the extract on H2O2-induced oxidative damage in V79-4 cells was measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl- 2-H-tetrazolium bromide, thiobarbituric acid reactive substances and 2’,7’-dichlorofluorescein diacetate assays. Results: The anthocyanins in the extract were identified as cyanidin 3-galactoside, cyanidin 3-(6”-coumaroyl) glucoside, peonidin 3-galactoside and peonidin 3-(6”-coumaroyl) glucoside by retention time and mass spectral data. The anthocyanins extract exhibited strong ABTS and DPPH free radical scavenging capacity with half maximal inhibitory concentrations of (1.72 ± 0.26) and (0.74 ± 0.24) μg/mL, respectively, which was superior to that of soluble VE. The pretreatment of V79-4 cells with the extract in the ranges of 50-200 μg/mL significantly reduced H2O2-induced cytotoxictiy, potentially inhibited lipid peroxidation and reduced the level of intracellular reactive oxygen species. Conclusion: These findings confirm the in vitro free radical scavenging activity of anthocyanins extracted from H. ciliolaris Kranzl and their cytoprotective effect on H2O2-induced oxidative stress in V79-4 cells.