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中国精品科技期刊2020
张丽明,马雅鸽,牛若楠,等. 宾川葡萄籽原花青素纯化及对HepG2细胞增殖的影响[J]. 食品工业科技,2022,43(5):230−236. doi: 10.13386/j.issn1002-0306.2021070177.
引用本文: 张丽明,马雅鸽,牛若楠,等. 宾川葡萄籽原花青素纯化及对HepG2细胞增殖的影响[J]. 食品工业科技,2022,43(5):230−236. doi: 10.13386/j.issn1002-0306.2021070177.
ZHANG Liming, MA Yage, NIU Ruonan, et al. Purification of Proanthocyanidins from Binchuan Grape Seed and Its Effects on Proliferation of HepG2 Cells[J]. Science and Technology of Food Industry, 2022, 43(5): 230−236. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021070177.
Citation: ZHANG Liming, MA Yage, NIU Ruonan, et al. Purification of Proanthocyanidins from Binchuan Grape Seed and Its Effects on Proliferation of HepG2 Cells[J]. Science and Technology of Food Industry, 2022, 43(5): 230−236. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021070177.

宾川葡萄籽原花青素纯化及对HepG2细胞增殖的影响

Purification of Proanthocyanidins from Binchuan Grape Seed and Its Effects on Proliferation of HepG2 Cells

  • 摘要: 目的:研究云南大理宾川葡萄籽原花青素纯化及对肝癌HepG2细胞增殖的影响。方法:考察了上样液质量浓度、pH、流速、乙醇体积分数等对葡萄籽原花青素吸附及解析的影响,研究AB-8型大孔吸附树脂纯化云南大理宾川葡萄籽原花青素的条件。将纯化过程中乙醇梯度洗脱得到的不同极性葡萄籽原花青素作用于HepG2细胞,采用CCK8法检测不同浓度各极性葡萄籽原花青素对肝癌HepG2细胞增殖的影响。结果:AB-8树脂纯化葡萄籽原花青素的最佳工艺为上样液pH4.0,质量浓度为6.0 mg/mL,流速为1.0 BV/h,洗脱液的体积分数为30%,洗脱体积4 BV。此纯化使葡萄籽提取物中原花青素的含量从22.77%±0.32%提高至94.73%±0.6429%,回收率为80.55%±1.6499%。以10%、20%、30%、40%乙醇洗脱的葡萄籽原花青素极性部位各浓度对肝癌HepG2细胞的增殖均有抑制作用,其中未纯化部位(即提取物未经过大孔树脂纯化)给药浓度为200 μg/mL时抑制作用最为显著(P<0.001),10%乙醇组IC50为25.09 μg/mL,抑制效果良好。结论:云南大理宾川葡萄籽原花青素用AB-8型大孔树脂进行纯化,方法可行,该葡萄籽原花青素对肝癌HepG2细胞的增殖有较好抑制作用。

     

    Abstract: Objective: To study the purification of grape seed proanthocyanidins from Binchuan, Dali, Yunnan and their effects on the proliferation of HepG2 cells. Method: The effects of sample concentration, pH, flow rate, and ethanol volume fraction on the adsorption and resolution of grape seed proanthocyanidins were investigated, and the conditions for the AB-8 type macroporous adsorption resin to purify grape seed proanthocyanidins from Binchuan, Dali, Yunnan were studied. Grape seed proanthocyanidins of different polarities obtained by the ethanol gradient elution during the purification process were applied to HepG2 cells, and the effects of different concentrations of grape seed proanthocyanidins of different polarities on the proliferation of liver cancer HepG2 cells were detected by the CCK8 method. Results: The best process for AB-8 resin to purify grape seed proanthocyanidins was the pH of the sample solution 4.0, the mass concentration was 6.0 mg/mL, the flow rate was 1.0 BV/h, the volume fraction of the eluent was 30%, and the elution volume was 4 BV. This purification increased the content of proanthocyanidins in the grape seed extract from 22.77%±0.32% to 94.73%±0.6429%, with a recovery rate of 80.55%±1.6499%. The polar parts of grape seed proanthocyanidins eluted with 10%, 20%, 30% and 40% ethanol had inhibitory effects on the proliferation of liver cancer HepG2 cells, and the unpurified part (that is, the extract had not been purified by macroporous resin) that inhibitory effect was most significant when the administration concentration was 200 μg/mL (P<0.001). The IC50 of the 10% ethanol group was 25.09 μg/mL, which showed a better inhibitory effect. Conclusion: The grape seed proanthocyanidins in Binchuan, Dali, Yunnan are purified with AB-8 type macroporous resin, and the method is feasible. The grape seed proanthocyanidins have a good inhibitory effect on the proliferation of liver cancer HepG2 cells.

     

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