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目的采用AB-8大孔树脂纯化饿蚂蝗总黄酮,并评价其体外抗HBV活性。方法采用AB-8大孔树脂为吸附材料,对影响纯化效果的各因素进行考察,优选工艺条件。运用Hep G2.2.15细胞模型研究饿蚂蝗总黄酮对细胞的毒性及对HBV抗原分泌的影响。结果 AB-8大孔树脂的最佳纯化工艺为:上柱药液浓度以生药计为0.4 g·m L-1,上样量为1.5 BV(树脂床体积),以4 BV的70%乙醇洗脱,洗脱流速为2 m L·min-1;纯化后总黄酮含量为61.47%。饿蚂蝗总黄酮对Hep G2.2.15细胞的毒性较小,TC50为310.91μg·m L-1;对细胞上清液中HBs Ag和HBe Ag的分泌有明显抑制作用,其对HBs Ag和HBe Ag的IC50分别为56.25μg·m L-1和39.70μg·m L-1。结论 AB-8大孔树脂能较好地用于饿蚂蝗总黄酮的纯化,饿蚂蝗总黄酮在体外有抗HBV作用。
OBJECTIVE To purify total flavonoids of hungry locust by AB-8 macroporous resin and evaluate the anti-HBV activity in vitro. Methods AB-8 macroporous resin was used as adsorbent to investigate the factors influencing the purification and optimize the process conditions. The Hep G2.2.15 Cell Model was Used to Study the Toxicity of Total Flavonoids from Locusta migratoriae and Their Effects on HBV Antigen Secretion. Results The optimum purification process of AB-8 macroporous resin was as follows: the concentration of the liquid on the column was 0.4 g · m L -1 as crude drug, the sample volume was 1.5 BV (resin bed volume) The elution flow rate was 2 m L · min-1. The content of total flavonoids after purification was 61.47%. The total flavonoids of the hungry locust were less toxic to Hep G2.2.15 cells with a TC50 of 310.91μg · m L-1, which significantly inhibited the secretion of HBsAg and HBeAg in the supernatant, The IC50 of Ag was 56.25μg · m L-1 and 39.70μg · m L-1, respectively. Conclusion The AB-8 macroporous resin can be used to purify the total flavonoids of the hungry locust. The flavonoids from the hungry locust have anti-HBV effect in vitro.