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目的通过灰色关联度分析技术及对Caco-2肠癌细胞增殖情况的影响来评价荆芥总黄酮大孔吸附树脂纯化工艺。方法以总黄酮静态吸附量和解吸率为考察指标筛选大孔吸附树脂型号,采用单因素试验,优化其纯化总黄酮的工艺条件,并采用MTT法测定Caco-2细胞抑制率,同时应用灰色关联度分析技术,计算总黄酮质量分数与对肿瘤细胞增殖抑制率的关联度。结果 HPD-400型大孔吸附树脂对荆芥总黄酮有较好的吸附分离性能,荆芥药材以0.1 g/m L的生药质量浓度、0.8 g/m L(药材/湿树脂)的上柱量上柱,用2倍柱体积(BV)水洗除杂,用8 BV、70%乙醇进行洗脱,体积流量均为4 BV/h,收集洗脱液,挥干,同法进行第2次纯化,总黄酮质量分数为92.15%,对Caco-2细胞增殖抑制率为86.17%,总黄酮质量分数与抑制率灰色关联系数为0.950 2。结论此工艺条件稳定可行,体外抗Caco-2肿瘤细胞药效明确,为开发以荆芥总黄酮纯化物为主成分的抗癌新药奠定基础。
Objective To evaluate the purification technology of macroporous adsorption resin of Nepeta total flavonoids by the gray relational analysis technique and its effect on the proliferation of Caco-2 colon cancer cells. Methods The macroporous resin was screened based on the static adsorption capacity and desorption rate of total flavonoids. The single factor experiment was used to optimize the conditions for the purification of total flavonoids. The inhibition rate of Caco-2 cells was determined by MTT assay and the gray relation Degree of analysis of technology to calculate the total flavonoid content and tumor cell proliferation inhibition rate of relevance. Results HPD-400 macroporous resin has good adsorption and separation performance on the total flavonoids of Schizonepeta tenuifolia. The crude drug of Nepeta was loaded on the column at a crude drug concentration of 0.1 g / m L and 0.8 g / m L (medicinal material / wet resin) The volume of the column, with 2 times the volume of (BV) washed with water, with 8 BV, 70% ethanol elution, the volume flow were 4 BV / h, the eluent was collected, evaporated to dryness, Purified, the total flavonoid mass fraction was 92.15%, the inhibition rate of Caco-2 cell proliferation was 86.17%, and the gray correlation coefficient of total flavonoid mass fraction and inhibition rate was 0.950 2. Conclusion The process conditions are stable and feasible. The anti-Caco-2 tumor cells in vitro have clear pharmacodynamics, which lays the foundation for the development of anti-cancer drugs with the main component of total flavone purified from Schizonepeta tenuifolia.