Study On The Extraction And Isolation Of Laggerapterodontaand Its Anti-tumor Activity

Laggerapterodonta(DC.)Benth.which showed anticancer activity, is indigenous to China. The plant is used for thetreatment of upper respiratory tract infection, tonsillitis, laryngitis. In order to findactive agents from it, the study on its chemical constituents was carried out.By means of extraction,AB-8macroporous resin column, Sephadex LH-20 gel column, recrystallization, PR-HPLC and so on.5compounds were isolated from the 95% ethanol extract ofLaggerapterodonta (DC.) Benth. Their structures were identified by their physicochemical characteristics and detailed spectroscopic analysis.5 compounds were identified asartemitin, chrysosptertin B,5-Hydroxy-3’,4’,7-trimethoxyflavone, retusineandapigemin-7-O-β-D-glucopyranoside.Four parts are from the 95% ethanol extract through AB-8 macroporous resin column. They are water,50% ethanol,70% ethanol and 95% ethanol elution parts. The four parts were separated by HPLC for their chemical composition analysis,39.96% of 70% ethanol elution part and 84.99% of 95% ethanol elution part are artemitin, chrysosptertin B and retusine.The inhibition effect of the extraction and isolation of Laggerapterodonta on A549 cell, MCF-7 cell, SMMC-7721 cell were assessed by the MTT assay, and the results showed that the anticancer activity of four parts and artemitin, chrysosptertin B had antitumor activity in a dose-dependent manner. Except water and 50% ethanol elution part,70% ethanol and 95% ethanol elution parts have good antitumor activity and artemitin is the best one.Cell corpuscles and fragments of A549 induced by 100μg·ml-1 of 70% and 95% ethanol elution parts were detectedby Hoechst33342 staining assay. Then, the early apoptosis of A549 cells inducing by artemitin and 95% ethanol elution part were detected by the Annexin V and propidium iodide double-staining assay. The results showed that the treatment of them induced apoptosis of A549, and with the dosage increased, apoptosis peak increased.