生姜提取物抗氧化应激的实验研究

Experimental study on anti-oxidative stress of ginger extract

目的研究生姜提取物对体外培养的人脐静脉内皮细胞(HUVEC)氧化应激损伤的保护作用。方法溶剂提取法提取生姜天然产物。采用H2O2建立体外培养的HUVEC细胞氧化应激损伤模型,以细胞增殖抑制率为50%(IC50)的H2O2浓度作为后续实验的H2O2浓度。将HUVEC随机分为阳性对照组(硫辛酸浓度分别为0.5、1.0、5.0、10.0μg/ml)和生姜总提取物组(生姜总提取物浓度分别为0.5、1.0、5.0、10.0 mmol/L)。均预培养24 h后,加入1 mmol/L H2O2诱导氧化应激损伤,继续孵育4 h后,采用CCK-8还原法检测细胞存活率。根据上述实验结果,以生姜总提取物抗氧化应激作用最强浓度,确定生姜提取物各组分的实验浓度,进行抗氧化应激实验。结果根据生姜提取物溶剂极性大小,得到总提取物和F1、F2、F3、F4、F5、和F6 6个组分。H2O2诱导的氧化应激损伤细胞模型的IC50为1 mmol/L。随着生姜总提取物浓度的增加,其抗氧化应激能力逐渐增强。各组分亚组中,F5组分的抗氧化应激活性最高。结论生姜提取物具有较强的抗氧化应激作用,可减轻H2O2诱导的HUVEC氧化应激损伤;各提取物中,以F5组分的抗氧化应激活性最强。

Objective To study the protective effects of ginger extract on oxidative stress injuries of human umbilical vein endothelial cells(H UVEC) cultured in vitro. Methods Natural ginger extract was extracted by use of solvent extraction. The oxidativestress injury model of HUVEC cultured in vitro was established with H2 O2. The H2 O2 concentration with a cell proliferation inhibition rate of 50%(IC50) was used as the H2 O2 concentration in subsequent experiments. The HUVECs were randomly divided into a positive control group(the concentration of lipoic acid was 0.5, 1.0, 5.0 and 10.0 μg/ml, respectively) and a total ginger extract group(the concentration of the total ginger extract was 0.5, 1.0, 5.0 and 10.0 mmol/L, respectively). After 24 hours of preculture, H2 O2(1 mmol/L) was added to induce oxidative stress injuries. After another four hours of incubation, CCK 8 reduction method was taken to detect the survival rate of cells. The experimental results showed that anti-oxidative stress experiment might be conducted with the experimental concentrations of various components of ginger extract determined on the basis of the concentration of the total ginger extract with the strongest anti-oxidative stress effect. Results The total ginger extract and six components, i.e. F1, F2, F3, F4, F5 and F6, were obtained according to the solvent polarity of ginger extract. The IC50 of the oxidative stress injury cell model induced by H2 O2 was 1 mmol/L. The anti-oxidative stress capacity of the total ginger extract gradually increased with the increase of its concentration.Among the six component sub-groups, the anti-oxidative stress activity in group F5 was the strongest. Conclusion Ginger extract has strong anti-oxidative stress effects and can alleviate the HUVEC oxidative stress injury induced by H2 O2. The anti-oxidative stress activity in group F5 is the strongest among all extract components.