黄桷树叶黄酮的提取纯化及抗BV-2细胞炎性活化的作用研究

Extraction and purification of total flavonoids from leaves of Ficus Lacor and its anti-inflammation activation effects of BV-2 cells

目的探讨黄桷树叶黄酮的提取、纯化方法以及纯化产物对BV-2小胶质细胞脂多糖炎性活化的抑制作用。方法采用乙醇浸提和AB-8大孔树脂对黄桷树叶中的黄酮进行提取和纯化;采用BV-2细胞脂多糖炎性活化模型,通过对细胞活力、细胞形态、一氧化氮和白细胞介素-1β(IL-1β)的检测,研究黄酮纯化物对BV-2细胞炎性活化的抑制作用。结果 50%乙醇提取20min具有最高的黄酮提取效率,可达到8.29%;春季树叶具有最高的黄酮含量;AB-8大孔树脂吸附后,70%乙醇洗脱物具有最高的黄酮纯度,为88.95%;20.0、50.0mg/L黄酮纯化物能明显抑制BV-2细胞在1.0mg/mL脂多糖刺激下产生的炎性活化的形态以及细胞培养液中一氧化氮、IL-1β水平升高。结论黄酮纯化物对BV-2小胶质细胞炎性活化具有良好的抑制作用。

Objective To investigate the extraction and purification method of total flavonoids from the leaves of Ficus Lacor and the inhibiting effect of purified products on LPS inflammatory activation of BV-2 cells. Methods The ethanol extraction and AB-8 macroporous resin were used to extract and purify the total flavonoids in the leaves of Ficus Lacor.The lipopolysaccharide (LPS) inflammation activation model of BV-2 cells was adopted.The inhibiting effect of the flavonoids purification on inflammatory activation of BV-2 cells was investigated by detecting the cellular morphology,cell viability,nitric oxide (NO) and IL-1β. Results 50% ethanol extraction for 20 min had the highest total flavonoids extracting efficiency,which reached 8.29 %,moreover the leaves in spring had the highest total flavonoids content.In the purification of the flavonoids extract,after AB-8 macroporous resin adsorption,70% ethanol eluate had the highest flavonoids purity,which was 88.95%.The flavonoids purification material at the concentration of 20.0 mg/L and 50.0 mg/L could significantly inhibit the inflammatory activation morphology of BV-2 cells under 1.0 mg/mL LPS stimulation and the increase of NO and IL-1β contents in the cell culture fluid. Conclusion Ethanol purification material has good inhibiting effect on the inflammatory activation of BV-2 cells.