Study On Differentiation-Inducers Against K562 From Fructus Arctii

Fructus Arctii is the dry mature fruit of Arctium lappa L.which is a perennial herb from Compositae. In this thesis, chemical constituents from Fructus Arctii are isolated, their structures are identified, and the activity of inducing Leukemia cell K562 are evaluated, the result are as follow:1. Separation and purification of chemical component in Fructus ArctiiSilica gel column chromatography was used to separate the extract of Fructus Arctii preliminary, ethyl acetate-methanol or methylene chloride-methanol was selected with gradient elution according to a certain ratio, the separative products was purified by high performance liquid chromatography (HPLC), a method was established both under isocratic and gradient elutions with methanol-water or acetonitrile-water. Result: Compound I was obtained in the preliminary separation of the extracts. The preliminary extracts are purified with HPLC, and compound Ⅱ～Ⅶ are acquired. In this separation, eight compounds are obtained in total, with analysis of HPLC, the requirement of structure measurement are contented.2.Structure identification of Fructus Arctii chemical component.UV, IR, ESI-MS, NMR were used to parse the chemical structure of the eight compounds in the separation and purification. With all the data and report, the structures of the eight compounds are confirmed, compound Ⅰ～Ⅶare arctiin, lappaol F, matairesinol, lappaol C, arctignan B, lappaol E, lappaol A, isomer of lappaol A. 3.Evaluation of induced differentiation against K562 in vitro by Fructus Arctii effective ingredientsTrypan blue staining method was used to test the survival rate of K562 in different drug concentration at different time. Cell cycle and expression of cell-surface differentiation antigen CDllb were measured by flow cytometry (FCM). We researched differentiation-inducing activity of K562 with lappaol F, matairesinol and lappaol C. It suggested that there is no cytotoxicity for K562 in the concentration from 20μmol/L to 100μmoI/L. K562 cells are stoped in period G0/G1, the minimum inhibitory concentration(MIC) is 20μmol/L, It is detected that the positive expressing of differentiation antigen CD11b, which on the surface of K560, increasing with drug treatment. In conclusion, lappaol F, matairesinol and lappaol C all exert differentiation-inducing activity to a certain extent.