拟草果总黄酮制备纯化及抗炎活性研究

Purification and anti-inflammatory activity of total flavonoids from Amomum paratsao-ko

关键词:  拟草果, 总黄酮, 大孔吸附树脂, 纯化工艺, 抗炎活性

Key words:  Amomum paratsao-ko, total flavonoids, macroporous adsorption resin, purification process, anti-inflammatory activity

摘要:

该文在前期研究的基础上,以拟草果的甲醇粗提液为原料,研究了拟草果总黄酮成分的纯化方法以及考察拟草果总黄酮的抗炎活性。结果表明:采用静态吸附-洗脱试验筛选HP-20为纯化拟草果总黄酮的最佳大孔吸附树脂; 以吸附率、解吸率等参数为指标,考察上样液和洗脱液的质量浓度、体积、流速等因素对纯化工艺的影响,确定最佳工艺条件:上样液质量浓度为0.5 mg·mL^-1、上样体积流量为4 mL·min^-1、上样体积为15 BV、洗脱剂乙醇浓度为70%、洗脱流速为2 mL·min^-1、洗脱剂用量为10 BV,在此条件下纯化的总黄酮保留率为65.48%; 通过检测获得的拟草果总黄酮对脂多糖刺激的小胶质BV2细胞中白介素(IL)-6水平的影响发现,其可显著下调炎症因子IL-6的表达,具有一定的抗炎活性。

Abstract:

This article aimed to develop a separation and purification method for the total flavonoids of the methanol extraction from Amomum paratsao-ko, and the anti-inflammatory activity of total flavonoids from A. paratsao-ko was also investigated. The static adsorption-elution test screened out the HP-20 as the best macroporous adsorption resin for purifying the total flavonoids of A. paratsao-ko. The comprehensive scores of adsorption and desorption rates were used as the indexes to determine the optimal purification process parameters as follows: the concentration of the upper column sample solution 0.5 mg·mL^-1, the sample flow rate 4 mL·min^-1, and the upper column volume 15 BV, 70% ethanol 10 BV for elution, the elution flow rate 2 mL·min^-1. The obtained total flavonoids of A. paratsao-ko were evaluated for the anti-inflammatory activity via examination of the effect on IL-6 in LPS-stimulated BV2 microglial cells. The results showed that the total flavonoids of A. paratsao-ko significantly inhibit IL-6 expression in the LPS-stimulated BV2 microglial cells.