卷柏总黄酮的纯化工艺与细胞保护作用

Purification Process of Total Flavonoids from Selaginella pulvinata and Protecting Effects on Neuroblastoma Cells

卷柏(Selaginella tamariscina)含有丰富的黄酮类成分,以双黄酮为主。利用AB-8大孔树脂对脂溶性成分的吸附作用和聚酰胺树脂对黄酮类成分吸附和解吸的特点,将二者串联对卷柏提取物进行分离纯化,建立垫状卷柏总黄酮(Total flavonoids of selaginella pulvinata,TFSP)提取纯化工艺。采用紫外分光光度法测定TFSP含量,并通过MTT法,检测TFSP对神经细胞过氧化氢损伤的保护与修复作用。结果表明,垫状卷柏提取物先通过AB-8大孔树脂吸附脂溶性成分,再经聚酰胺吸附和解吸卷柏总黄酮,在聚酰胺树脂上样量1∶2(卷柏与树脂的体积比)、80%乙醇洗脱5∶1(洗脱液与聚酰胺柱的体积比)等最佳纯化工艺条件下,TFSP的平均含量达81.4%,工艺过程科学合理、简便易行。将TFSP与SH-SY5Y细胞共孵育,可显著减轻神经细胞被过氧化氢损伤的程度,并对已经受损的细胞具有明显的修复作用。TFSP具有为神经退行性疾病开发治疗药物的潜力。

s:Flavonoids,especially biflavonoids,are main components of Selaginella tamariscina and Selaginella pulvinate.The purification process of total flavonoids from Selaginella pulvinata(TFSP)was established by purifying the Selaginella pulvinata extract through AB-8 macroporous resin followed by polyamide.The content of TFSP was determined by ultravio⁃let spectrophotometry.The protecting and repairing effects of TFSP on hydrogen peroxide induced neuron cell damage were detected by MTT.The results showed that AB-8 macroporous resin adsorbed and removed the liposoluble substance from the extract,and flavonoids were isolated through adsorption-desorption in polyamide column.The content of TFSP yielded from the process was up to 81.4%under the optimal conditions—polyamide resin uploading volume of 1∶2(C.matta vs.resin),and 80%ethanol eluent of 5∶1(eluent vs.polyamide column).The purification process is scientific,reasonable,simple and easy to implement.TFSP showed remarkable activity on neuroblastoma cell and increased the cell viability before or after it was cocultured with SH⁃SY5Y cells.Therefore,TFSP has good effects on nerve cell peroxidation damage with the potential to be developed as therapeutic drug for neurodegenerative disorder.