The beneficial effect of ginsenosides extracted by pulsed electric field against hydrogen peroxide-induced oxidative stress in HEK-293 cells.
The beneficial effect of ginsenosides extracted by pulsed electric field against hydrogen peroxide-induced oxidative stress in HEK-293 cells.
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脉冲电场提取的人参皂苷对过氧化氢诱导的 HEK-293 细胞氧化应激的有益作用
DOI：
10.1016/j.jgr.2016.02.007
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发表时间：
2017-04
影响因子：
6.3
通讯作者：
Liu J
中科院分区：
文献类型：
--
作者：
Liu D;Zhang T;Chen Z;Wang Y;Ma S;Liu J;Liu J
人参皂苷是人参根的主要药理成分，被认为是抑制氧化应激的主要原因。以2,2-二苯基-1-苦味酰肼基自由基清除能力和氧自由基吸收能力为指标评价人参皂苷的抗氧化活性。将人胚胎肾293 （HEK-293）细胞与脉冲电场（PEF）和溶剂冷浸萃取（SCSE）提取的人参皂苷孵育24 h后，用40μM H2O2诱导细胞损伤。采用MTS法和扫描电镜观察HEK-293细胞的细胞活力和表面形态。采用双乙酸二氯-二氢荧光素荧光探针法测定细胞内活性氧水平。采用HepG2细胞抗氧化活性测定法评价人参皂苷的细胞内抗氧化活性。PEF提取物具有较高的2,2-二苯基-1-苦味酰肼自由基清除能力和较强的氧自由基吸收能力（氧自由基吸收能力值为14.48±4.04μM TE / μg/mL）。HEK-293细胞模型也显示PEF提取物的保护作用呈剂量依赖性。二氯-二氢-双乙酸荧光素实验进一步证明PEF提取物在减少细胞内活性氧积累方面更有活性（比SCSE提取物高8%）。此外，扫描电镜图像显示，PEF提取物处理的HEK-293细胞保持了更完整的表面形态。细胞抗氧化活性值表明，PEF提取的人参皂苷比SCSE提取的人参皂苷具有更强的细胞抗氧化活性。本研究证实了PEF法提取人参皂苷的体外抗氧化作用。此外，PEF可能比SCSE更适用于提取具有增强抗氧化活性的人参皂苷。
Ginsenosides are the main pharmacological components of Panax ginseng root, which are thought to be primarily responsible for the suppressing effect on oxidative stress. 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and oxygen radical absorption capacity were applied to evaluate the antioxidant activities of the ginsenosides. Human embryonic kidney 293 (HEK-293) cells were incubated with ginsenosides extracted by pulsed electric field (PEF) and solvent cold soak extraction (SCSE) for 24 h and then the injury was induced by 40μM H2O2. The cell viability and surface morphology of HEK-293 cells were studied using MTS assay and scanning electron microscopy, respectively. Dichloro-dihydro-fluorescein diacetate fluorescent probe assay was used to measure the level of intracellular reactive oxygen species. The intracellular antioxidant activities of ginsenosides were evaluated by cellular antioxidant activity assay in HepG2 cells. The PEF extracts displayed the higher 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and stronger oxygen radical absorption capacity (with an oxygen radical absorption capacity value of 14.48 ± 4.04μM TE per μg/mL). The HEK-293 cell model also suggested that the protective effect of PEF extracts was dose-dependently greater than SCSE extracts. Dichloro-dihydro-fluorescein diacetate assay further proved that PEF extracts are more active (8% higher than SCSE extracts) in reducing intracellular reactive oxygen species accumulation. In addition, scanning electron microscopy images showed that the HEK-293 cells, which were treated with PEF extracts, maintained more intact surface morphology. Cellular antioxidant activity values indicated that ginsenosides extracted by PEF had stronger cellular antioxidant activity than SCSE ginsenosides extracts. The present study demonstrated the antioxidative effect of ginsenosides extracted by PEF in vitro. Furthermore, rather than SCSE, PEF may be more useful as an alternative extraction technique for the extraction of ginsenosides with enhanced antioxidant activity.